Neuroscience and Biobehavioral Reviews 55 (2015) 119–146

Contents lists available at ScienceDirect

Neuroscience and Biobehavioral Reviews

journal homepage: www.elsevier.com/locate/neubiorev

Review

Long-term consequences of perinatal and adolescent cannabinoid

exposure on neural and psychological processes
Alejandro Higuera-Matas ∗ , Marcos Ucha, Emilio Ambrosio
Department of Psychobiology, School of Psychology, National University of Distance Learning (UNED), C/ Juan del Rosal 10, 28040 Madrid, Spain

a r t i c l e i n f o a b s t r a c t

Article history: Marihuana is the most widely consumed illicit drug, even among adolescents and pregnant women. Given
Received 29 October 2014 the critical developmental processes that occur in the adolescent and fetal nervous system, marihuana
Received in revised form 30 March 2015 consumption during these stages may have permanent consequences on several brain functions in later
Accepted 29 April 2015
adult life. Here, we review what is currently known about the long-term consequences of perinatal and
Available online 8 May 2015
adolescent cannabinoid exposure. The most consistent findings point to long-term impairments in cogni-
tive function that are associated with structural alterations and disturbed synaptic plasticity. In addition,
Keywords:
several neurochemical modifications are also evident after prenatal or adolescent cannabinoid exposure,
Cannabinoid
Cognition
especially in the endocannabinoid, glutamatergic, dopaminergic and opioidergic systems. Important sex-
Emotion ual dimorphisms are also evident in terms of the long-lasting effects of cannabinoid consumption during
Drug addiction pregnancy and adolescence, and cannabinoids possibly have a protective effect in adolescents who have
Prenatal development suffered traumatic life challenges, such as maternal separation or intense stress. Finally, we suggest some
Adolescence future research directions that may encourage further advances in this exciting field.
Dopamine © 2015 Elsevier Ltd. All rights reserved.
Glutamate

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
2. Epidemiology of marihuana use during critical developmental periods: pregnancy and adolescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
3. The endocannabinoid system during perinatal development and adolescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
4. Role of the endocannabinoid system in the regulation of central nervous system development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
5. Perinatal cannabinoid exposure and long-lasting effects on neural and psychological processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
5.1. Human studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
5.1.1. Cohort studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
5.1.2. Neuroimaging studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
5.2. Animal studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
5.2.1. Learning and memory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
5.2.2. Emotional regulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
5.2.3. Sensitivity to drugs of abuse later in life and potential addictive behavior. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
5.2.4. Monoaminergic systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
5.2.5. Glutamatergic and GABAergic systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
5.2.6. Endogenous opioid system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
5.2.7. Endogenous cannabinoid system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
6. Adolescent cannabinoid exposure and long-lasting effects on neural and psychological processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
6.1. Human studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
6.1.1. Neuropsychological assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
6.1.2. Neuroimaging studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129

∗ Corresponding author at: Departamento de Psicobiología, Facultad de Psicología, UNED, C/ Juan del Rosal 10, 28040 Madrid, Spain. Tel.: +34 913989689;
fax: +34 913987491.
E-mail address: ahiguera@psi.uned.es (A. Higuera-Matas).

http://dx.doi.org/10.1016/j.neubiorev.2015.04.020
0149-7634/© 2015 Elsevier Ltd. All rights reserved.
120 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146

6.2. Animal studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129

6.2.1. Learning and memory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
6.2.2. Emotional regulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
6.2.3. Sensitivity to drugs of abuse later in life and potential addictive behavior. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
6.2.4. Monoaminergic systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
6.2.5. Glutamatergic and GABAergic systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
6.2.6. Endogenous opioid system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
6.2.7. Endogenous cannabinoid system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
6.2.8. Cholinergic system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
6.2.9. Other signaling molecules and brain metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
6.2.10. Structural and synaptic plasticity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
6.2.11. Cannabinoid exposure during adolescence as a normalizing agent against the deleterious effects of life challenges . . . . . . . . . 139
7. Concluding remarks and future directions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140

1. Introduction contributions into the long-lasting consequences of cannabinoid

exposure during adolescence. As for prenatal intake, there are also
Cannabis remains the most widely used illicit substance world- excellent reviews on cannabinoid exposure during adolescence and
wide, with an estimated annual prevalence of 3.9% of the adult the ensuing adult behavioral alterations (Chadwick et al., 2013;
population in 2013 (around 180 million users aged 15–64 years), Hurd et al., 2014; Realini et al., 2009; Renard et al., 2014; Rubino and
yet with marked regional differences (1.9% prevalence in Asia and Parolaro, 2008; Schneider, 2009, 2008; Trezza et al., 2012; Viveros
10.9% in Oceania for example: United Nations Office on Drugs and et al., 2011a,b), leading us to focus mainly on the most consistent or
Crime, 2013). The debate surrounding the possible legalization of interesting findings. We will first review the studies on long-term
cannabis has become quite lively, and with a growing number of cognitive alterations induced by adolescent cannabinoid exposure
countries and states in the USA having taken this step or contem- (see Table 5), before focusing on the emotional changes (including
plating doing so, it is crucial that such a decision is made based on anxiety or depressive-like behavior: Table 6), thereafter assessing
firm scientific grounds. the data available on the altered sensitivity to drugs of abuse found
Prenatal stages and adolescence are two crucial periods when in cannabinoid-exposed individuals. Subsequently, we will review
exposure to drugs of abuse affects the ongoing development of the the most important alterations found in the main neurotransmit-
central nervous system (CNS), altering the normal psychological ter systems (see Table 7), as well as focusing on the alterations
function of the individual. In this review, we will mainly focus on in morphological and synaptic plasticity induced by adolescent
the long-term effects of exposure to “cannabinoid agents” during cannabinoid exposure, and on some of the possible underlying
pregnancy and adolescence (see Table 1 for a list of the cannabinoid molecular mechanisms. Special attention will be paid to possible
agents cited in this review and their binding properties), while we protective actions of early cannabinoid exposure after traumatic
will not deal with acute effects of cannabinoids or those alterations life events, such as chronic stress or maternal deprivation.
studied immediately after chronic or repeated exposure. Sexual We believe that updating our knowledge of the long-term effects
dimorphism in the effects reported will be highlighted whenever of cannabis exposure during these two periods should help to clar-
studies were carried out on both sexes. Unless otherwise specified, ify how to cut the Gordian knot of marihuana legalization.
the sex of the animals tested is always masculine.
Although human studies provide the most relevant informa- 2. Epidemiology of marihuana use during critical
tion regarding the long-term influences of prenatal marihuana developmental periods: pregnancy and adolescence
exposure on later cognitive development and neural plasticity,
several confounding factors might limit the conclusions that can Marihuana is the illicit drug most widely consumed during preg-
be obtained from such studies, such as a possible malnutrition nancy (Brown and Graves, 2013; Calvigioni et al., 2014; Jaques
derived from the maternal consumption of cannabis, different 9 - et al., 2014), its consumption being strongly associated to cigarette
tetrahydrocannabinol (THC) consumption over time and between smoking (Gaalema et al., 2013) and to a lesser extent, to opi-
studies, or differences in educational background or socioeconomic oids, stimulants and alcohol (Burns et al., 2006). In the United
status. For this reason, animal studies are an invaluable tool when it States, approximately 4.6% of pregnant women reported having
comes to draw causal conclusions regarding the long-term effects consumed marihuana in the past month during the first trimester
of perinatal cannabinoid exposure. There are excellent reviews of pregnancy. This percentage went down to 2.9% during the second
on the behavioral consequences of prenatal THC and cannabinoid trimester and subsequently decreased to 1.4% in the third trimester
intake during pregnancy and lactation, summarizing our current (Substance Abuse and Mental Health Services Administration,
understanding of this field (Calvigioni et al., 2014; Campolongo 2009). In Brazil this percentage is similar, around 4.0% in the first
et al., 2011; Morris et al., 2011; Trezza et al., 2012; Wu et al., trimester (Mitsuhiro et al., 2007), as is the prevalence of its con-
2011). As such, we will only briefly review their most significant sumption in the general population of pregnant women in Europe.
findings. Along with these behavioral effects, prenatal exposure to For example, the prevalence of marihuana use during pregnancy
cannabinoids has consistently been associated with dysregulated in the UK was reported to be around 5% (Fergusson et al., 2002),
neurotransmission in the brain. Thus, we will also describe what is with a similar prevalence in Spain, where THC was detected in 5.3%
currently known about the neurochemical alterations induced by of newborns by meconium analysis (Lozano et al., 2007: although
in utero cannabis (see Table 4). this percentage rises to 10.3% in Ibiza, in the Balearic Islands
As for prenatal studies, human cohort designs of adolescent Friguls et al., 2012). In France, 1.2% of women reported having used
cannabis users and abusers might again include confounding fac- cannabis during pregnancy, a percentage that rose among younger
tors that affect the reliability of the results obtained. For this women, women living alone, or women who had a low level of
reason, the animal studies addressing this issue are crucial research education or low income (Saurel-Cubizolles et al., 2014). Notably,
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
Table 1
Ki values for ligands at human and rat cannabinoid receptors.
Human
CB1
9 -THC 25.1 ± 5.54
8 -THC NA
Cannabinol 525.3 ± 308.1
Cannabidiol 27542
CP 55,940 3.1 ± 1.31
WIN 55,512-2 62.4 ± 34.96
Anandamide 239.2 ± 61.77
2-Arachidonoylglycerol 3423.6 ± 3288.24
Adapted from Howlett et al. (2002), McPartland et al. (2007), Pertwee (2008).
when compared to the year prior to the survey, the reduction in
the prevalence of marihuana use during pregnancy in the UK is less
than in the United States (see Moore et al., 2010), which is par-
ticularly interesting given that adolescent mothers are particularly
vulnerable and at risk for increased substance use when compared
with their nulliparous peers (reviewed in Chapman and Wu, 2013).
Due to their young age, these mothers are likely to have subsequent
pregnancies that could also be adversely affected by substance use
(Cornelius et al., 2004). Indeed, even if the biological consequences
of prenatal substance use are disregarded, postpartum use is a risk
factor for substance abuse in children (U.S. Department of Health
and Human Services, 1999).
Marihuana is also the illicit drug most widely consumed dur-
ing adolescence both in the United States (Substance Abuse
and Mental Health Services Administration, 2014a) and Europe
(European Monitoring Centre for Drugs and Drug Addiction, 2014).
In 2013, male adolescents showed a higher prevalence of last
month use (approximately 8.4%) than female adolescents (approx-
imately 5.6%) in the United States (Substance Abuse and Mental
Health Services Administration, 2014b). In Europe this same trend
is also observed with around 60% of the adolescents who reported
cannabis use in the last month being men (European Monitoring
Centre for Drugs and Drug Addiction, 2014). In Spain, cannabis is
also the illicit drug most widely consumed by adolescents (around
26.6% of people in this age segment report cannabis consumption in
the last month), where male adolescents again show higher preva-
lence rates than females (Delegación del Gobierno para el Plan
Nacional sobre Drogas, 2013).
In the light of these data, it is crucial to understand what are the
potential long-term consequences of cannabis exposure at these
crucial stages of development. In this review, we shall first outline
the stages of nervous system development at which endogenous
cannabinoids (eCBs) play a major role. This will help us to under-
stand how cannabis exposure can alter the normal developmental
trajectories in the nervous system, thereby perturbing the neu-
ral plasticity that constitutes the basis of cognitive processes and
motivation.
3. The endocannabinoid system during perinatal
development and adolescence
The endocannabinoid (eCB) system is present from the earli-
est phases of development (Maccarrone et al., 2014). Indeed, in
rodents it has been reported that the CB1 receptor (CB1R) is present
and functionally active in the preimplantation embryo and in the
uterus (Paria and Dey, 2000). Furthermore, eCB signaling at these
stages is intimately associated with embryo–uterine interactions
during implantation. The deletion of the CB1R compromises normal
cell proliferation and differentiation, leading to aberrant placenta-
tion and compromising embryo implantation (Paria and Dey, 2000;
Sun and Dey, 2008). Like the CB1R, the CB2 receptor (CB2R) is also
expressed in mouse embryos from the blastocyst stage and it may
121
Rat
CB2 CB1 CB2
35.2 ± 5.86 42.6 ± 5.01 13 ± 7.7
NA 47.6 39.3
168.2 ± 32.16 368 ± 121.1 126.4
2629.5 ± 230.5 2210.5 ± 558.08 1000
1.68 ± 0.54 0.94 ± 0.44 2.09 ± 0.72
2.56 ± 1.16 7.17 ± 2.77 8.7 ± 7.5
168.2 ± 32.16 368 ± 121.1 279
1193.8 ± 327.71 1180.5 ± 538.59 1900 ± 1800
play a role in embryonic stem-cell derived haematopoietic cell pro-
liferation and lineage differentiation (Jiang et al., 2007).
During brain development, the CB1R is expressed from very
early stages and its distribution within the brain resembles that
found in adults, as seen in rats (Berrendero et al., 1999, 1998;
Rodriguez de Fonseca et al., 1993) and humans (Glass et al., 1997;
Mato et al., 2003; Wang et al., 2003; Zurolo et al., 2010). The expres-
sion of the receptor in both species gradually increases during
development and it is found in areas related to the control of move-
ment (e.g. the basal ganglia, cerebellum), cognition and attention
(e.g. the cerebral cortex), as well as emotions and memory (e.g. the
amygdala, hippocampus). Surprisingly, the CB1R is also present in
the white matter of the rat developing brain, where the receptor
is not expressed significantly in the adult (Romero et al., 1997).
This phenomenon has also been seen in humans (Mato et al., 2003),
where CB1R expression is elevated in the globus pallidus (Glass et al.,
1997; Wang et al., 2003) and substantia nigra pars reticulata (Glass
et al., 1997) when compared to the adult brain. During human
corticogenisis, the CB1R is expressed by Cajal-Retzius cells of the
marginal zone, by migrating post-mitotic neurons, and by apical
and basal progenitors in the proliferative area of the ventricular
and subventricular zones (Saez et al., 2014; Zurolo et al., 2010). The
expression of CB1R by projection neurons increases gradually as
differentiation proceeds, reaching a peak at the moment they reach
the cortical plate (Zurolo et al., 2010). All these findings suggest a
specific role of the eCB system in brain development.
Although CB1R mRNA expression in the rat brain does not
change from the postnatal period to adulthood (McLaughlin et al.,
1994), CB1R binding gradually increases in neonates, reaching a
peak around the pubertal period before decreasing to the adult lev-
els (Belue et al., 1995; McLaughlin et al., 1994; Rodriguez de Fonseca
et al., 1993). There are some sexual dimorphisms in the develop-
ment expression of the CB1R. For example, subtle sex differences
in the binding to CB1R were detected in the rat striatum and ven-
tral mesencephalon but not in the limbic forebrain (Rodriguez de
Fonseca et al., 1993). Similarly, neonatal male rats displayed signifi-
cantly higher levels of CB1Rs in CA3 while they had less CB2R in the
polymorphic layer of the dentate gyrus (DG) (Suarez et al., 2009).
In adolescence, subtle differences in the expression of hippocam-
pal CB1Rs were found with a lower density of CB1Rs in female rats
than in males (Marco et al., 2007). In humans, the CB1R increased
from the postnatal period to adulthood (Mato et al., 2003), although
CB1R mRNA expression peaks in neonates and toddlers, there-
after decreasing gradually over the individual’s lifespan (Long et al.,
2012).
Regarding the two major eCBs, 2-arachidonoylglycerol (2-AG)
concentrations during brain development (2–8 nmol/g tissue)
exceed those of N-arachidonoylethanolamine (Anandamide or
AEA) (3–6 pmol/g tissue) in whole rat brain lysates (Berrendero
et al., 1999), the latter being detected at low concentrations at
mid-gestation. AEA levels rise gradually during the perinatal
period in the rat brain, reaching a peak during adolescence before
122 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
decreasing to the previous levels in most of the brain areas studied
up to date (Berrendero et al., 1999; Ellgren et al., 2008; Lee et al.,
2013; Rubino et al., 2014; Wenger et al., 2002). After that, AEA
levels continue to rise until they reach adult levels (Lee et al.,
2013). Concomitantly, the AEA synthetic and hydrolytic enzymes,
N-acyl phosphatidylethanolamine phospholipase D (NAPE-PLD)
and fatty acid amide hydrolase (FAAH), augment in the whole brain
as postnatal life progresses (Morishita et al., 2005), something
also seen in the human dorsolateral prefrontal cortex (PFC) (Long
et al., 2012). 2-AG concentrations also increase in mice and rats
during development but by contrast, there is a peak in cortical and
hippocampal 2-AG concentrations around the moment of birth
(Berrendero et al., 1999; Keimpema et al., 2010). This is consistent
with an increase in diacylglycerol lipase (DAGL) expression that
begins on GD14.5/GD16.5 (GD: Gestational Day) and that remains
high thereafter until the moment of birth, when it decreases
abruptly (Keimpema et al., 2010). In the rat PFC and nucleus
accumbens (NAcc) 2-AG levels were seen to be lower at later
stages than at the beginning of adolescence (Ellgren et al., 2008). In
humans, DAGL␣ mRNA is more strongly expressed in this period
(between school age and young adulthood) than at any other stage,
and monoacylglycerol lipase (MAGL) expression decreases after
peaking in infancy (Long et al., 2012).
These differences in the expression of several elements of the
eCB system during gestation and adolescence suggest a role for this
system in the changes the brain undergoes during those stages,
reflecting periods of high vulnerability for the consequences of
exogenous cannabinoid administration.
4. Role of the endocannabinoid system in the regulation of
central nervous system development
The question arises as to the role the eCB system fulfills dur-
ing CNS development. Since the seminal work showing that eCBs
regulate interneuron migration and morphogenesis by transacti-
vating the TrkB receptor (Berghuis et al., 2005), considerable effort
has focused on the influence of eCB signaling on neuronal develop-
ment. It was shown that eCBs trigger CB1R internalization and its
elimination from filopodia, and they induce chemorepulsion and
the collapse of GABAergic (GABA: ␥-aminobutyric acid) interneu-
ron axonal growth cones (Berghuis et al., 2007). In addition, how the
subcellular distribution of MAGL in developing neurons establishes
2-AG microgradients that modulate both growth cone matura-
tion and axonal elongation was described (Keimpema et al., 2010).
Moreover, the target selection of GABAergic interneurons lacking
the CB1R was shown to be impaired, highlighting the function of
eCBs as axon guidance cues (Berghuis et al., 2007). Signaling at CB1
and CB2 receptors also influenced neural progenitor proliferation
(Aguado et al., 2005; Mulder et al., 2008) in part by its action on the
mTORC1/Pax6 pathway that drives Trb2 expression and basal pro-
genitor expansion (Díaz-Alonso et al., 2014). In the subventricular
zone, intermediate progenitor cells that contribute to the gener-
ation of pyramidal cells are targeted by CB1R, suggesting a role
of the eCB system in cell fate regulation (Kowalczyk et al., 2009).
Moreover, eCB signaling controls the radial migration of pyrami-
dal cells, and the elongation and fasciculation of their long-range
axons once they settle in a cortical layer (Mulder et al., 2008), such
as that of the corticothalamic projections (Wu et al., 2010). How-
ever, the pharmacological elevation of AEA through FAAH inhibitors
during mid-gestation in mice did not affect neurogenesis or axonal
development when measured at GD16.5 (Wu et al., 2014). One
of the main effectors of eCB’s influence on cortical development
is the Slit2/Robo1 signaling pathway, with a critical involvement
of CB1R activation (Alpár et al., 2014). CB1Rs also influence the
migration of other cells, like that of neuroblasts along the rostral
migratory stream (Oudin et al., 2011) and interneurons during cor-
ticogenesis (Berghuis et al., 2005). eCB signaling participates in the
development of other neurotransmitter pathways, as seen in the
cholinergic system where 2-AG signaling promotes the differenti-
ation and synaptic connectivity of cholinergic projection neurons
(Keimpema et al., 2013). The development of glutamatergic neu-
rons might also be regulated by the eCB system. Indeed, prenatal
exposure to the CB1 and CB2 receptor agonist WIN 55,212-2 (WIN)
alters migration of early-born glutamatergic neurons and GABAer-
gic interneurons in the rat cerebral cortex, an effect that might
depend on the transcription factors Tbr1 and Tbr2 (Saez et al., 2014).
Correct axon guidance in the retinothalamic pathway is another
developmental process that requires the presence of CB2Rs (Duff
et al., 2013), suggesting that both cannabinoid receptors are impor-
tant in shaping the developing nervous system in mammals. The
eCB system is also thought to fulfill an important role in synapto-
genesis (Kim and Thayer, 2001) and in the modulation of synaptic
transmission in the developing hippocampus (Zhu and Lovinger,
2010). Taking into consideration all the data presented above, it is
not surprising to find that repeated THC exposure disrupts eCB sig-
naling, particularly the temporal dynamics of the CB1R in rewiring
the fetal cortical circuitry. This phenomenon appears to have the
Superior Cervical Ganglion 10 (SCG10)/stathmin-2 pathway as a
substrate and it has been identified as the first cannabis-driven
molecular effector in the developing cerebrum (Tortoriello et al.,
2014).
In the light of these findings, the consequences of in utero THC
exposure on the development of the CNS in humans still remain
unclear. To our knowledge there is only one collection of human
fetal specimens with documented marihuana exposure (DiNieri
et al., 2011; Hurd et al., 2005; Wang et al., 2006, 2004). When
studied, early marihuana exposure was shown to impair growth in
mid-gestation fetuses and to diminish the mRNA expression of the
dopaminergic receptor D2 in the amygdala and NAcc (no changes
were observed in the dorsal striatum: DiNieri et al., 2011; Hurd
et al., 2005; Wang et al., 2004). Notably, in the amygdala, the differ-
ences reported were only found in male fetuses (Wang et al., 2006).
In addition, it was also shown that prenatal marihuana exposure
decreases the levels of preproenkephalin (PENK) gene expression
in the fetal striatum (Wang et al., 2006), while the levels of prepro-
dynorphin (PDYN) transcripts remained unaffected (DiNieri et al.,
2011; Wang et al., 2006). Opioid receptors were also affected in
the fetal brain by in utero cannabis exposure, with increased ␮ opi-
oid receptors in the amygdale and decreased levels of the ␬ opioid
receptors in the mediodorsal thalamus (Wang et al., 2006). Lastly,
as mentioned before, maternal cannabis smoking was associated
to reduced SCG10/stathmin-2 mRNA and protein expression in this
collection, a fact that was proposed to be a key molecular effector
mediating adverse effects of cannabis (Tortoriello et al., 2014).
5. Perinatal cannabinoid exposure and long-lasting effects
on neural and psychological processes
5.1. Human studies
5.1.1. Cohort studies
The issue of the long-term consequences of prenatal cannabis
exposure has been widely studied in three human cohorts: the
Ottawa Prenatal Prospective Study (OPPS), the Maternal Health
Practices and Childhood Development Project (MHPCD) and more
recently, the Generation R study. While the results of these stud-
ies are reviewed elsewhere (Calvigioni et al., 2014; Huizink, 2014;
Jaques et al., 2014; Morris et al., 2011; Wu et al., 2011), let
us just mention the most important findings. First, in both the
OPPS and MHPCD studies the use of cannabis during pregnancy
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
was not associated with increased miscarriage rates, premature
deliveries or any other complications. In the Generation R study,
another cohort was also studied in depth (El Marroun et al., 2010,
2011a,b, 2012; El Marroun et al., 2008, 2009), showing that mater-
nal cannabis use during pregnancy was associated with growth
restriction in mid and late pregnancy, and with lower birth weight.
Since no such association was found for paternal cannabis use in
the same period, this would appear to represent a direct biological
effect of maternal intrauterine exposure to cannabis on fetal growth
(El Marroun et al., 2010). It is of more concern that the data from
these OPPS and MHPCD cohorts suggest that newborns already dis-
play cannabis-induced neurobehavioral alterations (<1 week old),
as do children later in development. The essential findings from
these cohorts indicate that prenatal marihuana exposure increases
tremor and enhances the startle response in the newborn, and it
is also associated with a wide variety of cognitive deficits (such
as impaired sustained attention, self-control deficits and impaired
executive function), as well as increased aggression and inattention
(in this case only in girls: Huizink, 2014). Moreover, in a different set
of studies, prenatal cannabis exposure was also associated with an
increased risk of nicotine addiction and marihuana use in adulthood
(Day et al., 1994, 1993, 2006, 2011). Some of the aforementioned
findings were confirmed in a smaller study (Noland et al., 2005),
suggesting that the severity of marihuana exposure during preg-
nancy was positively correlated with impaired sustained attention
in pre-school children.
5.1.2. Neuroimaging studies
Neuroimaging data has been examined to assess the long-term
structural alterations in the white and gray matter associated with
intrauterine marihuana exposure (Derauf et al., 2009). Although
studies are scarce, there is evidence of structural alterations in sub-
jects prenatally exposed to marihuana. For example, a volumetric
magnetic resonance imaging (MRI) study of 10–14 year old children
found a reduced cortical gray matter after intrauterine marihuana
exposure, although the model used was not adjusted for age, gen-
der and other drug exposures (Rivkin et al., 2008). In another study
in which frontal white matter development was assessed using dif-
fusion tensor imaging in children with prenatal cocaine exposure,
intrauterine exposure to both cocaine and marihuana resulted in
higher (worse) mean diffusivity than exposure to cocaine alone
(Warner et al., 2006).
Two other functional MRI (fMRI) studies investigated the
relationship between prenatal marihuana exposure and adoles-
cent/young adult neural functioning in tasks requiring cognitive
effort. In the first, subjects with and without prenatal mari-
huana exposure underwent fMRI while performing a two condition
Go/No-Go task. Although there were no significant group differ-
ences for reaction time and errors of omission for either condition,
the group exposed prenatally made more errors in the more diffi-
cult task. The extent of prenatal marihuana exposure was positively
correlated to bilateral PFC and right premotor cortex activity, while
left cerebellar activity was negatively correlated to the degree
of marihuana exposure. This was interpreted as increased effort
needed to perform the task (Smith et al., 2004). In the second study,
fMRI scans were performed on the same subjects during a visuospa-
tial 2-back test. Although no significant differences in performance
were observed between the prenatally exposed and unexposed
subjects, the amount of prenatal marihuana exposure was signifi-
cantly and positively related to increased activity in the left inferior
and middle frontal gyri, the left parahippocampal gyrus, left mid-
dle occipital gyrus and left cerebellum, and it was significantly and
negatively related to activity in the right inferior and middle frontal
gyri. It was suggested that prenatal marihuana exposure may have
altered the normal lateralization and connectivity of multiple brain
123
sites, which, together with the PFC, are important for performing
complex cognitive tasks (Smith et al., 2006).
5.2. Animal studies
5.2.1. Learning and memory
In a wide variety of tests, several cognitive deficits have consis-
tently been reported in rats exposed to THC or other cannabinoid
agents in utero (see Table 2). For example, a disruption of memory
retention was found in a passive avoidance task in rats exposed
to WIN in utero (Mereu et al., 2003), while learning deficits in
the Morris water maze were detected in juvenile rats that had
been exposed to a cannabis resin during prenatal development
(Kawash et al., 1980). With regard to long-term memory, THC-
exposed rats performed worse than controls in an avoidance test
and they also showed impaired short-term memory in an olfac-
tory memory task (Campolongo et al., 2007). In addition, there
is also evidence of impaired learning in an active avoidance task
in WIN-exposed male rats (Antonelli et al., 2005). More recently,
cannabinoid exposed animals showed no acquisition deficits in a
passive avoidance test, while they exhibited deficits in the con-
solidation process during retention testing (Silva et al., 2012). No
treatment-related effects on acquisition were evident in the active
place avoidance task performed on rats, although a significant treat-
ment effect was observed in reversal performance in males but not
in females. Finally, in an attention task, a smaller percentage of
THC-exposed rats completed the test and they required more trials
to complete it (Silva et al., 2012). A recent study identified elevated
AEA levels during perinatal development in mice due to FAAH inhi-
bition, and working memory performance in the offspring of these
mice was impaired in the T-maze (Wu et al., 2014). WIN expo-
sure during prenatal development was seen failed to alter prepulse
inhibition of the startle response, or the startle response per se
(Bortolato et al., 2006), ruling out a mediating role of sensory motor
gating in the cognitive alterations summarized above.
5.2.2. Emotional regulation
There are conflicting results as to how prenatal cannabinoid
exposure might affect anxiety-related behaviors (see Table 3).
While in one study prenatal THC exposure was associated with
increased exploratory activity of male but not female offspring
in the elevated plus maze (EPM: Rubio et al., 1995), revealing
decreased anxiety, the opposite trend was also found elsewhere
(Trezza et al., 2008). When using another anxiety index, ultrasonic
vocalizations (USVs) in infant rats, similar conflicting results were
obtained with one report showing a decrease in USVs (Antonelli
et al., 2005) and another showing the contrary (Trezza et al., 2008).
If we examine social interactions, the results are again unclear.
In two different studies, evidence of reduced social interactions
was presented (O’Shea et al., 2006; Trezza et al., 2008), while in
another utero exposed rats displayed increased social interactions
(Newsom and Kelly, 2008). There are no clear reasons for these dis-
crepant results, but it would appear that the different cannabinoids
used are not the cause. Indeed, CP 55,940 (CP) or THC was employed
in the first two studies where decreases in social interaction were
found. Perhaps, the specific tests used to measure social interac-
tion in each experiment might account for the differences in the
results reported in these three studies. In any case, further research
is clearly needed to provide a definite answer to the question of
whether perinatal cannabinoid exposure indeed has a long-lasting
influence on emotional behavior.
With regard to depressive behavior, one study that analyzed the
behavior of rats in the forced swimming test (FST) after prenatal
THC treatment found no differences from control rats (Newsom
and Kelly, 2008). By contrast, chronic elevation of AEA (induced by
the blockade of the FAAH enzyme) increased the immobility in this
124 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146

Table 2
Behavioral and cognitive alterations found in adult animals after perinatal cannabinoid exposure.

Species Sex Treatment Test Test age Results Reference

Drug Dose Age

Rat ♂ CP 55,940 0.15, 0.2 and PND4-PND24 Novel object PND53 Memory O’Shea et al., 2006
0.3 mg/kg (i.p.) recognition impairment
for 7 days each
dose
Rat ♂ WIN 55,512-2 0.5 or 1 mg/kg GD5-GD20 Prepulse inhibition PND40, 60 and 80 No significant Bortolato et al., 2006
(s.c.) effects
Rat ♂ 9 -THC 5 mg/kg (p.o.) GD15-PND9 Social recognition PND80 Social memory Campolongo et al., 2007
impaired
Rat ♂–♀ 9 -THC 0.15 mg/kg (i.v.) GD1-GD21 Attention task PND55 Attention impaired Silva et al., 2012
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 Active place PND80 Impaired spatial Antonelli et al., 2005
avoidance learning and
memory
Rat ♂–♀ 9 -THC 0.15 mg/kg (i.v.) GD1-GD21 PND45-46 Impaired Silva et al., 2012
behavioral
flexibility ♂
Rat ♂–♀ 9 -THC 0.15 mg/kg (i.v.) GD1-GD21 Passive place PND22-23 Learning and Silva et al., 2012
avoidance memory
impairment
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND40 and 80 Memory Mereu et al., 2003
impairment
Rat ♂ 9 -THC 5 mg/kg (p.o.) GD15-PND9 Inhibitory PND81 Memory Campolongo et al., 2007
avoidance impairment
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 Homing PND6-12 Impaired memory Antonelli et al., 2005
Rat ♂ 9 -THC 5 mg/kg (p.o.) GD15-PND9 Olfactory memory PND80 Impaired memory Campolongo et al., 2007
task
Mouse ♂–♀ URB597(↑AEA) 10 mg/kg (i.p.) GD10-PND7 T Maze >PND60 Impaired working Wu et al., 2014
memory

test (Wu et al., 2014). However, the FST is contested as an animal
model of depression as it appears to be rather a test of the efficiency
of potential antidepressant drugs (Petit-Demouliere et al., 2005;
Yan et al., 2010).
5.2.3. Sensitivity to drugs of abuse later in life and potential
addictive behavior
There is consensus in the literature that cannabis exposure dur-
ing prenatal development affects the sensitivity to drugs of abuse
later in life. Interestingly, this effect has been studied mainly for opi-
ates, with very little information regarding other classes of drugs.
For example, tolerance to the analgesic properties of morphine was
reported in male rats perinatally exposed to THC (Vela et al., 1995)
and enhanced conditioned place preference (CPP) was evoked by
a moderate dose of morphine in adult rats of both sexes exposed
to THC during perinatal development (Rubio et al., 1995). These
results have been replicated more recently in male rats (DiNieri
et al., 2011). When we tested whether perinatal exposure to THC

Table 3
Emotional behavior alterations after perinatal cannabinoid exposure.

Species Sex Treatment Test Test age Results Reference

Drug Dose Age

Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND24 Elevated plus maze PND70 Anxiety ↓♂ Rubio et al., 1995
Rat ♂ 9 -THC 2.5 mg/kg (p.o.) GD5-PND9 PND80 No significant effects Trezza et al., 2008
5 mg/kg (p.o.) Anxiety ↑
Mouse ♂–♀ URB597(↑AEA) 10 mg/kg (i.p.) GD10-PND7 >PND60 No significant effects Wu et al., 2014
Rat ♂ CP 55,940 0.15, 0.2 and PND4-PND24 Emergence test PND57 No significant effects O’Shea et al., 2006
0.3 mg/kg (i.p.) 7
days each dose
Rat ♂ 9 -THC 2 mg/kg (twice a GD1-PND10 Forced swimming PND99 No significant effects Newsom and Kelly, 2008
day) test
Mouse ♂–♀ URB597(↑AEA) 10 mg/kg (i.p.) GD10-PND7 >PND60 Depressive-like Wu et al., 2014
behavior ↑
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 Homing PND6-12 Emotional reactivity ↓ Antonelli et al., 2005
Rat ♂ 9 -THC 2 mg/kg (s.c.) twice GD1-PND10 Open field PND93 Anxiety ↑ Newsom and Kelly, 2008
a day
Mouse ♂–♀ URB597(↑AEA) 10 mg/kg (i.p.) GD10-PND7 >PND60 No significant effects Wu et al., 2014
Rat ♂ 9 -THC 2.5 mg/kg (p.o.) GD5-PND9 Social interaction PND35 No significant effects Trezza et al., 2008
5 mg/kg (p.o.) Social anxiety ↑
Rat ♂ CP 55,940 0.15, 0.2 and PND4-PND24 PND55 Social anxiety ↑ O’Shea et al., 2006
0.3 mg/kg (i.p.) 7
days each dose
Rat ♂ 9 -THC 2 mg/kg (twice a GD1-PND10 PND97 Social anxiety ↓ Newsom and Kelly, 2008
day)
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 Ultrasonic PND10 Emotional reactivity ↓ Antonelli et al., 2005
vocalizations
Rat ♂ 9 -THC 2.5 mg/kg (p.o.) GD5-PND9 PND12 No significant effects Trezza et al., 2008
5 mg/kg (p.o.) Emotional reactivity ↑
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146 125

Table 4
Neurochemical alterations found in adult animals after perinatal cannabinoid exposure.

Species Sex Treatment Tests Reference

Drug Dose Age Age Results

Monoaminergic systems
Rat ♂ Hashish extract 20 mg/kg (p.o.) GD5-PND24 PND15, 20, 30, 40, TH mRNA: Ms PND15 ↑ Bonnin et al., 1994
70 PND70 ↓; TH levels: Ms
PND15 ↑
Mouse ♂ 9 -THC 50 mg/kg (p.o.) GD18 PND64-94 No significant alterations Dalterio et al., 1984a
found
CBN DA levels: Brain tissue ↓;
5-HT and 5HIAA levels: No
significant alterations found
CBD DA levels: Brain tissue ↓;
5-HT and 5HIAA levels: No
significant alterations found
Mouse ♂–♀ 9 -THC 50 mg/kg (p.o.) GD18 PND69♂/PND67- DA levels: No significant Dalterio et al., 1984b
77♀ alterations found; 5-HT
levels: HT ♀↓
CBN DA and 5-HT levels: No
significant alterations found
CBD DA and 5-HT levels: No
significant alterations found
Rat ♂ 9 -THC 0.15 mg/kg (i.v.) GD5-PND2 PND2 and 62 DRD2 mRNA and D2 receptor DiNieri et al., 2011
density: NAcc PND2 ↓,
PND62 ↓
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND24 >PND70 Ambulation (after SCH23390 Garcia et al., 1996
challenge): ↓
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND24 >PND70 DOPAC/DA ratio: NAcc ♀↓; Gonzalez et al., 2003
VTA ♀↓
Rat ♂  -THC
9
5 mg/kg (p.o.) GD13-PND7 PND90 SERT binding levels: NSt ↑; Molina-Holgado et al., 1993
aHT ↓; RphN ↓; LC ↓; 5-HIAA:
aHT ↑; SN ↑;
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-GD20/PND1 GD20 and PND1 5-HT levels: Di ♂↓↓–♀↓ Molina-Holgado et al., 1996
Rat ♂–♀ 9 -THC 0.1, 0.5 and GD5-PND24 >PND70 Motor response to Moreno et al., 2003
2 mg/kg (p.o.) quinpirole: Immobility ♂↑;
Stereotypes ↓♂ (2 mg/kg
dose)
Motor response to
Apomorphine: Locomotion
♂↓ (2 mg/kg dose)
Mouse ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND0-2 TH+ cells: No significant Tree et al., 2014
alterations found in any of
the areas studied
Rat ♂–♀ (mixed) 9 -THC 10 mg/kg (p.o.) GD0a -PND20 PND10, 20, 40 and No significant alterations Walters and Carr, 1988
60 found
8 -THC 1 mg/kg (p.o.) TH activity: St ↓ (PND60)
CBD 10 mg/kg (p.o.) TH activity: St↓ PND(60); D2
receptor Kd: St ↑ (PND10), D2
receptor density: St ↑
(PND10 and 20)

Glutamatergic and GABAergic systems

Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND1 Basal and K+ evoked Glu: Cx Antonelli et al., 2005
↓(cc); NMDA receptor
function: Cx ↓(cc)
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND1 Basal and K+ evoked Glu: Cx Antonelli et al., 2005
↓(cc)
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND120-150 GABA+ and GAD 65/67+ cells: Benagiano et al., 2007
CbC ↑
Rat ♂ 9 -THC 5 mg/kg (p.o.) GD5-PND20 PND81 Glu levels: PFC ↓ Campolongo et al., 2007
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND40 Glu levels: FC ↓; Glu uptake: Castaldo et al., 2007
FC ↑; GLT1 levels: FC ↑;
EAAC1: FC ↑;
9 -THC 5 mg/kg (s.c.) GD5-GD20 Glu levels: FC↓
Rat ♂ 9 -THC 5 mg/kg (p.o.) GD15-PND9 PND40 Basal and K+ evoked Glu: HP Castaldo et al., 2010
↓; Glu uptake: HP ↓; GLT1
levels: HP ↓(syn); GLAST
levels:HP ↓(syn)
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND24 >PND70 Ambulation (after baclofen Garcia-Gil et al., 1999
challenge): ↓
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND40 and 80 Basal and K+ evoked Glu Mereu et al., 2003
levels: HP ↓
Rat ♂ WIN 55,512-2 0.5 or 0.1 mg/kg GD5-GD20 PND50 Firing pattern and intrinsic Shabani et al., 2011
(s.c.) electrophysiological activity
of Purkinje cells altered
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 PND50 Firing pattern and intrinsic Shabani et al., 2014
electrophysiological activity
of Purkinje cells altered
126 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146

Table 4 (Continued)

Species Sex Treatment Tests Reference

Drug Dose Age Age Results

Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND20 PND20, 30 and 70 Glutamine synthetase levels: Suarez et al., 2002
CbC PND20 ♂↓; PND30 ↓
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND20 PND20, 30 and 70 GluN1 levels: Cb PND30↓ Suarez et al., 2004a
PND70↓; GluN2/3 levels: Cb
PND20↓ PND30↓ PND70↓
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND20 PND20, 30 and 70 GLAST mRNA: Cb ↓; EAAC1 Suarez et al., 2004b
mRNA: Cb ↓; GLAST levels:
Cb ↓; EAAC1:PND20 and
30♂↓, PND20 ♀↓

Endogenous opioid system

Rat Not determined 9 -THC 5 mg/kg (p.o.) GD5-GD16 GD16 PENK mRNA: CPu ↑; Spt ↑; Th Perez-Rosado et al., 2000
↑; HT ↑, Mb ↑
GD5-GD18 GD18 PENK mRNA: Cx ↑; CPu ↑
♂–♀ GD5-GD21 GD21 PENK mRNA: Cx ♂↓; CPu
♂↓-♀↑; PvHN ♂↓-♀↑; VmHN
♂↓-♀↑
Rat Not determined 9 -THC 5 mg/kg (p.o.) GD5-GD16 GD16 No significant alterations Perez-Rosado et al., 2002
found
GD5-GD18 GD18 POMC mRNA: Arc↓; Cx ↓
♂–♀ GD5-GD21 GD21 POMC mRNA: Arc ♂↓-♀↑; Cx
♂↓-♀↑; HbN ♂↓-♀↑; PDYN
mRNA: Cx♀↑; HP ♀↑; PvHN
♀↑
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND24 >PND70 ␮ opioid receptor density: Vela et al., 1998
CPu ♂↓; frPaMC ♀↑; HP(CA3)
♀↑; LAmy ♀↓; AmyPmCN
♂↓-♀↑; VTA ♀↑; PAG ♀↑
Endogenous cannabinoid system
Rat ♂ WIN 55,512-2 0.5 mg/kg (s.c.) GD5-GD20 >PND80 CB1 receptor function: HP ↑; Castelli et al., 2007
St ↓; AEA levels: St ↑; Limb ↓;
FAAH activity: St ↓; Limb ↑;
NAPE-PLD activity: St ↑;
Limb ↓
Rat ♂–♀ 9 -THC 5 mg/kg (p.o.) GD5-PND24 >PND70 CB1 receptor density: Cx Garcia-Gil et al., 1999
(Superficial layers) ♂↓; Arc
♂↑; CB1 mRNA: HP(CA2)
♂↑; Vertical activity (after
SR141716A challenge): ♀↑

would lead to increased morphine self-administration, this was
only the case when a fixed ratio one schedule of reinforcement was
used, and only in female offspring (Vela et al., 1998). This effect does
not seem to be due to general changes in reward processing as there
were no differences in food-reinforced behavior. No differences
in self-administration were found when similar tests were per-
formed for heroin, although THC exposed animals exhibited a lower
latency to press the active lever for heroin and more importantly,
an increased relapse rate after mild stress conditions or during
extinction tests (Spano et al., 2007). As mentioned previously, most
of these studies focused on opiates and to our knowledge only two
studies examined the effects of perinatal cannabinoid exposure
on psychostimulant effects. In the first, both in male and female
THC-exposed rats displayed a blunted locomotor response to an
amphetamine challenge in adulthood, although females were more
active overall (Silva et al., 2012). Similarly, there was a blunted rein-
forcing action of cocaine (measured in the CPP test) in the offspring
of mice exposed to elevated levels of AEA during gestation (Wu
et al., 2014). Another study focused on alcohol self-administration,
asking whether perinatally THC-exposed rats would self-
administer more alcohol as adults and also, if it would alter gene
expression patterns (Economidou et al., 2007). While THC affected
the expression of 112 genes, similar alcohol self-administration
rates were obtained in THC- and vehicle-exposed animals.
5.2.4. Monoaminergic systems
The effects of perinatal cannabinoid treatments on dopaminer-
gic function are controversial. While early reports suggested that
acute prenatal THC did not modify the concentration of brain bio-
genic amines (Dalterio et al., 1984a,b), it was later shown that
in perinatally THC-exposed females but not males had higher
rates of dopamine synthesis in the limbic forebrain but not in the
striatum (Garcia et al., 1996). In addition, the DOPAC/dopamine
(DOPAC: 3,4-dihydroxyphenylacetic acid) ratio in the NAcc and
ventral tegmental area (VTA) was lower in THC-exposed females
than in controls, suggesting decreased dopamine turnover (no such
effect was evident in the males: Gonzalez et al., 2003). By contrast,
the expression of the tyrosine hydroxylase (TH) gene was weaker
in the mesencephalon of exposed animals (Bonnin et al., 1994) or
unchanged in the ventrolateral medulla oblongata (Tree et al., 2014).
The aforementioned altered dopaminergic function could gener-
ate compensatory changes at the receptor level. For example, D2
receptor affinity was lower in the striatum of THC-exposed ani-
mals (Walters and Carr, 1988), and the amount of D2 receptors
in the NAcc was also reduced (DiNieri et al., 2011). More specifi-
cally, a decrease was observed in 3 H-raclopride binding in the NAcc
but not in the dorsal striatum of adult male rats exposed to THC
in utero (DiNieri et al., 2011). This alteration was produced via a
decrease in the 3meH3K4 induction mark and an increase in the
2meH3K9 repression mark in the offspring of THC-exposed moth-
ers (DiNieri et al., 2011). In functional terms, these changes in D2
receptor values seem to have a differential effect at pre- and post-
synaptic levels. Thus, perinatal exposure to THC in rats enhanced
presynaptic dopamine D2 receptor mediated responses while post-
synaptic dopamine D2 receptor agonist-induced stereotypes were
dampened. Interestingly, these results were only observed in males
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
and not in females (Moreno et al., 2003; but see Garcia et al.,
1996). Given the crucial role of D2 receptors in drug addiction,
it is tempting to speculate that the reduction in these receptors
might somehow contribute to the increased rewarding effects of
opiates observed in animals exposed to cannabinoids during peri-
natal development. Indeed, imaging studies of adult subjects with
a history of drug abuse have reported reduced levels of D2 in the
striatum (Volkow et al., 2004), and animal models documented that
increasing NAcc D2 gene expression attenuates drug intake (Thanos
et al., 2008).
Only a few studies have focused on serotoninergic alterations
following prenatal cannabinoid exposure but in general, there
seems to be a decrease in 5-hydroxytryptamine (5-HT or sero-
tonin) levels and an increase in 5-HT metabolism in animals that
were exposed to THC during perinatal development. The hypotha-
lamus seems to show the most consistent results, with a decrease
in serotonin uptake sites and increased 5-hydroxyindoleacetic acid
(5-HIAA) levels (Molina-Holgado et al., 1993). This alteration to the
5-HT system seems to occur early in development since decreased
5-HT levels in the diencephalon were evident at the end of gestation
and right after birth in animals exposed to THC, and they were more
prominent in males than in females (Molina-Holgado et al., 1996).
Moreover, maternal exposure to THC decreased the levels of 5-HT
in the adult female hypothalamus but not in the male hypothala-
mus (Dalterio et al., 1984b). In addition, cannabinol or cannabidiol
treatment during pregnancy reverted the decreased hypothalamic
levels of the neurotransmitter and metabolite in the males after
castration (Dalterio et al., 1984a). In addition to the hypothala-
mus, other brain areas also had altered 5-HT uptake sites, such as
the neostriatum (NSt) (increased), Raphe nuclei (RphN) (decreased)
and locus coeruleus (LC) (decreased: Molina-Holgado et al., 1993).
To our knowledge, no further studies have characterized the sero-
toninergic system in the adult brain after prenatal exposure to
cannabinoids. These alterations in serotoninergic system might be
related to the effects in emotional regulation reviewed above, how-
ever, the lack of consistent results at the behavioral level makes it
impossible to draw firm conclusions. If we try to compare those
studies with similar doses and administration periods, we observe
that animals show increased anxiety (Trezza et al., 2008) and
decreased 5-HT uptake sites in the RphN, which is reminiscent of
what is observed in human patients (Reimold et al., 2008).
5.2.5. Glutamatergic and GABAergic systems
Given the crucial role of glutamatergic transmission in learn-
ing, memory and synaptic plasticity, researchers have paid a
great deal of attention to changes in this neurotransmitter sys-
tem after prenatal cannabinoid treatment. There seems to be some
degree of consensus in the literature that perinatal cannabinoid
exposure dampens glutamate function in the adult animal. Using
in vivo microdialysis, a significant decrease in K+ -evoked and/or
basal extracellular glutamate levels was found in the hippocam-
pus (Mereu et al., 2003) or frontal cortex (Castaldo et al., 2007)
of juvenile and adult rats born to dams that received WIN. The
hippocampal alterations in glutamatergic transmission were trans-
lated into distorted synaptic plasticity and in this regard, the
induction of hippocampal long-term potentiation was impaired
in cannabinoid-exposed animals (Mereu et al., 2003). A similar
reduction in glutamate outflow was also observed in hippocampal
slices obtained from pups born to mothers exposed to WIN. More-
over, a reduced glutamate uptake was observed in synaptosomal
preparations, probably caused by the observed downregulation of
the glial glutamate transporters GLT1 and GLAST, and it was sug-
gested to develop as a compensatory mechanism in response to the
decreased glutamate outflow (Castaldo et al., 2010).
These decreased cortical glutamate levels could be explained
by the increased glutamate transport in this area found in rats
127
prenatally exposed to the cannabinoid agonist. Indeed, prenatal
exposure to the cannabinoid receptor agonist WIN increased glu-
tamate uptake through the overexpression of the GLT1 and EAAC1
glutamate transporter subtypes in the adult rat frontal cortex
(Castaldo et al., 2007). Cortical glutamate and norepinephrine (NE)
were also diminished in the offspring of pregnant rats exposed
to THC during pregnancy, an effect that was accompanied by
long-lasting alterations to cortical genes related to glutamatergic
and noradrenergic systems (Campolongo et al., 2007). In the
cerebellum, THC exposure during prenatal development modu-
lates glutamatergic nervous activity by diminishing glutamine
synthetase (GS) expression (Suarez et al., 2002). GS expression
increased progressively after THC withdrawal and while in males
this effect was evident right after THC cessation, in females it only
appeared after withdrawal. These reduced GS levels were taken
as an indication of reduced glutamate uptake in glial cells exposed
to THC in utero, as confirmed by the lower levels of the GLAST and
EAAC1 protein in the cerebellum of prenatally exposed rats (Suarez
et al., 2004b). There was sex-specific dynamics in this effect and
indeed, the reductions lasted long into withdrawal in the males
while they disappeared after THC cessation in the females (Suarez
et al., 2004b). Along with the dysregulated glutamatergic tone
found following in utero THC exposure, changes in glutamatergic
receptors were also observed in these rats. For example, there is a
down-regulation of the cerebellar (␣-amino-3-hydroxy-5-methyl-
4-isoxazolepropionic acid) AMPA glutamate receptor subunits
GluA1 and GluA2/3 following pre- and perinatal THC exposure.
This effect was stronger in males right after THC cessation and
even lasted 50 days after THC withdrawal (Suarez et al., 2004a).
These alterations in glutamatergic activity in cerebellar cells might
underlie the changes in intrinsic electrophysiological activity of
Purkinje neurons observed in adult rats prenatally exposed to the
cannabinoid agonist WIN (Shabani et al., 2014, 2011). Impaired
cortical N-methyl-d-aspartate (NMDA) function has also been
documented in rats exposed to THC in utero and the concentration-
dependent increase in glutamate observed in cultured cortical cells
after incubation with NMDA is lost in cells obtained from WIN-
exposed pups (Antonelli et al., 2005). As discussed, the reduced
glutamatergic tone found in these animals could be related to the
decrease in cytosolic brain derived neurotrophic factor (BDNF), as
wells as to the decrease in extracellular signal-regulated kinase
(ERK) and calcium calmodulin-kinase II phosphorylation in the
hippocampus and frontal cortex (Maj et al., 2007). Although the
evidence for cognitive impairment after perinatal cannabinoid
exposure is not wholly consistent, decreased glutamate levels in
the hypothalamus and impaired performance in a passive place
avoidance task suggest a possible link between both effects. In
addition, the decreased glutamate levels in the frontal cortex
(Castaldo et al., 2007) could be related to the poorer performance
in an attentional task and in a reversal task, both dependent on
frontal cortex function (Carli and Invernizzi, 2014; Robbins, 2007).
To our knowledge, only two studies have investigated the influ-
ence of prenatal THC exposure on GABAergic regulation during
adulthood. In the first of these studies, THC exposure did not
modify either glutamic acid decarboxylase (GAD) activity or GABA
content in the VTA, NAcc, substantia nigra, caudate-putamen or
globus pallidus, suggesting no changes in basal presynaptic activ-
ity of GABA-containing neurons (Garcia-Gil et al., 1999). The motor
response of these animals was also tested in the open-field test
after a single injection of muscimol, a GABAA receptor agonist, or
baclofen, a GABAB receptor agonist. The motor inhibition caused
by baclofen was more marked in magnitude in THC-exposed males
and females, unlike the response to the GABAA receptor agonist,
muscimol. The functional activity of GABAB receptors was also
measured to track the origins of the reported effects of baclofen
in THC-exposed animals. However, perinatal THC exposure did
128 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
not increase baclofen-stimulated [35S ]-GTP␥S binding in the areas
studied. In the second study, GAD and GABA immunoreactivity was
quantified in the cerebellum of the offspring of rats exposed to
WIN during pregnancy. Although these elements were distributed
similarly in WIN-exposed animals and their controls, there were
quantitative differences between them. WIN-treated rats had more
GAD and GABA immunoreactive cells in the molecular and Purk-
inje layers, with no changes in the granular layer (Benagiano et al.,
2007).
Clearly more research is needed to gain a deeper understanding
of the alterations to the GABAergic systems caused by cannabi-
noid exposure during perinatal development and their potential
functional implications.
5.2.6. Endogenous opioid system
One of the first pieces of evidence that perinatal cannabinoid
exposure might affect the endogenous opioid system came from an
early report showing that perinatal exposure to THC in mice altered
enkephalin and NE sensitivity in the vas deferens (Dalterio et al.,
1980). Eighteen years later, we showed that perinatal exposure
to THC produces specific long-term regional changes in ␮ opioid
receptor binding that differed in function of the sex of the animal
(Vela et al., 1998). THC-exposed males exhibited a lower density
of these receptors in the dorsal striatum and posteromedial corti-
cal amygdala than their controls. Receptor binding in THC-exposed
females was only lower than in controls in the lateral amygdala,
whereas they exhibited a higher density of these receptors in the
frontoparietal cortex, hippocampus (CA3 area), posteromedial cor-
tical amygdala, VTA and the periaqueductal gray matter. Prenatal
exposure to THC also modified the expression of genes encoding
several opioid peptides in the fetal brain, decreasing for example
PENK mRNA in the caudate-putamen, hypothalamic paraventric-
ular and ventromedial nuclei, and in the cerebral cortex of males
but increasing it in the same regions of females with the excep-
tion of the cerebral cortex (Perez-Rosado et al., 2000). Other opioid
neuropeptides have also been analyzed and prenatal-THC exposure
altered proopiomelanocortin (POMC) and PDYN mRNA expres-
sion in most brain areas studied at different fetal ages, although
the effects were sex-dependent (Perez-Rosado et al., 2002). For
example, at GD21 there was more mRNA for POMC in the arcu-
ate nucleus, cerebral cortex and habenular nuclei of THC-exposed
females and less in THC-exposed males. With regards PDYN, it was
more strongly expressed in the cerebral cortex, hippocampus and
paraventricular hypothalamic nucleus of THC-exposed females,
whereas no changes were observed in THC-exposed males. It is
important to note that these changes were evident in the fetal
brain and no studies have been carried out to determine if these
alterations are long-lasting (reaching the adult period) or transient.
With regards the functional implications of the long-term alter-
ations of ␮ opioid receptors, the most relevant finding is that their
levels decrease in the VTA of females alone, consistent with the
increased morphine self-administration observed in these animals
(Vela et al., 1998). Indeed, VTA ␮ opioid receptors play a crucial role
in opiate reinforcement (Zhang et al., 2009).
5.2.7. Endogenous cannabinoid system
The endogenous cannabinoid system is an important regulator
of several physiological and psychological functions. However, only
two studies that we are aware of have assessed the alterations to
the endogenous cannabinoid system of the adult offspring of moth-
ers exposed to THC during gestation. In the first (García-Gil et al.,
1999), no changes in cannabinoid receptor binding were witnessed
in most of the regions analyzed (basal ganglia, cerebellum, limbic
structures and most of the hippocampal regions), although some
marginal effects were found. For example, THC-exposed males but
not females exhibited a small decrease in binding in the superficial
layers of the cerebral cortex. In the CA2 layer of Ammon’s horn,
there was an increase in CB1 mRNA in THC-exposed male animals,
although the most marked and probably relevant changes were
seen in the arcuate nucleus of the hypothalamus. In this nucleus,
THC-exposed males exhibited increased binding, whereas it tended
to decrease in THC-exposed females. In the second study (Castelli
et al., 2007), CB1R density and affinity was not affected in any of
the regions analyzed, yet there was an increased functional activa-
tion of cannabinoid receptors in the hippocampus and decreased
activation in the striatum (as indicated by 35 S-GTP-␥-S binding
autoradiography in WIN-exposed offspring). Endogenous cannabi-
noid levels and cannabinoid-related enzyme activities were also
determined in this study, and the increased AEA levels in the stria-
tum were associated with reduced FAAH and enhanced NAPE-PLD
activities. Opposite changes in AEA levels and enzymatic activities
were detected in the limbic areas of WIN-treated rats (specifically
olfactory tubercles, NAcc and septal nuclei).
The increased activity of CB1Rs in the hippocampus and the
stronger transcription of its gene could suggest enhanced signal-
ing through this system, which might contribute to explain some
of the cognitive deficits reported above (Basavarajappa et al., 2014;
Mereu et al., 2003).
6. Adolescent cannabinoid exposure and long-lasting
effects on neural and psychological processes
6.1. Human studies
6.1.1. Neuropsychological assessment
As indicated, marihuana is the illegal drug most widely con-
sumed during adolescence in Europe (especially Spain, France and
Italy: European Monitoring Centre for Drugs and Drug Addiction,
2014) and the United States (Substance Abuse and Mental Health
Services Administration, 2013). Adolescence is a developmental
period of intense neural remodeling and plasticity (Blakemore,
2013; Lenroot and Giedd, 2006) with important sexual dimor-
phisms in the developmental trajectories of several brain regions
(Lenroot and Giedd, 2010; Lenroot et al., 2007; Raznahan et al.,
2014). For example, male and female adolescents show diver-
gent structural maturation within the ventro-rostral putamen,
medial rostral caudate, rostral pallidum, and lateral-dorsal tha-
lamus, which accumulate most of the functional abnormalities
associated with depression, and also the ventral striatal systems
involved in key aspects of risk-taking behavior (Raznahan et al.,
2014). Another brain region that develops substantially during
the teenage years is the PFC, which is involved in executive
functions, such as decision making, inhibitory control and plan-
ning, social understanding and self-awareness (Blakemore, 2013).
Although brain size is thought to stabilize at around five years of
age, important neurodevelopmental processes continue through-
out adolescence, including myelinization (Benes, 1989), synaptic
refinement (Huttenlocher and Dabholkar, 1997) and a reduction in
gray matter volume (Blakemore, 2012). Indeed, longitudinal stud-
ies of subjects from 3 to 30 years of age show a general pattern of
childhood peaks of gray matter followed by adolescent declines,
structural and functional rises in connectivity and integrative pro-
cessing, and a changing coordination between limbic/subcortical
and frontal lobe functions, extending well into the adult life (Giedd,
2008). As a result, insults received in adolescence might pervasively
affect the normal course of CNS development.
There has been a great deal of controversy regarding the pos-
sible long-term consequences of adolescent marihuana abuse on
cognition. Several questions arise in this debate, for example, is
the age of onset of marihuana abuse a determinant factor in its
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
detrimental effects? Second, are these deficits general or specific to
certain cognitive domains? Third, are these deficits restored after
a period of abstinence? With regard to the first question, it seems
that the age of onset is indeed an important factor. A recent study
highlighted the neuropsychological decline in persistent cannabis
intake from childhood to midlife (Meier et al., 2012). The aim of
that study was to test the association between persistent cannabis
use and neuropsychological decline, and to determine whether this
decline is concentrated among cannabis users whose consumption
begins in adolescence. The subjects belonged to the Dunedin cohort,
a birth cohort of 1037 individuals followed from birth (1972/1973)
to 38 years of age, and the strength of this work is that it included
measures of cognitive functioning prior to any known cannabis
exposure in the participants. Indeed, neuropsychological testing
was conducted at 13 years of age, before the initiation of cannabis
use, and again at 38 years. The study tried to rule out the most fre-
quently cited confounding factors in such reports (acute or residual
cannabis intoxication, tobacco dependence, hard-drug dependence
– e.g. heroin, cocaine, amphetamines, alcohol dependence, and
schizophrenia), and several deficits in neuropsychological func-
tions were observed across functional domains. Importantly, the
impairment was concentrated among adolescent-onset cannabis
users, with more persistent use associated with greater decline
(Meier et al., 2012).
Other groups have confirmed the importance of the age of onset
of cannabis use showing, for example, that early-onset cannabis
users performed worse in several neuropsychological tasks that
focused on executive functioning (Fontes et al., 2011). This was con-
firmed by relating earlier onset of marihuana use, and the increased
frequency and magnitude of marihuana use, with poorer cognitive
function relative to later onset users (Gruber et al., 2012). Another
study found that the age of onset of marihuana use was the only
factor predicting impaired visual attention (Ehrenreich et al., 1999).
Despite these findings, the issue of the importance of early onset
of cannabis abuse has been contested. It was reported that current
(but not past) heavy cannabis users performed worse than controls
in terms of verbal memory, with no differences between men and
women (Pope et al., 2001). Indeed, only current heavy cannabis
users showed a decline in IQ, memory and processing speed (but
not other abilities) at the age of 17–20 years of age relative to their
baseline performance at 9–12 years of age (Fried et al., 2002).
The second question has no clear answer. While Meier’s report
provides evidence for a general impairment, a meta-analysis of
the non-acute effects of cannabis use reported significant, yet
very modest, adverse effects of cannabis on measures of learning
and forgetting. Thus, the detrimental effects of cannabis may be
specific to some neurocognitive functions as opposed to general
(Grant et al., 2003).
The data available do not provide a definite answer to the third
question (i.e. are these deficits restored after abstinence?). For
example, it appeared that cessation of cannabis use did not fully
restore neuropsychological function (Meier et al., 2012), as con-
firmed by a recent study (Smith et al., 2015). More recently, heavy
cannabis users were shown to perform worse than controls at base-
line as well as at 1 and 7 days after supervised abstinence, but not
after 28 days (Pope et al., 2001). Clearly, more research is needed
to provide a clear answer to these questions however one of the
merits of Meier et al.’s report is to underscore the importance of
prevention and intervention efforts related to adolescent onset and
persistent regular use of cannabis (Gonzalez and Swanson, 2012).
6.1.2. Neuroimaging studies
A systematic review of the neuroimaging studies that focus
on adolescent chronic cannabis users identified two performed
on chronic adolescent cannabis users after 28 days of abstinence
(Batalla et al., 2013). In these reports, alterations in the volume of
129
the inferior posterior vermis were identified (smaller in cannabis
users), as were sex-dependent effects of cannabis use on the volume
of the PFC (larger than the controls in female cannabis users and
smaller in males). This gender-specific difference was also reported
elsewhere (McQueeny et al., 2011), with larger right amygdala
volumes in female but not male cannabis users compared to the
control population. The hippocampus is also affected by adolescent
cannabis use and in a recent study, subjects with cannabis use dis-
order that remained abstinent for six months developed alterations
to hippocampal surface shape (Smith et al., 2015).
Chronic adolescent cannabis users also seem to adopt differ-
ent strategies and engage alterative neural networks than controls
when resolving cognitive tasks. For example, cannabis users show
increased activity during a cognitive task in the basal ganglia and
postcentral gyrus of the right hemisphere and bilaterally in the
precuneus and superior parietal lobe, suggesting a compensatory
neural effort. Additionally, cannabis users demonstrated a positive
association between performance in the cognitive task and brain
activation in the left temporal gyrus, superior temporal lobe and
anterior cingulate cortex, while controls showed a negative associa-
tion in these areas. Conversely, marihuana users showed a negative
association between brain activation and cognitive performance in
the right temporal gyrus, thalamus, pulvinar, left parahippocampal
gyrus and bilateral uncus while the converse pattern was found in
controls (Padula et al., 2007). In a similar task, the performance of
adolescent cannabis users with brief and sustained cannabis absti-
nence was comparable, yet recent users displayed greater activity
in the medial and left superior prefrontal cortices and bilateral
insula. By contrast, an increased response in the right precentral
gyrus was evident in abstinent users (Schweinsburg et al., 2010).
The activation detected by fMRI in a group of abstinent cannabis
users on a Go/No-Go task has been compared with healthy subjects
(Tapert et al., 2007). Although a similar level of task performance
was achieved by both groups, adolescent cannabis users exhibited
stronger activation during inhibitory trials than healthy subjects
in the right dorsolateral prefrontal, bilateral medial frontal, bilat-
eral inferior and superior parietal lobules, and the right occipital
gyrus when. In the non-inhibitory trials, differences were mainly
found in the right prefrontal, insular and parietal cortices, with
cannabis users showing greater activation in these areas than the
controls. These results suggest a greater neurocognitive effort by
cannabis users during the task, even after the abstinence period.
Indeed, these findings might reflect a sort of neuroadaptation, i.e.
the recruitment of additional regions as a compensatory mech-
anism to maintain normal cognitive performance in response to
chronic cannabis exposure, most prominently in the PFC (Batalla
et al., 2013). In addition to all these data, the specific importance
of the age of onset of marijuana use has been recently highlighted
in a fMRI study that showed similar structural alterations in recre-
ational cannabis as those observed in heavy users (decreased gray
matter density in the left parahippocampal gyrus and the right tem-
poral lobe) only when they began consumption early in their lives
(Battistella et al., 2014).
6.2. Animal studies
6.2.1. Learning and memory
In most of the studies assessing long-term cognitive con-
sequences of adolescent cannabis exposure, the novel object
recognition task or the novel place recognition task have been used
(Abush and Akirav, 2013, 2012; Cadoni et al., 2013; Higuera-Matas
et al., 2009; Llorente-Berzal et al., 2013; Mateos et al., 2011; O’Shea
et al., 2006, 2004; O’Tuathaigh et al., 2010; Quinn et al., 2008;
Realini et al., 2011; Renard et al., 2013; Schneider and Koch, 2003;
Schneider et al., 2005; Schulz et al., 2013; Zamberletti et al., 2014,
2012), although the Y maze (Cadoni et al., 2013; Mateos et al., 2011;
130 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146

Table 5
Behavioral and cognitive alterations found in adult animals after adolescent cannabinoid exposure.

Species Sex Treatment Test age Results Reference

Drug Dose Age

Novel object recognition

Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND61, 70, 90 Memory impairment up to 10 Abush and Akirav, 2012
days after withdrawal
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND91 Memory impairment Abush and Akirav, 2013
Rat ♂ 9 -THC 2, 4 and 8 mg/kg (i.p.) PND40-42 PND70-80 No deficits shown Cadoni et al., 2013
twice a day for 1 day
each dose
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-38 PND103 No deficits shown Higuera-Matas et al., 2009
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg PND28-45 PND75 Memory impairment ♀ Llorente-Berzal et al., 2013
(i.p.), for 7, 6 and 5
days, respectively
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-43 PND76 Memory impairment ♂ Mateos et al., 2011
Rat ♀ CP 55,940 0.15, 0.2 and 0.3 mg/kg PND30-50 PND72 Memory impairment O’Shea et al., 2004
(i.p.) for 3, 8 and 10
days, respectively
Rat ♂ CP 55,940 0.15, 0.2 and 0.3 mg/kg PND30-50 PND79, 81 Memory impairment O’Shea et al., 2006
(i.p.) for 7 days each
dose
Mouse ♂–♀ 9 -THC 4 or 8 mg/kg (s.c.) PND32-52 >PND73 Memory impairment O’Tuathaigh et al., 2010
Rat ♂ 9 -THC 1 mg/kg (i.p.) for 2 days PND32-48 PND67 Memory impairment Quinn et al., 2008
and 5 mg/kg on
alternate days (8 doses)
Rat ♀ 9 -THC 2.5, 5 and 10 mg/kg PND35-45 PND98-104 Memory impairment Realini et al., 2011
(i.p.), for 3, 4 and 4
days, respectively
(twice a day)
Rat ♂ CP 55,940 0.15, 0.2 and 0.3 mg/kg PND29-49 PND77, 84 Memory impairment Renard et al., 2013
(i.p.) for 7 days at each
dose
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.). 1 dose: PND40-65 PND85-90 Memory impairment Schneider and Koch, 2003
10 days; 2 doses: 5
days; 0 doses: 10 days
(randomized)
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.). 1 dose: PND15-40 PND80 No deficits shown Schneider et al., 2005
10 days; 2 doses: 5
days; 0 doses: 10 days
(randomized)
Rat ♂ WIN 55,212-2 7.5 mg/kg (i.p.). 1 dose: PND40-65 PND75 No deficits shown Schulz et al., 2013
10 days; 2 doses: 5
days; 0 doses: 10 days
(randomized)
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg PND35-45 PND65 Memory impairment ♀ Zamberletti et al., 2012
(i.p.), for 3, 4 and 4
days, respectively
(twice a day)
Rat ♀ 9 -THC 2.5, 5 and 10 mg/kg PND35-45 >PND75 Memory impairment ♀ Zamberletti et al., 2014
(i.p.), for 3, 4 and 4
days, respectively
(twice a day)

Novel place recognition

Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND61, 70, 90, 135 Impaired spatial memory up to Abush and Akirav, 2012
75 days after withdrawal
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND91 No deficits shown Abush and Akirav, 2013
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-43 PND70 Impaired spatial memory ♀ Mateos et al., 2011
Rat ♂ CP 55,940 0.15, 0.2 and 0.3 mg/kg PND29-49 PND90, 97 Impaired spatial memory Renard et al., 2013
(i.p.) for 7 days at each
dose
Rat ♀ 9 -THC 2.5, 5 and 10 mg/kg PND35-45 >PND75 Impaired spatial memory ♀ Zamberletti et al., 2014
(i.p.), for 3, 4 and 4
days, respectively
(twice a day)

Morris water maze

Rat ♂ WIN 55,212-2 1 mg/kg (i.p.) PND30-50 >PND70 Impaired spatial learning and Abboussi et al., 2014
memory
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND61, 70 Impaired spatial memory 24 h Abush and Akirav, 2012
after withdrawal
Rat ♂ 9 -THC 5 mg/kg (i.p.) PND35-55 PND83 No deficits shown Cha et al., 2006
Rat ♂–♀ 9 -THC 5 mg/kg (i.p.) PND35-55 PND83 No deficits shown Cha et al., 2007
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-38 PND107 No deficits shown Higuera-Matas et al., 2009

Morris water maze (no-spatial)

Rat ♂ 9 -THC 5 mg/kg (i.p.) PND35-55 PND85 No deficits shown Cha et al., 2006
Rat ♂–♀ 9 -THC 5 mg/kg (i.p.) PND35-55 PND85 No deficits shown Cha et al., 2007
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146 131

Table 5 (Continued)

Species Sex Treatment Test age Results Reference

Drug Dose Age

Y maze
Rat ♂ 9 -THC 2, 4 and 8 mg/kg (i.p.) PND40-42 PND70-80 No deficits shown Cadoni et al., 2013
twice a day for 1 day
each dose
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-43 PND44 Enhanced spatial working Mateos et al., 2011
memory ♂
Mouse ♂–♀  -THC 9
4 or 8 mg/kg (s.c.) PND32-52 >PND73 Impaired spatial working O’Tuathaigh et al., 2010
memory ♀ (8 mg/kg)
Rat ♀ 9 -THC 2.5, 5 and 10 mg/kg PND35-45 Not specified Impaired spatial working Rubino et al., 2014
(i.p.), for 3, 4 and 4 memory
days, respectively
(twice a day)

Active place avoidance

Rat ♂ WIN 55,212-2 1 mg/kg (i.p.) PND30-50 >PND70 No deficits shown Abboussi et al., 2014
Rat ♂–♀ 9 -THC 2 mg/kg (i.p.) PND22-40 PND73 Impaired spatial learning and Harte and Dow-Edwards, 2010
memory
PND41-60 PND76 No deficits shown

Passive place avoidance

Rat ♂ 9 -THC 2.5, 5 and 10 mg/kg PND35-45 >PND75 No deficits shown Rubino et al., 2009
(i.p.), for 3, 4 and 4
days, respectively
(twice a day)

Radial maze
Rat ♂–♀ 9 -THC 2 mg/kg (i.p.) PND35-45 >PND75 Impaired spatial working Rubino et al., 2009
memory

Fear conditioning
Mouse ♂ WIN 55,212-2 2 mg/kg (i.p.) PND30-32, 34 and >PND120 Impaired fear conditioning Gleason et al., 2012
39

Holeboard
Mouse ♂ 9 -THC 7 mg/kg every other PND28-48 PND138 No deficits shown Tantra et al., 2014
day (i.p.)

Attentional Set-Shifting Task

Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.). 1 dose: PND40-65 PND85-100 Impaired behavioral flexibility Gomes et al., 2014
10 days; 2 doses: 5
days; 0 doses: 10 days
(randomized)

O’Tuathaigh et al., 2010; Rubino et al., 2014), the active avoidance
task (Abboussi et al., 2014; Harte and Dow-Edwards, 2010), pas-
sive avoidance (Rubino et al., 2009), the radial maze (Rubino et al.,
2009), the holeboard (Tantra et al., 2014) and the Morris water maze
(Abush and Akirav, 2012; Cha et al., 2007, 2006; Higuera-Matas
et al., 2009) have also been used. In addition, emotional mem-
ory was also examined in one study by assessing fear conditioning
memory (Gleason et al., 2012: see Table 5).
When spatial learning of adolescent rats was examined in the
water maze 28 days after chronic THC treatment, no long-lasting
cognitive alterations were observed in either male or female rats
(Cha et al., 2007, 2006). Similarly, no significant alterations in
water maze performance, neither in reference memory tests nor
working memory tests, were detected in adult male or female
rats that had been treated with the cannabinoid agonist CP during
adolescence (Higuera-Matas et al., 2009). This was further con-
firmed when water maze performance (acquisition, short-term,
long-term memory and reversal learning) was assessed in rats
exposed to the cannabinoid agonist WIN during adolescence and
after a 10 days drug-free period (Abush and Akirav, 2012). It could
be argued that the spatial component of learning and memory is
not affected by adolescent cannabinoid exposure; however, adult
rats of both sexes exposed to THC during adolescence appear to
perform worse than control animals in the radial maze (Rubino
et al., 2009). Indeed, adult rats that were exposed to WIN during
adolescence did show impaired spatial learning and memory in
the water maze (Abboussi et al., 2014). Moreover, some deficits
have also been observed in the novel place recognition task, which
has a strong spatial component (see below).
Studies using the novel object recognition task or the novel
place recognition task have shown that adolescent exposure to
cannabinoids can indeed produce cognitive sequelae in adulthood.
For example, recognition and/or place memories were impaired
in female rats (males were not tested in these experiments) that
were exposed to THC as adolescents and that were tested as adults
(Realini et al., 2011; Zamberletti et al., 2014). Similarly, two addi-
tional studies confirmed that adolescent exposure to THC was
associated with impaired performance in the novel object recogni-
tion task in adulthood, again in females (Llorente-Berzal et al., 2013;
Zamberletti et al., 2012). Notably, in Llorente-Berzal’s experiments,
the overall effects of THC in the novel object task were probably due
to a potentiated amnesic action in one of the THC groups that was
also exposed to 3,4-methylenedioxy-methamphetamine (MDMA).
Independently, a deficit in novel object recognition was also found
in adult male rats chronically exposed to THC as adolescents (Quinn
et al., 2008). These results were found in both wild type mice
and those carrying a deletion of the catechol-O-methyltransferase
(COMT) gene. Moreover, there was a general interaction between
sex and treatment showing that males were more affected than
females, yet this included a mixture of heterozygous and KO mice
making it difficult to establish a clear sexual dimorphism in the
effect of THC (O’Tuathaigh et al., 2010). Other studies also pro-
vided evidence for cognitive impairment in both male and female
adult rats (O’Shea et al., 2006, 2004). Notably, chronic exposure
132 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
to CP during adolescence induced deficits in the novel object
and place location tasks in adulthood, in a sex-dependent man-
ner (CP-exposed males showed deficits in the novel object task
while CP-exposed females performed worse in the object loca-
tion task: Mateos et al., 2011), although we failed to find evidence
for such impairments with a more restricted treatment (Higuera-
Matas et al., 2009). Deficits in both object location and novel object
tasks were also found in males when the WIN was administered,
even after several drug-free periods (1, 10, 30 and 75 days: Abush
and Akirav, 2013, 2012). These deficits seem to be specific to early
exposure, because when an equivalent WIN treatment was admin-
istered to prepubertal, adult or pubertal rats and they were tested
as adults, only the latter displayed deficits in object recognition
memory (Schneider and Koch, 2003; Schneider et al., 2005; but
see Schulz et al., 2013). Similar results were obtained in female
rats exposed to CP during adolescence but not adulthood (O’Shea
et al., 2004). It appears that the aforementioned cognitive deficits
may also depend on the genetic make-up of the individuals, in this
sense strain-specific effects have been reported in terms of the sen-
sitivity to the long-lasting amnesic actions of chronic cannabinoid
exposure during adolescence, as measured in the object location
test (Renard et al., 2013 but see Cadoni et al., 2013).
Deficits in reversal learning have also been found using the
active avoidance task. While THC administration during early ado-
lescence had no effect on acquisition, it hindered performance in
the reversal trial of both male and female rats exposed to the
cannabinoid during early but not late adolescence (Harte and Dow-
Edwards, 2010). In a recent study, deficits in a reversal task were
also found using WIN as the cannabinoid (Gomes et al., 2014). Inter-
estingly, another study using WIN reported impaired performance
in a two-way active avoidance task (Abboussi et al., 2014). How-
ever, no differences in the passive avoidance task were evident as
a result of adolescent THC exposure (Rubino et al., 2009). Emo-
tional memory is also affected by adolescent cannabinoid exposure
and contextual fear conditioning was impaired in adult mice that
had been exposed to the cannabinoid agonist WIN during adoles-
cence (Gleason et al., 2012). This effect was not evident when the
cannabinoid agonist was administered during adulthood.
From all these results summarized above, it can be concluded
that adolescent cannabinoid exposure produces deleterious effects
on recognition, emotional and spatial memory, although this is not
the case when cannabinoid exposure occurs during the prepubertal
or adult periods. In addition, while the gender specificity of these
effects is not clear, whenever a sex-difference was noted, females
showed the impairments with only one exception (Mateos et al.,
2011). In any case, further research should be carried out on this
issue.
6.2.2. Emotional regulation
There is evidence that adolescent cannabinoid exposure alters
emotional regulation in adulthood, although these results vary
depending on the particular index or process examined (see
Table 6). Thus, if we look at anxiety responses, we can find reduced
anxiety-related behavior has been observed in adulthood after
chronic adolescent cannabinoid exposure. For example, rats (both
males and females) that had been exposed to the cannabinoid ago-
nist CP during adolescence spent longer in the open arms of an EPM,
indicating reduced anxiety (Biscaia et al., 2003). Likewise, puber-
tal treatment with WIN (under an irregular administration regime)
induced an anxiolytic phenotype evident in the EPM (longer spent
in the open arms) and in the open field (higher percentage of time
spent in the central zone: Wegener and Koch, 2009). Conversely, no
alterations to measures of anxiety were evident in adulthood after
adolescence cannabinoid exposure in other studies. For example,
we treated adolescent rats with the cannabinoid agonist CP and
found no change in the time spent in the open arms of the EPM
when animals were tested as adults, suggesting no lasting changes
in anxiety (Higuera-Matas et al., 2009). This was confirmed else-
where in both the EPM and open field tests (Abush and Akirav,
2013, 2012; Bambico et al., 2010; Bortolato et al., 2014; Llorente-
Berzal et al., 2013; Mateos et al., 2011; O’Tuathaigh et al., 2010;
Rubino et al., 2008). However, a more recent report showed that
chronic THC administration during adolescence evoked an anx-
iogenic phenotype in adult animals revealed by less time spent
in the open arms of the EPM (Stopponi et al., 2014). In addition,
increased anxiety at adulthood after adolescent cannabinoid expo-
sure was reported only when anxiety was measured indirectly
in the form of anxiety-induced hyponeophagia, the inhibition of
ingestion and the approach to food when an animal is exposed
to an anxiety-provoking novel environment. Using this behavioral
paradigm, enhanced anxiety was evident in cannabinoid-exposed
rats (Bambico et al., 2010).
There seems to be a stronger agreement with regards social
anxiety, and adolescent cannabinoid treatment is reported to pro-
duce anxiogenic responses when animals reach adulthood. For
example, when social interaction was analyzed in female rats
as a measure of anxiety, CP treatment during adolescence but
not adulthood was hampered social interaction, a fact that was
interpreted as reflecting increased anxiety in CP-exposed animals
(O’Shea et al., 2004). These results in the females were confirmed
elsewhere using THC administration instead of CP (Realini et al.,
2011; but see Zamberletti et al., 2012). Interestingly, when male
rats were tested, both adolescent and adult exposure to CP gener-
ated a similar decrease in social interaction (O’Shea et al., 2006).
In part, these conflicting results may be explained by the incidence
of genetic alterations among individuals that may modulate the
organism’s response to cannabinoids. In fact, it has been shown
that a strong-weak activity COMT polymorphism may modulate
the effects of adolescent THC exposure on the risk of developing
psychosis in adulthood. Indeed, while adolescent exposure to THC
has no effects on anxiety measured in the EPM in wild-type ani-
mals, a COMT knock-out mice display decreased anxiety in this
test (O’Tuathaigh et al., 2010). Notably, this anxiolytic effect was
only evident when animals were treated as adolescents and expo-
sure to THC as adults produces no overt anxiolytic phenotype.
Another study also showed the influence of genetic background
on the effects of adolescent THC on anxiety at adulthood. By using
the Lewis and Fischer 344 rat strains as a model (see Kosten and
Ambrosio (2002) for a review), adolescent THC exposure was asso-
ciated with less anxiety in adulthood but only in the Lewis strain,
with no such effects evident in Fischer 344 rats (Cadoni et al., 2013).
Hedonic responses are another kind of emotional responses
affected by adolescent cannabinoid exposure. Anhedonia is defined
as a blunting of sensitivity to rewarding stimuli, and it is a core
symptomatological feature of depression. One way to measure
anhedonia is the sucrose preference test, where animals are given
the opportunity to choose between a sucrose solution (a natu-
ral reward in rodents) and tap water. A consistent finding is that
adolescent THC induces anhedonia at adulthood evident in this
test. For example, both male and female animals exposed to THC
as adolescents exhibited decreased sucrose preference as adults
(Rubino et al., 2008; Bambico et al., 2010; Realini et al., 2011).
This phenotype could be reverted by increasing eCB levels through
the administration of URB597, a FAAH inhibitor (Realini et al.,
2011). There is however, one report showing that sucrose pref-
erence is not affected in adult female rats exposed to CP during
adolescence (Chadwick et al., 2011), which may perhaps reflect dif-
ferences in the pharmacodynamic profiles between THC, WIN and
CP.
Although not a direct measure of depressive-like phenotype, the
FST has also been used to characterize emotional responses in adult
rats exposed to cannabinoids as adolescents. Adolescent exposure
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146 133

Table 6
Emotional behavior alterations after adolescent cannabinoid exposure.

Species Sex Treatment Test age Results Reference

Drug Dose Age

Elevated plus maze

Rat ♂ WIN 55,512-2 0.2 or 1 mg/kg (i.p.) PND70-100 >PND70 No significant Bambico et al., 2010
effects
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND35-45 PND75-84 Anxiety ↓ Biscaia et al., 2003
Rat ♂ WIN 55,212-2 1 mg/kg (i.p.) twice a day PND35-48 PND60-70 No significant Bortolato et al., 2014
effects
Rat ♂ 9 -THC 2, 4 and 8 mg/kg (i.p.) twice a PND40-42 PND70-80 Anxiety ↓ Cadoni et al., 2013
day for 1 day each dose
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-38 PND100 No significant Higuera-Matas et al., 2009
effects
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), for 7, PND28-45 PND46 No significant Llorente-Berzal et al., 2013
6 and 5 days, respectively. effects
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-43 PND64 No significant Mateos et al., 2011
effects
Mouse ♂ 9-THC 4 or 8 mg/kg (s.c.) PND32-52 PND73 No significant O’Tuathaigh et al., 2010
effects
Rat ♂–♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND75-85 No significant Rubino et al., 2008
4 and 4 days, respectively effects
(twice a day)
Rat ♂ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND52 Anxiety ↑ Stopponi et al., 2014
4 and 4 days, respectively
(twice a day)
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.). 1 dose: 10 PND40-65 PND90 Anxiety ↓ Wegener and Koch, 2009
days; 2 doses: 5 days; 0 doses:
10 days (randomized)

Open field
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND61, 70, 90 Anxiety ↓ 24 h after Abush and Akirav, 2012
withdrawal
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND91 No significant Abush and Akirav, 2013
effects
Rat ♂ WIN 55,512-2 0.2 or 1 mg/kg (i.p.) PND70-100 >PND70 No significant Bambico et al., 2010
effects
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND35-45 PND75-84 No significant Biscaia et al., 2003
effects
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), for 7, PND28-45 PND41, 75 Anxiety ↑ Llorente-Berzal et al., 2013
6 and 5 days, respectively.
Rat ♂–♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND75-85 No significant Rubino et al., 2008
4 and 4 days, respectively effects
(twice a day)
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.). 1 dose: 10 PND40-65 PND85 Anxiety ↓ Wegener and Koch, 2009
days; 2 doses: 5 days; 0 doses:
10 days (randomized)

Social interaction
Mouse ♂ WIN 55,212-2 2 mg/kg (i.p.) PND30-32, 34 and 39 PND120+ No significant Gleason et al., 2012
effects
Rat ♀ CP 55,940 0.15, 0.2 and 0.3 mg/kg (i.p.) for PND30-50 PND73 Social anxiety ↑ O’Shea et al., 2004
3, 8 and 10 days, respectively
Rat ♂ CP 55,940 0.15, 0.2 and 0.3 mg/kg (i.p.) for PND30-50 PND80 Social anxiety ↑ O’Shea et al., 2006
7 days each dose
Rat ♂  -THC
9
1 mg/kg (i.p.) for 2 days and PND32-48 PND63 Social anxiety ↑ Quinn et al., 2008
5 mg/kg on alternate days (8
doses)
Rat ♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND98-104 Social anxiety ↑ Realini et al., 2011
4 and 4 days, respectively
(twice a day)
Rat ♂–♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND68 No significant Zamberletti et al., 2012
4 and 4 days, respectively effects
(twice a day)

Forced swimming test

Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND91 No significant Abush and Akirav, 2013
effects
Rat ♂ WIN 55,512-2 0.2 or 1 mg/kg (i.p.) PND70-100 >PND70 Depressive-like Bambico et al., 2010
behavior ↑
Rat ♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND98-104 Depressive-like Realini et al., 2011
4 and 4 days, respectively behavior ↑
(twice a day)
Rat ♂–♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND75-85 Depressive-like Rubino et al., 2008
4 and 4 days, respectively behavior ♀↑
(twice a day)
Rat ♂–♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND71 Depressive-like Zamberletti et al., 2012
4 and 4 days, respectively behavior ↑
(twice a day)
134 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146

Table 6 (Continued)

Species Sex Treatment Test age Results Reference

Drug Dose Age

Sucrose preference
Rat ♂ WIN 55,512-2 0.2 or 1 mg/kg (i.p.) PND70-100 >PND70 Anhedonia ↑ Bambico et al., 2010
Rat ♀ CP 55,940 0.4 mg/kg/day (i.p.) PND35-45 PND97-99 No significant Chadwick et al., 2011
effects
Rat ♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND98-104 Anhedonia ↑ Realini et al., 2011
4 and 4 days, respectively
(twice a day)
Rat ♂–♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND75-85 Anhedonia ↑ Rubino et al., 2008
4 and 4 days, respectively
(twice a day)

Sucrose intake
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.) PND45-60 PND61, 70 and 100 No significant Abush and Akirav, 2013
effects

Palatable food preference

Rat ♀ 9-THC 2.5, 5 and 10 mg/kg (i.p.), for 3, PND35-45 PND98-104 Anhedonia ↑ Realini et al., 2011
4 and 4 days, respectively
(twice a day)

Novelty suppressed feeding

Rat ♂ WIN 55,512-2 0.2 or 1 mg/kg (i.p.) PND70-100 >PND70 Anxiety ↑ (1 mg/kg Bambico et al., 2010
dose)

Emergence test
Rat ♂ CP 55,940 0.15, 0.2 and 0.3 mg/kg (i.p.) for PND30-50 PND82 No significant O’Shea et al., 2006
7 days each dose effects

to THC increased the immobility time in adult rats (although only
in females: Rubino et al., 2008), and it was shown that this phe-
notype could be reverted by increasing eCBs levels (Realini et al.,
2011). However, these sex-specific effects were not reproduced and
increased immobility was seen in both males and females (Bambico
et al., 2010; Zamberletti et al., 2012). By contrast, no changes in
immobility time were observed in the FST on adults after late-
adolescent exposure to WIN (Abush and Akirav, 2013).
6.2.3. Sensitivity to drugs of abuse later in life and potential
addictive behavior
There is evidence that adolescent use of marihuana could
facilitate later drug abuse, a phenomenon that has been termed
the Gateway hypothesis (Kandel et al., 2006). Earlier animal
studies on psychostimulant drugs did not seem to support this
hypothesis, with adolescent exposure to THC producing no long-
lasting modulatory effects on locomotor activity or dopaminergic
responses induced by amphetamine (Ellgren et al., 2004). How-
ever, it was later shown that adolescent exposure to THC indeed
affected heroin reward in rats. In a series of elegant experiments,
increased heroin self-administration (30 mg/kg/infusion) was seen
in THC-exposed rats as compared to control animals, and there was
also an upwards shift in the dose-response curve (Ellgren et al.,
2007). This heightened opiate sensitivity in THC animals was also
associated to increased cue-induced relapse (Tomasiewicz et al.,
2012). There is however, one report suggesting that adolescent
THC is not associated to increased heroin self-administration
during adulthood (Stopponi et al., 2014). These conflicting results
could be due to the heroin dose used during training. Indeed. Stop-
poni et al., used a dose of 20 ␮g/kg, and when Ellgren et al., first
tested a dose of 15 ␮g/kg, no differences were evident between
animals receiving THC or the vehicle alone. However, signifi-
cant differences were later seen in the range of 7.5–30 ␮g/kg,
suggesting that these conflicting results may be the result of:
the different THC treatment regimes administered (1.5 mg/kg;
Postnatal day (PND) 28–49 in Ellgren et al., and Tomasiewicz et al.,
and increasing doses from 2.5 to 10 mg/kg; PND 35–45 in Stopponi
et al.); the rat strain used (Wistar vs. Long–Evans); or an interac-
tion of both factors. As mentioned above, THC exposure during
adolescence also increased cue-induced but not stress (24 h food
deprivation)-induced reinstatement (Tomasiewicz et al., 2012).
However when a pharmacological stressor (yohimbine) was used
to reinstate heroin self-administration behavior, the response
of THC exposed animals was stronger than that of the controls
(Stopponi et al., 2014). In a follow-up study, PENK was shown to
be a crucial factor mediating the enduring effects of adolescent
cannabis exposure on adult opiate vulnerability (Tomasiewicz
et al., 2012). Indeed, the enhanced heroin self-administration
induced by adolescent THC exposure was impaired when PENK
levels were decreased by viral-mediated transfer of a miRNA tar-
geting the PENK gene. The increased vulnerability to the rewarding
actions of opiates induced by adolescent cannabinoid exposure has
also been observed with CPP (Cadoni et al., 2013; Morel et al., 2009).
There seems to be some sex-dependency in these effects.
Indeed, when we exposed male and female adolescent rats to CP
during adolescence and tested their morphine self-administration
as adults, increased self-administration was only evident in males
(Biscaia et al., 2008). These effects of adolescent cannabinoid
exposure on the rewarding effects of morphine in adulthood might
be transgenerational. Female rats treated with WIN or its vehicle
for three consecutive days during adolescence (30 days of age)
were subsequently mated in adulthood (60 days of age), and the
adolescent and adult male offspring of these females were tested
for their response to morphine using the CPP paradigm (Byrnes
et al., 2012). Both adolescent and adult WIN-F1 offspring exhibited
greater sensitivity to morphine CPP than their vehicle-F1 counter-
parts. A similar effect was observed on morphine sensitization in
the adult female offspring of rats treated with WIN during adoles-
cence (Vassoler et al., 2013). These transgenerational effects were
also evident in the heroin self-administration paradigm, whereby
adolescent exposure to THC resulted in compulsive heroin intake
in the subsequent generation of rats as a consequence of parental
germline exposure to the drug (Szutorisz et al., 2014). The facili-
tating effects of adolescent THC exposure on opiate reward during
adulthood also depend on the genetic background of individuals.
Adolescent THC exposure in Lewis rats did not affect heroin CPP
but potentiated the effect of heroin priming on drug seeking. By
contrast, adolescent THC exposure to Fischer 344 rats increased
heroin CPP and made it resistant to extinction. THC-treated Lewis
rats also showed increased seeking reactions during extinction and
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
higher hedonic reactions in response to heroin priming (Cadoni
et al., 2013).
With regard to psychostimulants, although earlier reports
showed that THC exposure during adolescence produced no long-
lasting changes in the motor response to amphetamine, recent
studies provide conflicting data with an increased response to
amphetamine challenge (only in females in this one case: Gomes
et al., 2014; Lee et al., 2014). We also demonstrated a gateway
effect in female rats for cocaine self-administration in adulthood
following adolescent CP administration. Yet no differences were
observed during the maintenance phase once the behavior had
been stably acquired and this effect was not observed when a
natural reward (food pellets) was used as a reinforcer (Higuera-
Matas et al., 2008). The altered sensitivity to the initial rewarding
effects of cocaine could be explained by modifications to the brain’s
response to acute cocaine in our animals. We used positron emis-
sion tomography to monitor brain responses to an acute cocaine
challenge and found that chronic cannabinoid treatment abolished
the decrease in metabolic activity in the dorsal striatum of males
that was observed after cocaine was administered to control rats. In
addition, septal metabolism at resting conditions was higher in CP-
exposed females but it reached the basal values observed in control
female rats after cocaine injection (Higuera-Matas et al., 2011).
A recent cohort study in humans has provided evidence for a
so called “reverse-gateway effect” whereby cannabis consumption
during adolescence would be associated to increased tobacco con-
sumption later in life (Badiani et al., 2015). Animal studies are
needed to experimentally verify this association and further inves-
tigate the underlying neurobiological mechanisms.
6.2.4. Monoaminergic systems
Chronic adolescent cannabinoid exposure is associated to
dampened dopaminergic activity in adulthood and chronic ado-
lescent THC treatment reduced TH cell density in the VTA of mice
(Behan et al., 2012). Not only were there fewer dopaminergic cells
but there is also increased dopamine turnover in adult animals
exposed to cannabinoids during adolescence. Indeed, when the
dopamine, NE, and DOPAC levels as well as the DOPAC/dopamine
ratio in the mPFC, and dorsal and ventral striatum were examined,
evidence was found of disturbed dopaminergic regulation. Indeed,
a higher DOPAC/dopamine ratio (indicating an increased dopamine
turnover) was observed in both the dorsal and ventral striatum
of adult rats chronically treated with WIN (Bortolato et al., 2014).
In addition to fewer dopamine cells and increased turnover, there
also seems to be a heightened dopamine uptake. In this respect,
we found that adult female but not male rats treated with CP
during adolescence had higher dopamine transporter levels in the
dorsal striatum (Higuera-Matas et al., 2010). D1 receptors were up-
regulated in the NAcc-Shell of cannabinoid-exposed males (but not
females), while D2 receptors were down-regulated in the CA1 hip-
pocampal field in animals exposed to cannabinoids, irrespective of
sex.
In terms of the changes in receptor levels after adolescent
cannabinoid administration, more D1 receptors were found in the
NAcc of female but not male animals, and more D2 receptor levels
in the PFC of male but not female rats (Zamberletti et al., 2012).
There was also evidence of higher D2 levels in the NAcc in both
sexes (Zamberletti et al., 2012). The different results obtained in
the aforementioned studies may reflect the distinct cannabinoids
used, the different age at which treatment was administered or the
differences in the technical details of the autoradiography studies
(different ligands used or the lack of reported non-specific binding
controls). Some of the changes in the dopaminergic system might
also be transgenerational and in this respect, weaker expression
of the gene encoding the D2 receptor (Drd2A) was reported in the
135
dorsal striatum of the adult F1 offspring of rats exposed to THC in
their adolescence (Szutorisz et al., 2014).
Apart from the aforementioned changes, the responsiveness
of the dopaminergic system to pharmacological stimuli is also
affected. For example, the dynamics of the dopaminergic response
in the NAcc was not affected after THC or heroin challenges in Lewis
and Fischer 344 rats that had been exposed to THC during their
adolescence (Cadoni et al., 2013). However, the responsiveness to
heroin was altered after adolescent THC such that the increase
in dopamine after a heroin challenge was higher in THC-exposed
animals in the NAcc-core (Fischer 344 rats) or both NAcc-core
and NAcc–shell (Lewis rats). By contrast, single-unit extracellular
recordings obtained from antidromically identified mesoaccum-
bens dopaminergic neurons, and from their target cells in the NAcc,
show that dopaminergic neurons were significantly less responsive
to stimulation by WIN in animals pretreated for 3 days during ado-
lescence or adulthood with WIN and allowed a 2-week washout
(Pistis et al., 2004). However, in the group pretreated in adolescence
but not in adulthood, long-lasting cross-tolerance to morphine,
cocaine and amphetamine developed. They concluded that an
enduring form of neuronal adaptation occurs in dopaminergic neu-
rons after sub-chronic cannabinoid intake at a young age, affecting
the subsequent responses to drugs of abuse. Regarding this matter,
an increased number of spontaneously active VTA dopaminergic
neurons were seen recently in adult rats treated with WIN during
puberty in a recent study by (Gomes et al., 2014).
It is tempting to speculate that the altered function of the DA sys-
tem (especially the increased dopamine uptake and turnover, the
increased number of spontaneously active VTA dopamine neurons
and the enhanced dopamine levels after heroin) in cannabinoid
exposed animals might be responsible for the enhanced reward
properties of opiates and psychostimulants reported before. Stri-
atal D2 receptors might also play an important role as indicated by
perinatal studies, although the evidence is conflicting, with some
studies reporting increases (Zamberletti et al., 2012), others no
changes (Higuera-Matas et al., 2010) or even decreases (Szutorisz
et al., 2014).
Very little is known about the long-term effects of adolescent
cannabinoid exposure on the adult serotoninergic or adrenergic
systems. One study looked at the responses of serotoninergic and
noradrenergic neurons in adult rats exposed to WIN during ado-
lescence. Serotoninergic neural activity in the dorsal raphe was
significantly attenuated in WIN-exposed rats in a manner that was
independent of 5-HT1A receptors and that was probably caused
by dysregulation of CB1Rs (Bambico et al., 2010). When consid-
ering the mutual innervation existing between the dorsal raphe
serotoninergic neurons and LC noradrenergic neurons, the neural
activity of LC noradrenergic neurons was enhanced after chronic
adolescent (but not adult) cannabinoid treatment. This neurochem-
ical picture, i.e. dampened 5-HT function and increased NE tone, is
a good candidate to explain the increased anxiety found in some
studies (Bambico et al., 2010; Gordon and Hen, 2004; O’Shea et al.,
2006, 2004; Stopponi et al., 2014; Yamamoto et al., 2014). A more
recent set of experiments studied the levels of the serotonin trans-
porter (SERT) protein in the hippocampus and parietal cortex of
adult rats treated with THC during adolescence. In males, THC
increased the number of SERT positive fibers while no such effect
was evident in the females (Lopez-Rodriguez et al., 2014).
6.2.5. Glutamatergic and GABAergic systems
When exposed to cannabinoid as adolescents, a profound sex-
dependent disturbance of the glutamate/GABA balance in the
hippocampus of adult rats became evident (Higuera-Matas et al.,
2012). Indeed, CP-treated females exhibit increased GABAA recep-
tor levels, as well as decreased K+ -induced glutamate release and
NMDA receptor levels. These females also showed weaker GABA
136 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
transporter 1 gene expression (GAT-1), and increased K+ -induced
GABA release, however this effect was also evident in CP-treated
males. These results suggest enhanced GABAergic and decreased
glutamatergic tone in the hippocampus of these female rats. Very
similar results were reported for NMDA receptors, whereby female
but not male rats exposed to THC during adolescence had fewer
NMDA receptors in the hippocampus as adults (Zamberletti et al.,
2012). Similarly, fewer NMDA receptors were detected in the hip-
pocampus of adult male rats (females were not studied in this
work) exposed to cannabinoids during adolescence (Rubino et al.,
2009), an effect that was replicated in the dorsal striatum in
the F1 offspring of the rats exposed to THC during adolescence
(Szutorisz et al., 2014). Different subunits of the NMDA and AMPA
glutamate receptors were also studied in these rats and weaker
expression of the Grin1, Grin2A, Gria1 and Gria2 genes (encoding
the GluN1, GluN2B, GluA1 and GluA2 glutamate receptor subunits,
respectively) was found in the striatum of the offspring of the THC-
exposed rats. When the protein expression of these receptors was
assayed in western blots, only GluN1 and GluN2B subunits were
diminished in THC exposed animals (Szutorisz et al., 2014). The
effects of adolescent THC exposure on the glutamatergic system
have also been studied in the PFC of female rats. In this case, GluN2B
and GluA1 levels at adulthood were higher in THC-exposed rats
(Rubino et al., 2014). In addition, mGlu5 protein levels were dimin-
ished in the hippocampus of adult mice exposed to WIN during
adolescence (Gleason et al., 2012), although we could not replicate
these findings when CP was the cannabinoid agonist administered
to adolescent rats (Higuera-Matas et al., 2012). PSD95, an impor-
tant scaffolding protein of the post-synaptic density (reviewed in
Van Zundert et al., 2004), was diminished in the hippocampus of
THC-exposed male rats (Rubino et al., 2009), yet it increased in the
PFC of female rats after THC treatment during adolescence (Rubino
et al., 2014) which is consistent with the increased levels of the
glutamatergic subunits mentioned above.
The decrease in glutamate function in the hippocampus (i.e.
decreased glutamate levels after depolarization, decreased NMDA
receptor levels, decreased PSD95 levels, etc.) is consistent with
the learning and memory impairments found in these animals,
although to our surprise, there were not consequences on the
establishment of long-term potentiation (LTP) in the hippocampus
(Higuera-Matas et al., 2009), perhaps due to the saturating pro-
tocol used for LTP induction. Indeed, other authors were able to
observe LTP deficits in the ventral subicular-NAcc pathway (Abush
and Akirav, 2013, 2012).
With regard GABAergic cells, increased parvalbumin-positive
cell soma sizes have been detected in the PFC of adult animals
exposed to THC as adolescents with no changes in the density of
these cells, suggesting a disturbance of prefrontal GABA function
in THC-exposed animals (Behan et al., 2012). By contrast, fewer
parvalbumin- and cholecystokinin-positive GABAergic cells were
found in the PFC (Zamberletti et al., 2014), although the total par-
valbumin or cholecystokinin protein assayed in western blots was
no different in THC and control animals. Differences in the kind of
animals used (mice or rats), the sex (males in the former exper-
iments and females in the latter report) or some combination of
both might account for the differences in the results between these
experiments. The levels of GABA were also measured and in the
PFC, lower basal GABA levels were obtained in female rats (males
not tested) that had been exposed to THC during adolescence, a
fact that can be explained by the lower levels of GAD67 (the GABA
synthetic enzyme), in this structure (Zamberletti et al., 2014). The
functional correlate of this altered prefrontal GABA function is still
to be determined but some candidate events have been proposed.
For example, in a series of elegant experiments a history of WIN
exposure during early or mid-adolescence, but not in late adoles-
cence or adulthood, was sufficient to yield a state of prefrontal
disinhibition in adulthood comparable to that seen in the juvenile
PFC (Cass et al., 2014). Such disinhibition could be rescued by local
infusion of a GABAA receptor agonist, suggesting that GABAA recep-
tors may be downregulated in PFC neurons as a consequence of
pubertal WIN exposure (Cass et al., 2014).
6.2.6. Endogenous opioid system
Adolescent THC exposure leads to long-term discrete mesolim-
bic and nigrostriatal alterations in elements of the endogenous
opioid system. Adult male rats treated with THC during adoles-
cence displayed higher levels of PENK mRNA in the NAcc-Shell
(Ellgren et al., 2007; Tomasiewicz et al., 2012). In addition, these
authors found evidence for increased ␮ opioid receptor activity in
the substantia nigra and VTA, although binding to ␮ opioid recep-
tors was unaffected in this study (Ellgren et al., 2007). All these
changes in the endogenous opioid systems could be correlated with
the enhanced heroin self-administration and relapse observed in
THC-exposed rats (Ellgren et al., 2007; Tomasiewicz et al., 2012).
By contrast, no changes in ␮ opioid receptor levels or activity were
detected in a study that also reported decreased PENK gene expres-
sion in the dorsal striatum and NAcc-Shell (Morel et al., 2009).
However, when the expression of the ␮ opioid receptor was tested
after a morphine challenge in a sensitization paradigm, the female
offspring of rats treated with WIN as adolescents expressed more ␮
opioid receptor (OPRM1) mRNA in the NAcc (Vassoler et al., 2013).
An effect of adolescent THC on adult levels of the dynorphin peptide
was also found in the NAcc of female (but not male) rats (Rubino
et al., 2008). Increased dynorphin levels were found in THC-exposed
rats, which were related to the emotional disturbances provoked by
the cannabinoid treatment. We also analyzed the levels and activ-
ity of ␮ opioid receptors in several brain areas of male and female
rats exposed to CP during adolescence. We found that CP-treated
rats (both males and females) displayed more ␮ opioid receptors
in the sub-callosal streak. We also observed significant interactions
between the sex of the animals and cannabinoid exposure in several
brain regions, such as the cingulate cortex, CA2 and CA3 hippocam-
pal fields and several thalamic nuclei, whereby males exposed to
CP showed an overall decrease while females exposed to CP dis-
played an overall increase in the same regions. In addition, the
activity of these receptors was lower in the NAcc-Shell of CP-male
but not female rats which could be related to the increased mor-
phine self-administration evident in CP-exposed animals (Biscaia
et al., 2008).
6.2.7. Endogenous cannabinoid system
The long-lasting effects of cannabinoid administration during
adolescence on the endogenous cannabinoid system later in life
remain controversial. While there are studies where no long-lasting
changes in immunoreactivity, binding to or functional activation of
cannabinoid are reported (Behan et al., 2012; Chadwick et al., 2011;
Ellgren et al., 2007; Gleason et al., 2012; Higuera-Matas et al., 2012;
Lopez-Rodriguez et al., 2014; Morel et al., 2009), other reports
describe lower levels of these receptors in adult animals admin-
istered a cannabinoid during adolescence in almost all the areas
studied (Rubino et al., 2014, 2008; Zamberletti et al., 2012). For
these reason it is not possible to draw firm conclusions on the func-
tional relevance of these changes. Thus, in one study using CP as
the cannabinoid agent, a down-regulation in the levels of CB1Rs
in some hippocampal areas was observed in males while an up-
regulation was detected in the females (López-Gallardo et al., 2012).
Likewise, a sustained increase in cannabinoid levels during ado-
lescence (generated by blocking the FAAH enzyme with URB597)
was also associated to lower CB1R levels in the hippocampus, cau-
date putamen, NAcc, VTA and higher levels in the locus coeruleus
(Marco et al., 2009). However, in a later study where THC was the
cannabinoid agonist administered during adolescence, no changes
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146 137

Table 7
Neurochemical alterations found in adult animals after adolescent cannabinoid exposure.

Species Sex Treatment Tests Reference

Drug Dose Age Age Results

Monoaminergic systems
Rat ♂ WIN 55,512-2 0.2 or 1 mg/kg (i.p.) PND30-50 >PND70 5-HT neurons activity: DRph ↓; NA Bambico et al., 2010
neurons activity: LC (1 mg/kg dose)
↑
Mouse ♂ 9 -THC 8 mg/kg (s.c.) PND32-52 PND150- TH+ cells density: VTA ↓ Behan et al., 2012
160
Rat ♂ WIN 55,212-2 2 mg/kg (i.p.) PND35-48 PND60-70 DOPAC levels: CPu ↑; DOPAC/DA Bortolato et al., 2014
ratio: NAcc ↑, CPu ↑
Rat ♂ 9 -THC 2, 4 and 8 mg/kg (i.p.) twice PND40-42 PND63-77 DA response to heroin: NAccCo ↑; Cadoni et al., 2013
a day for 1 day each dose NAccSh ↑a
Rat ♂ WIN 55,212-2 1.2 mg/kg (i.p.). 1 dose: 10 PND40-65 PND85-100 Locomotor response to Gomes et al., 2014
days; 2 doses: 5 days; 0 amphetamine ↑
doses: 10 days
(randomized)
>PND100 Spontaneously active DA neurons:
VTA ↑
Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-38 PND121 DAT levels: CPu ↑♀; D1 receptor Higuera-Matas et al., 2010
density: NAccSh ↑♂; D2 receptor
density: HP(CA1-rad) ↓
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg, for 7, 6 PND28-45 PND89-92 SERT+ fibers: PaCx ↑♂ Lopez-Rodriguez et al., 2014
and 5 days, respectively
(i.p.)
Rat ♂ WIN 55,512-2 2, 4 and 8 mg/kg (i.p.) (one PND38-40 PND54 WIN and Morphine evoked firing Pistis et al., 2004
day each) rate and burst firing: MsLimbPth ↓;
Cocaine and Amphetamine evoked
firing rate: MsLimbPth ↓
Rat ♂ 9 -THC 1.5 mg/kg (i.p.) PND28-49 PND62 (F1 D2 receptor mRNA: CPu ↓(F1) Szutorisz et al., 2014
offspring)
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND82 D1 receptor density: NAcc ↑♀; D2 Zamberletti et al., 2012
for 3, 4 and 4 days, receptor density: PFC ↑♂; NAcc ↑
respectively (twice a day)

Glutamatergic and GABAergic systems

Mouse ♂ WIN 55,512-2 8 mg/kg (s.c.) PND32-52 PND150- Parvalbumine cell soma size: PFC ↑ Behan et al., 2012
160
Rat ♂ WIN 55,512-2 2 mg/kg (i.p.) PND35-40 PND65-85 IPSC frequency: mPFC (layer V) ↓ Cass et al., 2014
Mouse ♂ WIN 55,512-2 2 mg/kg (i.p.) PND30- PND150 mGlu5 levels: HP ↓ Gleason et al., 2012
32/33/34/39
Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-38 PND100- K+ evoked GABA levels: HP ↑; K+ Higuera-Matas et al., 2012
115 evoked Glu levels: HP ♀↓; GAT-1
mRNA: HP ↓;
GABAA receptor density: HP ♀↑;
GABAB receptor density: HP ↑;
NMDA receptor density: HP ♀↓
Rat ♂ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND75 NMDA receptor density: HP ↓; Rubino et al., 2009
for 3, 4 and 4 days, PSD95 levels: HP ↓
respectively (twice a day)
Rat ♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND46, 60 GluN2A levels: PFC ↑ (PND60); Rubino et al., 2014
for 3, 4 and 4 days, and 75 GluN2B and GluA1 levels: PFC ↑
respectively (twice a day) (PND75); PSD95 levels: PFC ↑
Rat ♂ 9 -THC 1.5 mg/kg (i.p.) PND28-49 PND35 (F1 Grin2A and Gria2 mRNA: NAcc ↑ Szutorisz et al., 2014
offspring)
PND62 (F1 Grin1, Grin2A, Gria1 and Gria2
offspring) mRNA: CPu ↓; GluN1 and GluN2B
levels: CPu ↓; NMDA receptor
density: CPu ↓
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND82 NMDA receptor density: HP ↓♀ Zamberletti et al., 2012
for 3, 4 and 4 days,
respectively (twice a day)
Rat ♀ 9 -THC 2, 5, 5 and 10 mg/kg (i.p.) PND35-45 PND75 GAD67 levels and activity; PFC ↓; Zamberletti et al., 2014
for 3, 4 and 4 days, GABA levels: PFC ↓;
respectively (twice a day) GAD67+ /Parvalbumin+ cells: PFC ↓;
GAD67+/Cholecystokinin+ cells:
PFC ↓

Endogenous opioid system

Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND35-45 PND84 ␮ opioid receptor density: SCst ↑; Biscaia et al., 2008
CinCx ♂↓♀↑; HC ♂↓♀↑; Th ♂↓♀↑;
␮ opioid receptor function: NAccSh
↓♂
Rat ♂ 9 -THC 1.5 mg/kg (i.p.) every third PND28-49 PND57 ␮ opioid receptor density: No Ellgren et al., 2007
day significant alterations found; ␮
opioid receptor function: SN ↑;
VTA ↑; PENK mRNA: NAccSh ↑
138 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146

Table 7 (Continued)

Species Sex Treatment Tests Reference

Drug Dose Age Age Results

Rat ♂ Dronabinol (9 -THC) 5 or 10 mg/kg (i.p.). Days 1, PND35-48 PND104 PENK mRNA: NAccSh ↓; CPu ↓ Morel et al., 2009
3, 4 and 6: 1 dose; days 7, 9
10, 11, 13 and 14: 2 doses;
days 2, 5, 8 and 12: No dose
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND46 Dynorphin A levels: NAcc ♀↑ Rubino et al., 2008
for 3, 4 and 4 days,
respectively (twice a day)
Rat ♂ 9 -THC 1.5 mg/kg (i.p.) every third PND28-49 PND79 PENK mRNA: NAccSh ↑ Tomasiewicz et al., 2012
day

Endogenous cannabinoid system

Mouse ♂ 9 -THC 8 mg/kg (s.c.) PND32-52 PND150- CB1 receptor density: No Behan et al., 2012
160 significant alterations found in any
of the areas studied
Rat ♀ CP 55,940 0.4 mg/kg (i.p.) PND35-45 PND46 and CB1 receptor density: No Chadwick et al., 2011
74 significant alterations found in any
of the areas studied
Rat ♂ 9 -THC 1.5 mg/kg (i.p.) every third PND28-49 PND57 CB1 receptor density and function: Ellgren et al., 2007
day No significant alterations found in
any of the areas studied
Mouse ♂ WIN 55,512-2 2 mg/kg PND30- PND150 MGL levels: HP ↑; FAAH levels: HP Gleason et al., 2012
32/33/34/39 ↑
Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-38 PND100- CB1 receptor function: HP (CA1, Higuera-Matas et al., 2012
115 CA2, CA3 and DG) ↑
Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-42 PND80 CB1 receptor density: HP(CA1-or) López-Gallardo et al., 2012
♂↓♀↑; HP(CA1-rad) ♂↓;
HP(CA1-lac) ♂↓; DG-pol ♂↓♀↑
Rat ♂–♀ Dronabinol (9 -THC) 2.5, 5 and 10 mg/kg, for 7, 6 PND28-45 PND89-92 CB1 receptor density: No Lopez-Rodriguez et al., 2014
and 5 days, respectively significant alterations found in any
(i.p.) of the areas studied
Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-43 PND81 CB1 receptor function: HP (CA1 and Mateos et al., 2011
CA2) ♂↑
Rat ♂ 9
Dronabinol ( -THC) 5 or 10 mg/kg (i.p.). Days 1, PND35-48 PND104 CB1 receptor density and function: Morel et al., 2009
3, 4 and 6: 1 dose; days 7, 9 No significant alterations found in
10, 11, 13 and 14: 2 doses; any of the areas studied
days 2, 5, 8 and 12: No dose
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND46 CB1 receptor density: PFC ↓; CPu Rubino et al., 2008
for 3, 4 and 4 days, ♀↓; NAcc ♂↓; HT ↓; HP ↓; Amy ↓;
respectively (twice a day) Th ↓; PAG ↓; SN ♀↓; VTA ↓; Cb ↓
CB1 receptor function: PFC ♀↓; CPu
♀↓; NAcc ♀↓; GP ♀↓; HP ↓; Amy↓;
SN ↓; VTA ♀↓; Cb ↓
Rat ♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND46, 60 CB1 receptor density: PFC ↓; CB1 Rubino et al., 2014
for 3, 4 and 4 days, and 75 receptor function: PFC ↓(PND46)
respectively (twice a day) ↑(PND60); AEA levels: PFC ↓;
MAGL activity: PFC ↓(PND46)
Rat ♂ 9 -THC 1.5 mg/kg (i.p.) PND28-49 PND35 (F1 CB1 receptor mRNA: NAcc ↑ Szutorisz et al., 2014
offspring)
PND62 (F1 CB1 receptor mRNA: CPu ↓
offspring)
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND82 CB1 receptor density: PFC ↓; CPu ↓; Zamberletti et al., 2012
for 3, 4 and 4 days, NAcc ↓; HT ↓; HP ↓; Th ↓; Amy ↓;
respectively (twice a day) GP ↓; SN ↓; VTA ↓; Cb ↓
CB1 receptor function: PFC ↓; CPu
↓; NAcc ↓; HT ↓; HP ↓; Th ↓; Amy
↓; GP ↓; SN ↓; VTA ↓; Cb ↓

Cholinergic system
Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-43 PND81 nACh receptor density: No Mateos et al., 2011
significant alterations found

Other signaling molecules or brain metabolism

Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-38 PND101 Basal metabolic activity: PFC ♀↑; Higuera-Matas et al., 2008
Amy-EntCx ♀↓
Rat ♂–♀ CP 55,940 0.4 mg/kg/day (i.p.) PND28-38 PND100 PSA-NCAM: HP ♂↑ Higuera-Matas et al., 2009
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND28-45 PND89-92 Arc levels: PFC ♀↓; HP ↓ Llorente-Berzal et al., 2013
for 3, 4 and 4 days,
respectively (twice a day)
Rat ♂–♀ CP 55,940 0.4 mg/kg (i.p.) PND28-42 PND80 BDNF levels: HP(CA3): ♀↓ López-Gallardo et al., 2012
Rat ♂–♀ 9 -THC 2.5, 5 and 10 mg/kg (i.p.), PND35-45 PND46 CREB activity: PFC ♀↓, HP ♀↓; NAcc Rubino et al., 2008
for 3, 4 and 4 days, ♀↑
respectively (twice a day)
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
were detected and only when MDMA was co-administered during
adolescence did THC reduce CB1R expression in the hippocam-
pus, an effect that was restricted to female rats (Lopez-Rodriguez
et al., 2014). The activity of cannabinoid receptors was also found to
be down-regulated in two of the aforementioned studies (Rubino
et al., 2008; Zamberletti et al., 2012), and up-regulated in three
other studies (in Mateos et al., who only found changes in the
hippocampus of males (Mateos et al., 2011), in Rubino et al., who
reported the alterations in the PFC of young adult female rats (males
were not tested) (Rubino et al., 2014) and in Higuera-Matas et al.,
who found increased functional activation in females but not males
(Higuera-Matas et al., 2012)). Interestingly, increments in receptor
density or function were reported in most of the studies using a syn-
thetic cannabinoid instead of THC, while decreases were reported
when the cannabinoid agent used during adolescence was THC (see
Table 7). Adolescent WIN treatment also provoked an increment in
hippocampal levels of the degrading enzymes of AEA (FAAH) and
2-AG (MAGL) (Gleason et al., 2012) although their activity was not
altered in the PFC of adult female rats exposed to THC as adolescents
(Rubino et al., 2014). Surprisingly even though the activity of FAAH
was not altered, adolescent THC exposure induced a reduction in
the levels of AEA in the PFC of female rats measured at different
time points (PNDs 46, 60 and 75: Rubino et al., 2014). Recently,
adolescent exposure to THC was shown to have transgenerational
effects on the expression of the CB1R gene. In these experiments,
CB1R gene expression was stronger in the NAcc of the adolescent
offspring of rats treated with THC as adolescents. By contrast, the
adult offspring of rats exposed to THC as adolescents had weaker
CB1R gene expression in the dorsal striatum (Szutorisz et al., 2014).
6.2.8. Cholinergic system
To our knowledge there is only one study that examined the
adult cholinergic system in rats of both sexes after chronic cannabi-
noid exposure during adolescence (Mateos et al., 2011), failing to
identify changes in nicotinic receptors in the hippocampus of these
animals.
6.2.9. Other signaling molecules and brain metabolism
Two molecules that are very important for plasticity have been
examined, phosphorylated cAMP response element-binding pro-
tein (CREB) and BDNF. In one such study, the levels of phosphoCREB
were measured in the PFC, NAcc and hippocampus of adult male
and female animals administered THC as adolescents (Rubino et al.,
2008), demonstrating a decrease in CREB phosphorylation in the
PFC and hippocampus, coupled with increased CREB activation in
the NAcc. These changes only occurred in the females and they were
related to the depressive phenotype found in female rats chroni-
cally exposed to THC as adolescents. In addition, the levels of BDNF
in CA3 were seen to decrease in adult female but not male rats
exposed to CP during adolescence (López-Gallardo et al., 2012),
which may also be relevant for the depressive phenotype described
in these animals (Rubino et al., 2008). Another plasticity-related
molecule that was down-regulated in the frontal cortex (females)
or hippocampus (males and females) after adolescent exposure to
THC was activity-regulated cytoskeleton-associated protein (Arc)
(Llorente-Berzal et al., 2013), a protein that is rapidly modulated in
a synaptic activity-dependent manner (Korb and Finkbeiner, 2011).
Finally, a recent report suggested that chronic adolescent THC expo-
sure reduced the levels of the cell-adhesion molecule NCAM 180
in the hippocampus of St8sia2 (a polysialyltransferase that trans-
fers polysialic acid molecules to NCAMs) KO mice (Tantra et al.,
2014). Given that cell adhesion molecules play a crucial role in
learning and memory (Lopez-Fernandez et al., 2007; Ronn et al.,
2000; Venero et al., 2006), these effects might explain some of the
long-term cognitive impairments found in adult animals with ado-
lescent cannabinoid exposure and a susceptible phenotype. The
139
effects of adolescent cannabinoid treatments on NCAMs seem to
be dependent on the cannabinoid agent used. In fact, when we
analyzed the levels of the different isoforms of NCAM in the hip-
pocampus after chronic adolescent CP treatment, no differences
were observed (Higuera-Matas et al., 2009). However, the levels
of the polysialylated form of NCAM (PSA-NCAM) increased in CP-
treated male rats, which are consistent with the increased levels of
polysialic acid observed in the DG of mice exposed to THC as adoles-
cents (Tantra et al., 2014). PSA-NCAM, through its anti-adhesive and
insulating properties, may participate in the remodeling of synaptic
structure, and for this reason it is crucial for brain plasticity (Nacher
et al., 2013).
In an attempt to reveal the long-term alterations in brain func-
tion caused by adolescent cannabinoid exposure, we conducted a
positron emission tomography study on adult males and females
treated with CP as adolescents. We found 18 [F]-fluorodeoxyglucose
imaging revealed a higher prefrontal cortical activity and reduced
activity in the amygdalo-enthorinal cortex of CP-exposed females.
No changes were detected in the males (Higuera-Matas et al., 2008).
6.2.10. Structural and synaptic plasticity
Long-term reduction in dendritic length, total dendritic number
and dendritic spine number have been described in the DG of adult
male rats treated with THC during adolescence (Rubino et al., 2009).
Consistent with these findings, and with the changes described
in the glutamatergic systems (see above), adolescent cannabinoid
exposure has been seen to produce long lasting impairments in
long-term potentiation of the ventral subicular-NAcc pathway (Hila
Abush and Akirav, 2012, 2013; but see Higuera-Matas et al., 2009).
Hippocampal neurogenesis is also impaired in male but not female
rats after chronic exposure to HU-210 during adolescence (Lee et al.,
2014), as confirmed using WIN (although only studied male rats
were studied) (Abboussi et al., 2014). This deficit in neurogenesis
could be due to the impaired LTP mentioned before. Indeed, it has
been shown that an LTP-inducing protocol activated a latent pre-
cursor pool, leading to increased neurogenesis in the dentate gyrus
(Kameda et al., 2012). In adult female rats, THC exposure during
adolescence abolished eCB-mediated long-term depression in the
PFC and a reduction in the spine density on distal basal dendrites
of pyramidal neurons in layer II/III in the PFC of these animals was
observed (Rubino et al., 2014).
Adolescent cannabinoid exposure not only affects basic aspects
of neural plasticity, such as LTP or spine density but also, it pervades
more general aspects of nervous function. Indeed, there is a perma-
nent suppression of cortical oscillations in mice after adolescent
exposure to cannabinoids (Raver and Keller, 2014).
These deficiencies in both structural and synaptic plasticity as
well as in cortical function might explain the aforementioned cog-
nitive sequelae induced by adolescent cannabinoid consumption.
6.2.11. Cannabinoid exposure during adolescence as a
normalizing agent against the deleterious effects of life challenges
We would like to bring up one aspect of adolescent cannabi-
noid exposure that has not been covered by earlier reviews on this
topic. During the last six years, some evidence has accumulated
in favor of possible compensating effects of adolescent cannabi-
noid exposure on the negative consequences of previous negative
events (such as maternal separation, chronic stress or chronic expo-
sure to ‘ecstasy’ (MDMA)). For instance, cannabinoids ameliorate
the impaired synaptic plasticity and short-term memory induced
by chronic stress (Abush and Akirav, 2013). The deleterious effects
of chronic immobilization stress during adolescence on spatial
memory were reverted by administration of WIN after each stress
episode, an effect that was paralleled by restoration of LTP in the
subiculum-NAcc pathway.
140 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
During adolescence cannabinoids could also be beneficial for
subjects with predisposition to show psychotic disorders. In this
regard, adolescent THC was able to compensate the increase in
aggressive behavior observed after maternal deprivation in female
but not male rats. The adolescent exposure to cannabinoids was
also able to normalize the disturbed levels of NMDA glutamater-
gic and D2 dopaminergic receptors in the dorsal striatum (only in
females), and those of D1 dopaminergic receptors in the PFC (only
in males) in adult animals subjected to a single episode of mater-
nal deprivation (Zamberletti et al., 2012). In addition, CP treatment
during adolescence also normalized the levels of CB1Rs receptors
in the hippocampus of adult rats that suffered an episode of mater-
nal deprivation (López-Gallardo et al., 2012). Similarly, chronic
treatment with WIN during adolescence attenuated the locomo-
tor response to amphetamine in rats treated in utero (GD17) with a
single dose of the DNA methylating agent methylazoxymethanol
acetate (Gomes et al., 2014). This treatment, as well as mater-
nal deprivation, is currently regarded as valid animal models of
schizophrenia (Moore et al., 2006). Cannabinoid exposure dur-
ing adolescence also abolished the increased sensitivity to the
rewarding effects of morphine induced by chronic stress during
adolescence, an effect that was accompanied by a normalization of
the altered PENK gene expression in the striatum that results from
maternal deprivation (Morel et al., 2009). Finally, chronic treatment
with WIN during puberty was able to revert the long-term impair-
ments in working memory (assessed in the novel object recognition
task) induced by chronic treatment with 3,4-methylenedioxy-N-
methylamphetamine (MDMA) during puberty (Schulz et al., 2013).
These protective actions could be related to the fact that THC can
normalize the decreased SERT levels induced by adolescent MDMA
exposure (Lopez-Rodriguez et al., 2014).
There are several questions that need to be addressed regard-
ing these protective effects. For example, is there a specific time
window for the administration of cannabinoids? What are the
downstream molecular mechanisms through which cannabinoids
exert their protective effects? What kind of insults might be
compensated by cannabinoids? There are no straight answers
to these questions. For example, WIN administered right after
stress episodes during adolescence reverted memory deficits and
restored LTP tested 30 days after the last stress episode (Abush and
Akirav, 2013). However, these protective effects are not restricted
to the adolescent period. Robust evidence for these protective
effects was also provided when the stress episode and cannabinoid
administration occur in the adult life (Ganon-Elazar and Akirav,
2009; Korem and Akirav, 2014; Segev et al., 2014). With regard to
the neurobiological mechanisms involved, the protective effects of
cannabinoids after stress during adolescence were prevented by
administering the CB1R antagonist AM251, indicative of the cru-
cial involvement of the CB1R (Abush and Akirav, 2013). Another
plausible cannabinoid effector is the glucocorticoid receptor (GR)
(Ganon-Elazar and Akirav, 2009). However, the administration of
chronic WIN together with stress exposure did not affect GRs in any
of the brain areas examined, suggesting that the beneficial effects
of WIN on memory and plasticity were not mediated by alterations
to GR in this paradigm (Abush and Akirav, 2013).
Some maternal deprivation effects are also reverted by chronic
cannabinoid administration during adolescence, such as aggres-
sive behavior or NMDA, D2 or D1 receptor levels (López-Gallardo
et al., 2012; Zamberletti et al., 2012). Why these changes only
occur in maternally deprived rats is currently unknown, but
it could be related to the long-lasting alterations of the eCB
system observed in these animals and/or (in contrast to what
is observed for chronic adolescent stress) the persistent alter-
ations in the hypothalamic–pituitary–adrenal axis (see Marco
et al., 2015 for an excellent review of the maternal deprivation
model).”
From these studies, it can be concluded that while cannabinoid
exposure during adolescence or puberty might have pervasive and
negative effects on cognition or sensitivity to the rewarding effects
of drugs of abuse, it is also true that it might have protective effects
against the deleterious consequences of other (and some times
common) challenges during adolescence or early life (such as stress,
exposure to other drugs of abuse or maternal separation).
7. Concluding remarks and future directions
In this review we have summarized the current state-of-the-art
regarding the long-lasting effects of cannabinoid exposure on cog-
nition, emotion, neurochemistry and plasticity during two critical
periods: prenatal development and adolescence. The most con-
sistent findings suggest that early cannabinoid exposure leads to
cognitive dysfunction and alterations in glutamatergic, dopamin-
ergic, cannabinoid and opioidergic transmission, with notable
sexual dimorphisms. In addition, a facilitating effect on later drug
self-administration has been observed. While the origin of such
dimorphism is not clear, it could be related to the sex differ-
ences in the eCB system seen during development (Rodriguez
de Fonseca et al., 1993), especially in the studies of perinatal
exposure to cannabinoids. Notwithstanding, cannabinoid exposure
during adolescence may also have a protective role when individ-
uals have faced severe life challenges, such as intense stress or
maternal separation. Clearly more research is needed, employing
more sophisticated behavioral measures to further define com-
plex cognitive operations, assays of self-control and impulsivity,
and paradigms that better resemble drug addiction phenomena in
humans (including the cardinal features of this disease, such as
compulsivity). Such studies would have greater external validity,
thereby better supporting preventive approaches for marihuana
consumption during pregnancy or adolescence. In addition, the
protective role of cannabinoids during puberty should be further
explored so that we can better identify individuals who might ben-
efit from the positive effects of such treatments, or those who might
suffer from their deleterious consequences.
Acknowledgements
We would like to thank all the past and present members of
the Psychobiology of Drug Addiction laboratory for their work and
insight, all of which has made this review possible. Part of the
research of our group described here was funded by: the Minis-
terio de Ciencia e Innovación (SAF2013-47520-P); the Ministerio
de Sanidad, Servicios Sociales e Igualdad (Red de Trastornos Adic-
tivos, RTA-RD12/028/0020 of Instituto de Salud Carlos III, and
Plan Nacional sobre Drogas, 2012I057); the Dirección General de
Investigación de la Comunidad de Madrid (S-2011/BMD-2308;
Programa de Actividades I+D+I CANNAB-CM); the UNED (Plan
de Promoción de la Investigación); and the European Union
(JUST/2013/DPIP/AG/4823-EU MADNESS).
References
Abboussi, O., Tazi, A., Paizanis, E., El Ganouni, S., 2014. Chronic exposure to
WIN55,212-2 affects more potently spatial learning and memory in adolescents
than in adult rats via a negative action on dorsal hippocampal neurogenesis.
Pharmacol. Biochem. Behav. 120, 95–102, http://dx.doi.org/10.1016/j.pbb.2014.
02.014
Abush, H., Akirav, I., 2013. Cannabinoids ameliorate impairments induced by chronic
stress to synaptic plasticity and short-term memory. Neuropsychopharmacol-
ogy 38, 1521–1534, http://dx.doi.org/10.1038/npp.2013.51
Abush, H., Akirav, I., 2012. Short- and long-term cognitive effects of chronic cannabi-
noids administration in late-adolescence rats. PLOS ONE 7, e31731, http://dx.doi.
org/10.1371/journal.pone.0031731
Aguado, T., Monory, K., Palazuelos, J., Stella, N., Cravatt, B., Lutz, B., Marsicano, G.,
Kokaia, Z., Guzman, M., Galve-Roperh, I., 2005. The endocannabinoid system
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
drives neural progenitor proliferation. FASEB J. 19, 1704–1706, http://dx.doi.
org/10.1096/fj.05-3995fje
Alpár, A., Tortoriello, G., Calvigioni, D., Niphakis, M.J., Milenkovic, I., Bakker, J.,
Cameron, G.A., Hanics, J., Morris, C.V., Fuzik, J., Kovacs, G.G., Cravatt, B.F.,
Parnavelas, J.G., Andrews, W.D., Hurd, Y.L., Keimpema, E., Harkany, T., 2014.
Endocannabinoids modulate cortical development by configuring Slit2/Robo1
signalling. Nat. Commun. 5, 4421, http://dx.doi.org/10.1038/ncomms5421
Antonelli, T., Tomasini, M.C., Tattoli, M., Cassano, T., Tanganelli, S., Finetti, S., Mazzoni,
E., Trabace, L., Steardo, L., Cuomo, V., Ferraro, L., 2005. Prenatal exposure to the
CB1 receptor agonist WIN 55,212-2 causes learning disruption associated with
impaired cortical NMDA receptor function and emotional reactivity changes
in rat offspring. Cereb. Cortex 15, 2013–2020, http://dx.doi.org/10.1093/cercor/
bhi076
Badiani, A., Boden, J.M., De Pirro, S., Fergusson, D.M., Horwood, L.J., Harold, G.T.,
2015. Tobacco smoking and cannabis use in a longitudinal birth cohort: evidence
of reciprocal causal relationships. Drug Alcohol Depend., http://dx.doi.org/10.
1016/j.drugalcdep.2015.02.015
Bambico, F.R., Nguyen, N.T., Katz, N., Gobbi, G., 2010. Chronic exposure to
cannabinoids during adolescence but not during adulthood impairs emotional
behaviour and monoaminergic neurotransmission. Neurobiol. Dis. 37, 641–655,
http://dx.doi.org/10.1016/j.nbd.2009.11.020, S0969-9961(09)00342-8 [pii].
Basavarajappa, B.S., Nagre, N.N., Xie, S., Subbanna, S., 2014. Elevation of endogenous
anandamide impairs LTP, learning, and memory through CB1 receptor signaling
in mice. Hippocampus 24, 808–818, http://dx.doi.org/10.1002/hipo.22272
Batalla, A., Bhattacharyya, S., Yucel, M., Fusar-Poli, P., Crippa, J.A., Nogue, S., Torrens,
M., Pujol, J., Farre, M., Martin-Santos, R., 2013. Structural and functional imaging
studies in chronic cannabis users: a systematic review of adolescent and adult
findings. PLOS ONE 8, e55821, http://dx.doi.org/10.1371/journal.pone.0055821
Battistella, G., Fornari, E., Annoni, J.-M., Chtioui, H., Dao, K., Fabritius, M., Favrat,
B., Mall, J.-F., Maeder, P., Giroud, C., 2014. Long-term effects of cannabis on
brain structure. Neuropsychopharmacology 39, 2041–2048, http://dx.doi.org/
10.1038/npp.2014.67
Behan, A.T., Hryniewiecka, M., O’Tuathaigh, C.M., Kinsella, A., Cannon, M., Karayior-
gou, M., Gogos, J.A., Waddington, J.L., Cotter, D.R., 2012. Chronic adolescent
exposure to delta-9-tetrahydrocannabinol in COMT mutant mice: impact on
indices of dopaminergic, endocannabinoid and GABAergic pathways. Neuropsy-
chopharmacology 37, 1773–1783, http://dx.doi.org/10.1038/npp.2012.24
Belue, R.C., Howlett, A.C., Westlake, T.M., Hutchings, D.E., 1995. The ontogeny of
cannabinoid receptors in the brain of postnatal and aging rats. Neurotoxicol.
Teratol. 17, 25–30.
Benagiano, V., Lorusso, L., Flace, P., Girolamo, F., Rizzi, A., Sabatini, R., Auteri, P.,
Bosco, L., Cagiano, R., Ambrosi, G., 2007. Effects of prenatal exposure to the CB-
1 receptor agonist WIN 55212-2 or CO on the GABAergic neuronal systems of
rat cerebellar cortex. Neuroscience 149, 592–601, http://dx.doi.org/10.1016/j.
neuroscience.2007.07.050
Benes, F.M., 1989. Myelination of cortical-hippocampal relays during late adoles-
cence. Schizophr. Bull. 15, 585–593.
Berghuis, P., Dobszay, M.B., Wang, X., Spano, S., Ledda, F., Sousa, K.M., Schulte, G.,
Ernfors, P., Mackie, K., Paratcha, G., Hurd, Y.L., Harkany, T., 2005. Endocannabi-
noids regulate interneuron migration and morphogenesis by transactivating the
TrkB receptor. Proc. Natl. Acad. Sci. U. S. A. 102, 19115–19120, http://dx.doi.org/
10.1073/pnas.0509494102
Berghuis, P., Rajnicek, A.M., Morozov, Y.M., Ross, R.A., Mulder, J., Urban, G.M.,
Monory, K., Marsicano, G., Matteoli, M., Canty, A., Irving, A.J., Katona, I., Yana-
gawa, Y., Rakic, P., Lutz, B., Mackie, K., Harkany, T., 2007. Hardwiring the brain:
endocannabinoids shape neuronal connectivity. Science 316, 1212–1216, http://
dx.doi.org/10.1126/science.1137406
Berrendero, F., García-Gil, L., Hernández, M.L., Romero, J., Cebeira, M., de
Miguel, R., Ramos, J.A., Fernández-Ruiz, J.J., 1998. Localization of mRNA
expression and activation of signal transduction mechanisms for cannabi-
noid receptor in rat brain during fetal development. Development 125,
3179–3188.
Berrendero, F., Sepe, N., Ramos, J.A., Di Marzo, V., Fernandez-Ruiz, J.J., 1999. Anal-
ysis of cannabinoid receptor binding and mRNA expression and endogenous
cannabinoid contents in the developing rat brain during late gestation and early
postnatal period. Synapse 33, 181–191, http://dx.doi.org/10.1002/(SICI)1098-
2396(19990901)33:3<181::AID-SYN3>3.0.CO;2-R
Biscaia, M., Fernández, B., Higuera-Matas, A., Miguéns, M., Viveros, M.-P.P., García-
Lecumberri, C., Ambrosio, E., Fernandez, B., Miguens, M., Garcia-Lecumberri, C.,
2008. Sex-dependent effects of periadolescent exposure to the cannabinoid ago-
nist CP-55,940 on morphine self-administration behaviour and the endogenous
opioid system. Neuropharmacology 54, 863–873, http://dx.doi.org/10.1016/j.
neuropharm.2008.01.006, S0028-3908(08)00020-8 [pii].
Biscaia, M., Marin, S., Fernandez, B., Marco, E.M., Rubio, M., Guaza, C., Ambrosio,
E., Viveros, M.P., 2003. Chronic treatment with CP 55,940 during the peri-
adolescent period differentially affects the behavioural responses of male and
female rats in adulthood. Psychopharmacology (Berl). 170, 301–308, http://dx.
doi.org/10.1007/s00213-003-1550-7
Blakemore, S.J., 2013. Teenage kicks: cannabis and the adolescent brain.
Lancet 381, 888–889, http://dx.doi.org/10.1016/S0140-6736(12)61578-5,
S0140-6736(12)61578-5 [pii].
Blakemore, S.-J.J., 2012. Imaging brain development: the adolescent brain.
Neuroimage 61, 397–406, http://dx.doi.org/10.1016/j.neuroimage.2011.11.080,
S1053-8119(11)01362-0 [pii].
Bonnin, A., de Miguel, R., Rodriguez-Manzaneque, J.C., Fernandez-Ruiz, J.J., San-
tos, A., Ramos, J.A., 1994. Changes in tyrosine hydroxylase gene expression in
141
mesencephalic catecholaminergic neurons of immature and adult male rats
perinatally exposed to cannabinoids. Brain Res. Dev. Brain Res. 81, 147–150.
Bortolato, M., Bini, V., Frau, R., Devoto, P., Pardu, A., Fan, Y., Solbrig, M.V., 2014. Juve-
nile cannabinoid treatment induces frontostriatal gliogenesis in Lewis rats. Eur.
Neuropsychopharmacol. 24, 974–985, http://dx.doi.org/10.1016/j.euroneuro.
2013.12.011
Bortolato, M., Frau, R., Orrù, M., Casti, A., Aru, G.N., Fà, M., Manunta, M., Usai, A.,
Mereu, G., Gessa, G.L., 2006. Prenatal exposure to a cannabinoid receptor agonist
does not affect sensorimotor gating in rats. Eur. J. Pharmacol. 531, 166–170,
http://dx.doi.org/10.1016/j.ejphar.2005.12.017
Brown, H.L., Graves, C.R., 2013. Smoking and marijuana use in pregnancy.
Clin. Obstet. Gynecol. 56, 107–113, http://dx.doi.org/10.1097/GRF.
0b013e318282377d
Burns, L., Mattick, R.P., Cooke, M., 2006. The use of record linkage to examine illicit
drug use in pregnancy. Addiction 101, 873–882, http://dx.doi.org/10.1111/j.
1360-0443.2006.01444.x
Byrnes, J.J., Johnson, N.L., Schenk, M.E., Byrnes, E.M., 2012. Cannabinoid exposure
in adolescent female rats induces transgenerational effects on morphine condi-
tioned place preference in male offspring. J. Psychopharmacol. 26, 1348–1354,
http://dx.doi.org/10.1177/0269881112443745
Cadoni, C., Simola, N., Espa, E., Fenu, S., Di Chiara, G., 2013. Strain dependence of
adolescent Cannabis influence on heroin reward and mesolimbic dopamine
transmission in adult Lewis and Fischer 344 rats. Addict. Biol., http://dx.doi.
org/10.1111/adb.12085
Calvigioni, D., Hurd, Y.L., Harkany, T., Keimpema, E., 2014. Neuronal substrates and
functional consequences of prenatal cannabis exposure. Eur. Child Adolesc. Psy-
chiatry, http://dx.doi.org/10.1007/s00787-014-0550-y
Campolongo, P., Trezza, V., Cassano, T., Gaetani, S., Morgese, M.G., Ubaldi, M., Sover-
chia, L., Antonelli, T., Ferraro, L., Massi, M., Ciccocioppo, R., Cuomo, V., 2007.
Perinatal exposure to delta-9-tetrahydrocannabinol causes enduring cognitive
deficits associated with alteration of cortical gene expression and neurotrans-
mission in rats. Addict. Biol. 12, 485–495, http://dx.doi.org/10.1111/j.1369-
1600.2007.00074.x
Campolongo, P., Trezza, V., Ratano, P., Palmery, M., Cuomo, V., 2011. Developmental
consequences of perinatal cannabis exposure: behavioral and neuroendocrine
effects in adult rodents. Psychopharmacology (Berl). 214, 5–15, http://dx.doi.
org/10.1007/s00213-010-1892-x
Carli, M., Invernizzi, R.W., 2014. Serotoninergic and dopaminergic modulation of
cortico-striatal circuit in executive and attention deficits induced by NMDA
receptor hypofunction in the 5-choice serial reaction time task. Front. Neural
Circ. 8, 58, http://dx.doi.org/10.3389/fncir.2014.00058
Cass, D.K., Flores-Barrera, E., Thomases, D.R., Vital, W.F., Caballero, A., Tseng, K.Y.,
2014. CB1 cannabinoid receptor stimulation during adolescence impairs the
maturation of GABA function in the adult rat prefrontal cortex. Mol. Psychiatry
19, 536–543, http://dx.doi.org/10.1038/mp.2014.14
Castaldo, P., Magi, S., Cataldi, M., Arcangeli, S., Lariccia, V., Nasti, A.A., Ferraro, L.,
Tomasini, M.C., Antonelli, T., Cassano, T., Cuomo, V., Amoroso, S., 2010. Altered
regulation of glutamate release and decreased functional activity and expression
of GLT1 and GLAST glutamate transporters in the hippocampus of adolescent
rats perinatally exposed to Delta(9)-THC. Pharmacol. Res. 61, 334–341, http://
dx.doi.org/10.1016/j.phrs.2009.11.008, S1043-6618(09)00277-1 [pii].
Castaldo, P., Magi, S., Gaetani, S., Cassano, T., Ferraro, L., Antonelli, T., Amoroso,
S., Cuomo, V., 2007. Prenatal exposure to the cannabinoid receptor agonist
WIN 55,212-2 increases glutamate uptake through overexpression of GLT1 and
EAAC1 glutamate transporter subtypes in rat frontal cerebral cortex. Neurophar-
macology 53, 369–378, http://dx.doi.org/10.1016/j.neuropharm.2007.05.019,
S0028-3908(07)00155-4 [pii].
Castelli, M.P., Paola Piras, A., D’Agostino, A., Pibiri, F., Perra, S., Gessa, G.L., Maccarrone,
M., Pistis, M., 2007. Dysregulation of the endogenous cannabinoid system in
adult rats prenatally treated with the cannabinoid agonist WIN 55,212-2. Eur. J.
Pharmacol. 573, 11–19, http://dx.doi.org/10.1016/j.ejphar.2007.06.047, S0014-
2999(07)00763-7 [pii].
Cha, Y.M., Jones, K.H., Kuhn, C.M., Wilson, W.A., Swartzwelder, H.S., 2007. Sex dif-
ferences in the effects of delta9-tetrahydrocannabinol on spatial learning in
adolescent and adult rats. Behav. Pharmacol. 18, 563–569.
Cha, Y.M., White, A.M., Kuhn, C.M., Wilson, W.a, Swartzwelder, H.S., 2006. Dif-
ferential effects of delta9-THC on learning in adolescent and adult rats.
Pharmacol. Biochem. Behav. 83, 448–455, http://dx.doi.org/10.1016/j.pbb.2006.
03.006, S0091-3057(06)00065-7 [pii].
Chadwick, B., Miller, M.L., Hurd, Y.L., 2013. Cannabis use during adolescent devel-
opment: susceptibility to psychiatric illness. Front. Psychiatry 4, 129, http://dx.
doi.org/10.3389/fpsyt.2013.00129
Chadwick, B., Saylor, A.J., Lopez, H.H., 2011. Adolescent cannabinoid exposure
attenuates adult female sexual motivation but does not alter adulthood CB1R
expression or estrous cyclicity. Pharmacol. Biochem. Behav. 100, 157–164,
http://dx.doi.org/10.1016/j.pbb.2011.07.006, S0091-3057(11)00242-5 [pii].
Chapman, S.L., Wu, L.T., 2013. Substance use among adolescent mothers: a review.
Child Youth Serv. Rev. 35, 806–815, http://dx.doi.org/10.1016/j.childyouth.
2013.02.004
Cornelius, M.D., Leech, S.L., Goldschmidt, L., 2004. Characteristics of persistent smok-
ing among pregnant teenagers followed to young adulthood. Nicotine Tob. Res.
6, 159–169, http://dx.doi.org/10.1080/14622200310001656975
Dalterio, S., Blum, K., DeLallo, L., Sweeney, C., Briggs, A., Bartke, A., 1980.
Perinatal exposure to delta 9-THC in mice: altered enkephalin and nore-
pinephrine sensitivity in vas deferens. Subst. Alcohol Actions. Misuse 1,
467–471.
142 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
Dalterio, S., Steger, R., Mayfield, D., Bartke, A., 1984a. Early cannabinoid exposure
influences neuroendocrine and reproductive functions in male mice: I. Prenatal
exposure. Pharmacol. Biochem. Behav. 20, 107–113.
Dalterio, S., Steger, R., Mayfield, D., Bartke, A., 1984b. Early cannabinoid exposure
influences neuroendocrine and reproductive functions in mice: II. Postnatal
effects. Pharmacol. Biochem. Behav. 20, 115–123.
Day, N.L., Cottreau, C.M., Richardson, G.A., 1993. The epidemiology of alcohol, mari-
juana, and cocaine use among women of childbearing age and pregnant women.
Clin. Obstet. Gynecol. 36, 232–245.
Day, N.L., Goldschmidt, L., Thomas, C.A., 2006. Prenatal marijuana exposure con-
tributes to the prediction of marijuana use at age 14. Addiction 101, 1313–1322,
http://dx.doi.org/10.1111/j.1360-0443.2006.01523.x
Day, N.L., Leech, S.L., Goldschmidt, L., 2011. The effects of prenatal marijuana expo-
sure on delinquent behaviors are mediated by measures of neurocognitive
functioning. Neurotoxicol. Teratol. 33, 129–136, http://dx.doi.org/10.1016/j.ntt.
2010.07.006, S0892-0362(10)00157-1 [pii].
Day, N.L., Richardson, G.A., Geva, D., Robles, N., 1994. Alcohol, marijuana, and
tobacco: effects of prenatal exposure on offspring growth and morphology at
age six. Alcohol. Clin. Exp. Res. 18, 786–794.
Delegación del Gobierno para el Plan Nacional sobre Drogas, 2013. Encuesta
estatal sobre uso de drogas en enseñanzas secundarias (ESTUDES)
2012/2013.
Derauf, C., Kekatpure, M., Neyzi, N., Lester, B., Kosofsky, B., 2009. Neuroimaging of
children following prenatal drug exposure. Semin. Cell Dev. Biol. 20, 441–454,
http://dx.doi.org/10.1016/j.semcdb.2009.03.001, S1084-9521(09)00039-1 [pii].
Díaz-Alonso, J., Aguado, T., de Salas-Quiroga, A., Ortega, Z., Guzmán, M.,
Galve-Roperh, I., 2014. CB1 cannabinoid receptor-dependent activation of
mTORC1/Pax6 signaling drives Tbr2 expression and basal progenitor expansion
in the developing mouse cortex. Cereb. Cortex, http://dx.doi.org/10.1093/cercor/
bhu039
DiNieri, J.A., Wang, X., Szutorisz, H., Spano, S.M., Kaur, J., Casaccia, P., Dow-Edwards,
D., Hurd, Y.L., 2011. Maternal cannabis use alters ventral striatal dopamine D2
gene regulation in the offspring. Biol. Psychiatry 70, 763–769, http://dx.doi.org/
10.1016/j.biopsych.2011.06.027, S0006-3223(11)00672-X [pii].
Duff, G., Argaw, A., Cecyre, B., Cherif, H., Tea, N., Zabouri, N., Casanova, C., Ptito, M.,
Bouchard, J.-F., 2013. Cannabinoid receptor CB2 modulates axon guidance. PLOS
ONE 8, e70849, http://dx.doi.org/10.1371/journal.pone.0070849
Economidou, D., Mattioli, L., Ubaldi, M., Lourdusamy, A., Soverchia, L., Hardiman,
G., Campolongo, P., Cuomo, V., Ciccocioppo, R., 2007. Role of cannabinoider-
gic mechanisms in ethanol self-administration and ethanol seeking in rat adult
offspring following perinatal exposure to Delta9-tetrahydrocannabinol. Toxi-
col. Appl. Pharmacol. 223, 73–85, http://dx.doi.org/10.1016/j.taap.2007.05.008,
S0041-008X(07)00238-4 [pii].
Ehrenreich, H., Rinn, T., Kunert, H.J., Moeller, M.R., Poser, W., Schilling, L.,
Gigerenzer, G., Hoehe, M.R., 1999. Specific attentional dysfunction in adults
following early start of cannabis use. Psychopharmacology (Berl). (142),
295–301.
El Marroun, H., Hudziak, J.J., Tiemeier, H., Creemers, H., Steegers, E.A., Jaddoe, V.W.,
Hofman, A., Verhulst, F.C., van den Brink, W., Huizink, A.C., 2011a. Intrauterine
cannabis exposure leads to more aggressive behavior and attention problems in
18-month-old girls. Drug Alcohol Depend. (118), 470–474, http://dx.doi.org/10.
1016/j.drugalcdep.2011.03.004, S0376-8716(11)00130-X [pii].
El Marroun, H., Jaddoe, V.W., Hudziak, J.J., Roza, S.J., Steegers, E.A., Hofman, A.,
Verhulst, F.C., White, T.J., Stricker, B.H., Tiemeier, H., 2012. Maternal use
of selective serotonin reuptake inhibitors, fetal growth, and risk of adverse
birth outcomes. Arch. Gen. Psychiatry 69, 706–714, http://dx.doi.org/10.1001/
archgenpsychiatry.2011.2333
El Marroun, H., Tiemeier, H., Jaddoe, V.W., Hofman, A., Mackenbach, J.P., Steegers,
E.A., Verhulst, F.C., van den Brink, W., Huizink, A.C., 2008. Demographic,
emotional and social determinants of cannabis use in early pregnancy: the Gen-
eration R study. Drug Alcohol Depend. 98, 218–226, http://dx.doi.org/10.1016/
j.drugalcdep.2008.05.010, S0376-8716(08)00201-9 [pii].
El Marroun, H., Tiemeier, H., Jaddoe, V.W., Hofman, A., Verhulst, F.C., van den Brink,
W., Huizink, A.C., 2011b. Agreement between maternal cannabis use during
pregnancy according to self-report and urinalysis in a population-based cohort:
the Generation R Study. Eur. Addict. Res. 17, 37–43, http://dx.doi.org/10.1159/
000320550
El Marroun, H., Tiemeier, H., Steegers, E.A., Jaddoe, V.W., Hofman, A., Verhulst, F.C.,
van den Brink, W., Huizink, A.C., 2009. Intrauterine cannabis exposure affects
fetal growth trajectories: the Generation R Study. J. Am. Acad. Child Adolesc.
Psychiatry 48, 1173–1181, http://dx.doi.org/10.1097/CHI.0b013e3181bfa8ee,
S0890-8567(09)66073-1 [pii].
El Marroun, H., Tiemeier, H., Steegers, E.A., Roos-Hesselink, J.W., Jaddoe, V.W.,
Hofman, A., Verhulst, F.C., van den Brink, W., Huizink, A.C., 2010. A prospec-
tive study on intrauterine cannabis exposure and fetal blood flow. Early Hum.
Dev. 86, 231–236, http://dx.doi.org/10.1016/j.earlhumdev.2010.03.006, S0378-
3782(10)00079-4 [pii].
Ellgren, M., Artmann, A., Tkalych, O., Gupta, A., Hansen, H.S., Hansen, S.H., Devi,
L.A., Hurd, Y.L., 2008. Dynamic changes of the endogenous cannabinoid and
opioid mesocorticolimbic systems during adolescence: THC effects. Eur. Neu-
ropsychopharmacol. 18, 826–834, http://dx.doi.org/10.1016/j.euroneuro.2008.
06.009, S0924-977X(08)00170-3 [pii].
Ellgren, M., Hurd, Y.L., Franck, J., 2004. Amphetamine effects on dopamine lev-
els and behavior following cannabinoid exposure during adolescence. Eur.
J. Pharmacol. 497, 205–213, http://dx.doi.org/10.1016/j.ejphar.2004.06.048,
S0014-2999(04)00689-2 [pii].
Ellgren, M., Spano, S.M., Hurd, Y.L., 2007. Adolescent cannabis exposure alters opiate
intake and opioid limbic neuronal populations in adult rats. Neuropsychophar-
macology 32, 607–615, http://dx.doi.org/10.1038/sj.npp.1301127
European Monitoring Centre for Drugs and Drug Addiction, 2014. European Drug
Report 2014: Trends and Developments.
Fergusson, D.M., Horwood, L.J., Northstone, K., 2002. Maternal use of cannabis and
pregnancy outcome. BJOG 109, 21–27.
Fontes, M.A., Bolla, K.I., Cunha, P.J., Almeida, P.P., Jungerman, F., Laranjeira, R.R.,
Bressan, R.A., Lacerda, A.L., 2011. Cannabis use before age 15 and subsequent
executive functioning. Br. J. Psychiatry 198, 442–447, http://dx.doi.org/10.1192/
bjp.bp.110.077479
Fried, P., Watkinson, B., James, D., Gray, R., 2002. Current and former marijuana use:
preliminary findings of a longitudinal study of effects on IQ in young adults.
CMAJ 166, 887–891.
Friguls, B., Joya, X., Garcia-Serra, J., Gomez-Culebras, M., Pichini, S., Martinez, S.,
Vall, O., Garcia-Algar, O., 2012. Assessment of exposure to drugs of abuse during
pregnancy by hair analysis in a Mediterranean island. Addiction 107, 1471–1479,
http://dx.doi.org/10.1111/j.1360-0443.2012.03828.x
Gaalema, D.E., Higgins, S.T., Pepin, C.S., Heil, S.H., Bernstein, I.M., 2013. Illicit drug use
among pregnant women enrolled in treatment for cigarette smoking cessation.
Nicotine Tob. Res. 15, 987–991, http://dx.doi.org/10.1093/ntr/nts220
Ganon-Elazar, E., Akirav, I., 2009. Cannabinoid receptor activation in the basolat-
eral amygdala blocks the effects of stress on the conditioning and extinction of
inhibitory avoidance. J. Neurosci. 29, 11078–11088, http://dx.doi.org/10.1523/
JNEUROSCI.1223-09.2009
Garcia, L., de Miguel, R., Ramos, J.A., Fernandez-Ruiz, J.J., 1996. Perinatal delta 9-
tetrahydrocannabinol exposure in rats modifies the responsiveness of midbrain
dopaminergic neurons in adulthood to a variety of challenges with dopaminergic
drugs. Drug Alcohol Depend. 42, 155–166.
Garcia-Gil, L., de Miguel, R., Romero, J., Perez, A., Ramos, J.A., Fernandez-Ruiz, J.J.,
1999. Perinatal delta9-tetrahydrocannabinol exposure augmented the magni-
tude of motor inhibition caused by GABA(B), but not GABA(A), receptor agonists
in adult rats. Neurotoxicol. Teratol. 21, 277–283, S0892036298000580 [pii].
García-Gil, L., Romero, J., Ramos, J.A., Fernández-Ruiz, J.J., 1999. Cannabinoid recep-
tor binding and mRNA levels in several brain regions of adult male and female
rats perinatally exposed to 9-tetrahydrocannabinol. Drug Alcohol Depend. 55,
127–136, http://dx.doi.org/10.1016/S0376-8716(98)00189-6
Giedd, J.N., 2008. The teen brain: insights from neuroimaging. J. Adolesc. Health 42,
335–343, http://dx.doi.org/10.1016/j.jadohealth.2008.01.007
Glass, M., Dragunow, M., Faull, R.L., 1997. Cannabinoid receptors in the human
brain: a detailed anatomical and quantitative autoradiographic study in
the fetal, neonatal and adult human brain. Neuroscience 77, 299–318,
S0306452296004289 [pii].
Gleason, K.A., Birnbaum, S.G., Shukla, A., Ghose, S., 2012. Susceptibility of the ado-
lescent brain to cannabinoids: long-term hippocampal effects and relevance to
schizophrenia. Transl. Psychiatry 2, e199, http://dx.doi.org/10.1038/tp.2012.122
Gomes, F.V., Guimarães, F.S., Grace, A.A., 2014. Effects of pubertal cannabinoid
administration on attentional set-shifting and dopaminergic hyper-responsivity
in a developmental disruption model of schizophrenia. Int. J. Neuropsychophar-
macol. 18, http://dx.doi.org/10.1093/ijnp/pyu018
Gonzalez, B., de Miguel, R., Martin, S., Perez-Rosado, A., Romero, J., Garcia-
Lecumberri, C., Fernandez-Ruiz, J., Ramos, J.A., Ambrosio, E., 2003. Effects of
perinatal exposure to delta 9-tetrahydrocannabinol on operant morphine-
reinforced behavior. Pharmacol. Biochem. Behav. 75, 577–584.
Gonzalez, R., Swanson, J.M., 2012. Long-term effects of adolescent-onset and persis-
tent use of cannabis. Proc. Natl. Acad. Sci. U. S. A. 109, 15970–15971, http://dx.
doi.org/10.1073/pnas.1214124109
Gordon, J.A., Hen, R., 2004. The serotonergic system and anxiety. Neuromolecular
Med. 5, 27–40, http://dx.doi.org/10.1385/NMM:5:1:027
Grant, I., Gonzalez, R., Carey, C.L., Natarajan, L., Wolfson, T., 2003. Non-acute
(residual) neurocognitive effects of cannabis use: a meta-analytic study. J. Int.
Neuropsychol. Soc. 9, 679–689, http://dx.doi.org/10.1017/S1355617703950016
Gruber, S.A., Dahlgren, M.K., Sagar, K.A., Gonenc, A., Killgore, W.D., 2012. Age of
onset of marijuana use impacts inhibitory processing. Neurosci. Lett. 511, 89–94,
http://dx.doi.org/10.1016/j.neulet.2012.01.039, S0304-39401200094-8 [pii].
Harte, L.C., Dow-Edwards, D., 2010. Sexually dimorphic alterations in locomotion
and reversal learning after adolescent tetrahydrocannabinol exposure in the
rat. Neurotoxicol. Teratol. 32, 515–524, http://dx.doi.org/10.1016/j.ntt.2010.05.
001, S0892-0362(10)00109-1 [pii].
Higuera-Matas, A., Botreau, F., Del Olmo, N., Miguens, M., Olias, O., Montoya, G.L.,
Garcia-Lecumberri, C., Ambrosio, E., 2010. Periadolescent exposure to cannabi-
noids alters the striatal and hippocampal dopaminergic system in the adult
rat brain. Eur. Neuropsychopharmacol. 20, 895–906, http://dx.doi.org/10.1016/
j.euroneuro.2010.06.017, S0924-977X(10)00144-6 [pii].
Higuera-Matas, A., Botreau, F., Miguens, M., Del Olmo, N., Borcel, E., Perez-
Alvarez, L., Garcia-Lecumberri, C., Ambrosio, E., Miguéns, M., Pérez-Alvarez,
L., García-Lecumberri, C., 2009. Chronic periadolescent cannabinoid treat-
ment enhances adult hippocampal PSA-NCAM expression in male Wistar rats
but only has marginal effects on anxiety, learning and memory. Pharma-
col. Biochem. Behav. 93, 482–490, http://dx.doi.org/10.1016/j.pbb.2009.06.013,
S0091-3057(09)00209-3 [pii].
Higuera-Matas, A., Miguens, M., Coria, S.M., Assis, M.A., Borcel, E., Del Olmo, N.,
Ambrosio, E., 2012. Sex-specific disturbances of the glutamate/GABA balance
in the hippocampus of adult rats subjected to adolescent cannabinoid exposure.
Neuropharmacology 62, 1975–1984, http://dx.doi.org/10.1016/j.neuropharm.
2011.12.028, S0028-3908(12)00004-4 [pii].
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
Higuera-Matas, A., Soto-Montenegro, M.L., del Olmo, N., Miguens, M., Torres, I.,
Vaquero, J.J., Sanchez, J., Garcia-Lecumberri, C., Desco, M., Ambrosio, E., 2008.
Augmented acquisition of cocaine self-administration and altered brain glucose
metabolism in adult female but not male rats exposed to a cannabinoid agonist
during adolescence. Neuropsychopharmacology 33, 806–813.
Higuera-Matas, A., Soto-Montenegro, M.L., Montoya, G.L., García-Vázquez, V., Pas-
cau, J., Miguéns, M., Del Olmo, N., Vaquero, J.J., García-Lecumberri, C., Desco,
M., Ambrosio, E., 2011. Chronic cannabinoid administration to periadolescent
rats modulates the metabolic response to acute cocaine in the adult brain. Mol.
Imaging Biol. 13, 411–415, http://dx.doi.org/10.1007/s11307-010-0388-8
Howlett, A.C., Barth, F., Bonner, T.I., Cabral, G., Casellas, P., Devane, W.A., Felder,
C.C., Herkenham, M., Mackie, K., Martin, B.R., Mechoulam, R., Pertwee, R.G.,
2002. International Union of Pharmacology. XXVII. Classification of cannabinoid
receptors. Pharmacol. Rev. 54, 161–202.
Huizink, A.C., 2014. Prenatal cannabis exposure and infant outcomes: overview of
studies. Prog. Neuropsychopharmacol. Biol. Psychiatry 52, 45–52, http://dx.doi.
org/10.1016/j.pnpbp.2013.09.014
Hurd, Y.L., Michaelides, M., Miller, M.L., Jutras-Aswad, D., 2014. Trajectory of ado-
lescent cannabis use on addiction vulnerability. Neuropharmacology 76 (Pt B),
416–424, http://dx.doi.org/10.1016/j.neuropharm.2013.07.028
Hurd, Y.L., Wang, X., Anderson, V., Beck, O., Minkoff, H., Dow-Edwards, D., 2005.
Marijuana impairs growth in mid-gestation fetuses. Neurotoxicol. Teratol. 27,
221–229, http://dx.doi.org/10.1016/j.ntt.2004.11.002, S0892-0362(04)00150-3
[pii].
Huttenlocher, P.R., Dabholkar, A.S., 1997. Regional differences in synaptogenesis
in human cerebral cortex. J. Comp. Neurol. 387, 167–178, http://dx.doi.org/10.
1002/(SICI)1096-9861(19971020)387:2<167::AID-CNE1>3.0.CO;2-Z
Jaques, S.C., Kingsbury, A., Henshcke, P., Chomchai, C., Clews, S., Falconer, J., Abdel-
Latif, M.E., Feller, J.M., Oei, J.L., 2014. Cannabis, the pregnant woman and her
child: weeding out the myths. J. Perinatol. 34, 417–424, http://dx.doi.org/10.
1038/jp.2013.180
Jiang, S., Fu, Y., Williams, J., Wood, J., Pandarinathan, L., Avraham, S., Makriyannis, A.,
Avraham, S., Avraham, H.K., 2007. Expression and function of cannabinoid recep-
tors CB1 and CB2 and their cognate cannabinoid ligands in murine embryonic
stem cells. PLoS ONE 2, e641, http://dx.doi.org/10.1371/journal.pone.0000641
Kameda, M., Taylor, C.J., Walker, T.L., Black, D.M., Abraham, W.C., Bartlett, P.F., 2012.
Activation of latent precursors in the hippocampus is dependent on long-term
potentiation. Transl. Psychiatry, http://dx.doi.org/10.1038/tp.2011.70
Kandel, D.B., Yamaguchi, K., Klein, L.C., 2006. Testing the Gateway Hypothesis. Addic-
tion 101, 470–476.
Kawash, G.F., Yeung, D.L., Berg, S.D., 1980. Effects of administration of cannabis resin
during pregnancy on emotionality and learning in rats’ offspring. Percept. Mot.
Skills 50, 359–365.
Keimpema, E., Barabas, K., Morozov, Y.M., Tortoriello, G., Torii, M., Cameron, G.,
Yanagawa, Y., Watanabe, M., Mackie, K., Harkany, T., 2010. Differential sub-
cellular recruitment of monoacylglycerol lipase generates spatial specificity of
2-arachidonoyl glycerol signaling during axonal pathfinding. J. Neurosci. 30,
13992–14007, http://dx.doi.org/10.1523/JNEUROSCI.2126-10.2010
Keimpema, E., Tortoriello, G., Alpár, A., Capsoni, S., Arisi, I., Calvigioni, D., Hu, S.S.-J.,
Cattaneo, A., Doherty, P., Mackie, K., Harkany, T., 2013. Nerve growth factor scales
endocannabinoid signaling by regulating monoacylglycerol lipase turnover in
developing cholinergic neurons. Proc. Natl. Acad. Sci. U. S. A. 110, 1935–1940,
http://dx.doi.org/10.1073/pnas.1212563110
Kim, D., Thayer, S.A., 2001. Cannabinoids inhibit the formation of new synapses
between hippocampal neurons in culture. J. Neurosci. 21, RC146.
Korb, E., Finkbeiner, S., 2011. Arc in synaptic plasticity: from gene to behavior. Trends
Neurosci. 34, 591–598, http://dx.doi.org/10.1016/j.tins.2011.08.007
Korem, N., Akirav, I., 2014. Cannabinoids prevent the effects of a footshock followed
by situational reminders on emotional processing. Neuropsychopharmacology
39, 2709–2722, http://dx.doi.org/10.1038/npp.2014.132
Kosten, T.A., Ambrosio, E., 2002. HPA axis function and drug addictive behaviors:
insights from studies with Lewis and Fischer 344 inbred rats. Psychoneuroen-
docrinology 27, 35–69.
Kowalczyk, T., Pontious, A., Englund, C., Daza, R.A.M., Bedogni, F., Hodge, R., Attardo,
A., Bell, C., Huttner, W.B., Hevner, R.F., 2009. Intermediate neuronal progeni-
tors (basal progenitors) produce pyramidal-projection neurons for all layers of
cerebral cortex. Cereb. Cortex 19, 2439–2450, http://dx.doi.org/10.1093/cercor/
bhn260
Lee, T.T.-Y., Hill, M.N., Hillard, C.J., Gorzalka, B.B., 2013. Temporal changes in
N-acylethanolamine content and metabolism throughout the peri-adolescent
period. Synapse 67, 4–10, http://dx.doi.org/10.1002/syn.21609
Lee, T.T.Y., Wainwright, S.R., Hill, M.N., Galea, L.A.M., Gorzalka, B.B., 2014. Sex,
drugs, and adult neurogenesis: sex-dependent effects of escalating adolescent
cannabinoid exposure on adult hippocampal neurogenesis, stress reactivity,
and amphetamine sensitization. Hippocampus 24, 280–292, http://dx.doi.org/
10.1002/hipo.22221
Lenroot, R.K., Giedd, J.N., 2010. Sex differences in the adolescent brain. Brain Cogn.
72, 46–55, http://dx.doi.org/10.1016/j.bandc.2009.10.008
Lenroot, R.K., Giedd, J.N., 2006. Brain development in children and adolescents:
insights from anatomical magnetic resonance imaging. Neurosci. Biobehav. Rev.
30, 718–729, http://dx.doi.org/10.1016/j.neubiorev.2006.06.001
Lenroot, R.K., Gogtay, N., Greenstein, D.K., Wells, E.M., Wallace, G.L., Clasen, L.S.,
Blumenthal, J.D., Lerch, J., Zijdenbos, A.P., Evans, A.C., Thompson, P.M., Giedd,
J.N., 2007. Sexual dimorphism of brain developmental trajectories during child-
hood and adolescence. Neuroimage 36, 1065–1073, http://dx.doi.org/10.1016/
j.neuroimage.2007.03.053
143
Llorente-Berzal, A., Puighermanal, E., Burokas, A., Ozaita, A., Maldonado, R., Marco,
E.M., Viveros, M.-P., 2013. Sex-dependent psychoneuroendocrine effects of THC
and MDMA in an animal model of adolescent drug consumption. PLOS ONE 8,
e78386, http://dx.doi.org/10.1371/journal.pone.0078386
Long, L.E., Lind, J., Webster, M., Weickert, C.S., 2012. Developmental trajectory of the
endocannabinoid system in human dorsolateral prefrontal cortex. BMC Neu-
rosci. 13, 87, http://dx.doi.org/10.1186/1471-2202-13-87
Lopez-Fernandez, M.A., Montaron, M.F., Varea, E., Rougon, G., Venero, C., Abrous,
D.N., Sandi, C., 2007. Upregulation of polysialylated neural cell adhesion
molecule in the dorsal hippocampus after contextual fear conditioning is
involved in long-term memory formation. J. Neurosci. 27, 4552–4561, http://
dx.doi.org/10.1523/JNEUROSCI.0396-07.2007
López-Gallardo, M., López-Rodríguez, A.B., Llorente-Berzal, Á., Rotllant, D., Mackie,
K., Armario, A., Nadal, R., Viveros, M.-P.P., Lopez-Gallardo, M., Lopez-Rodriguez,
A.B., Llorente-Berzal, A., 2012. Maternal deprivation and adolescent cannabi-
noid exposure impact hippocampal astrocytes, CB1 receptors and brain-derived
neurotrophic factor in a sexually dimorphic fashion. Neuroscience 204, 90–103,
http://dx.doi.org/10.1016/j.neuroscience.2011.09.063, S0306-4522(11)01149-
3 [pii].
Lopez-Rodriguez, A.B., Llorente-Berzal, A., Garcia-Segura, L.M., Viveros, M.-P., 2014.
Sex-dependent long-term effects of adolescent exposure to THC and/or MDMA
on neuroinflammation and serotoninergic and cannabinoid systems in rats. Br.
J. Pharmacol. 171, 1435–1447, http://dx.doi.org/10.1111/bph.12519
Lozano, J., Garcia-Algar, O., Marchei, E., Vall, O., Monleon, T., Giovannandrea, R.D.,
Pichini, S., 2007. Prevalence of gestational exposure to cannabis in a Mediter-
ranean city by meconium analysis. Acta Paediatr. 96, 1734–1737, http://dx.doi.
org/10.1111/j.1651-2227.2007.00535.x
Maccarrone, M., Guzmán, M., Mackie, K., Doherty, P., Harkany, T., 2014. Programming
of neural cells by (endo)cannabinoids: from physiological rules to emerging
therapies. Nat. Rev. Neurosci. 15, 786–801, http://dx.doi.org/10.1038/nrn3846
Maj, P.F., Collu, M., Fadda, P., Cattaneo, A., Racagni, G., Riva, M.A., 2007. Long-
term reduction of brain-derived neurotrophic factor levels and signaling
impairment following prenatal treatment with the cannabinoid receptor 1
receptor agonist (R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinyl-methyl)
pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone. Eur. J.
Neurosci. 25, 3305–3311, http://dx.doi.org/10.1111/j.1460-9568.2007.05565.
x
Marco, E.M., Granstrem, O., Moreno, E., Llorente, R., Adriani, W., Laviola, G., Viveros,
M.P., 2007. Subchronic nicotine exposure in adolescence induces long-term
effects on hippocampal and striatal cannabinoid-CB1 and mu-opioid receptors
in rats. Eur. J. Pharmacol. 557, 37–43, http://dx.doi.org/10.1016/j.ejphar.2006.
11.013, S0014-2999(06)01268-4 [pii].
Marco, E.M., Llorente, R., López-Gallardo, M., Mela, V., Llorente-Berzal, Á., Prada, C.,
Viveros, M.-P., 2015. The maternal deprivation animal model revisited. Neurosci.
Biobehav. Rev. 51, 151–163, http://dx.doi.org/10.1016/j.neubiorev.2015.01.015
Marco, E.M., Rubino, T., Adriani, W., Viveros, M.P., Parolaro, D., Laviola, G., 2009.
Long-term consequences of URB597 administration during adolescence on
cannabinoid CB1 receptor binding in brain areas. Brain Res. 1257, 25–31, http://
dx.doi.org/10.1016/j.brainres.2008.12.037
Mateos, B., Borcel, E., Loriga, R., Luesu, W., Bini, V., Llorente, R., Castelli, M.P., Viveros,
M.P., 2011. Adolescent exposure to nicotine and/or the cannabinoid agonist
CP 55,940 induces gender-dependent long-lasting memory impairments and
changes in brain nicotinic and CB1 cannabinoid receptors. J. Psychopharmacol.
25, 1676–1690, http://dx.doi.org/10.1177/0269881110370503
Mato, S., Del Olmo, E., Pazos, A., 2003. Ontogenetic development of cannabinoid
receptor expression and signal transduction functionality in the human brain.
Eur. J. Neurosci. 17, 1747–1754.
McLaughlin, C.R., Martin, B.R., Compton, D.R., Abood, M.E., 1994. Cannabinoid recep-
tors in developing rats: detection of mRNA and receptor binding. Drug Alcohol
Depend. 36, 27–31.
McPartland, J.M., Glass, M., Pertwee, R.G., 2007. Meta-analysis of cannabinoid lig-
and binding affinity and receptor distribution: interspecies differences. Br. J.
Pharmacol. 152, 583–593, http://dx.doi.org/10.1038/sj.bjp.0707399
McQueeny, T., Padula, C.B., Price, J., Medina, K.L., Logan, P., Tapert, S.F., 2011. Gen-
der effects on amygdala morphometry in adolescent marijuana users. Behav.
Brain Res. 224, 128–134, http://dx.doi.org/10.1016/j.bbr.2011.05.031, S0166-
4328(11)00436-0 [pii].
Meier, M.H., Caspi, A., Ambler, A., Harrington, H., Houts, R., Keefe, R.S.E., McDonald,
K., Ward, A., Poulton, R., Moffitt, T.E., 2012. Persistent cannabis users show neu-
ropsychological decline from childhood to midlife. Proc. Natl. Acad. Sci. U. S. A.
109, E2657–E2664, http://dx.doi.org/10.1073/pnas.1206820109
Mereu, G., Fa, M., Ferraro, L., Cagiano, R., Antonelli, T., Tattoli, M., Ghiglieri, V.,
Tanganelli, S., Gessa, G.L., Cuomo, V., 2003. Prenatal exposure to a cannabi-
noid agonist produces memory deficits linked to dysfunction in hippocampal
long-term potentiation and glutamate release. Proc. Natl. Acad. Sci. U. S. A. 100,
4915–4920, http://dx.doi.org/10.1073/pnas.0537849100
Mitsuhiro, S.S., Chalem, E., Barros, M.C., Guinsburg, R., Laranjeira, R., 2007. Prevalence
of cocaine and marijuana use in the last trimester of adolescent preg-
nancy: socio-demographic, psychosocial and behavioral characteristics. Addict.
Behav. 32, 392–397, http://dx.doi.org/10.1016/j.addbeh.2006.05.001, S0306-
4603(06)00151-1 [pii].
Molina-Holgado, F., Amaro, A., Gonzalez, M.I., Alvarez, F.J., Leret, M.L., 1996. Effect
of maternal delta 9-tetrahydrocannabinol on developing serotonergic system.
Eur. J. Pharmacol. 316, 39–42.
Molina-Holgado, F., Molina-Holgado, E., Leret, M.L., González, M.I., Reader, T.A.,
1993. Distribution of indoleamines and [3H]paroxetine binding in rat brain
144 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
regions following acute or perinatal delta 9-tetrahydrocannabinol treatments.
Neurochem. Res. 18, 1183–1191.
Moore, D.G., Turner, J.D., Parrott, A.C., Goodwin, J.E., Fulton, S.E., Min, M.O., Fox, H.C.,
Braddick, F.M., Axelsson, E.L., Lynch, S., Ribeiro, H., Frostick, C.J., Singer, L.T., 2010.
During pregnancy, recreational drug-using women stop taking ecstasy (3,4-
methylenedioxy-N-methylamphetamine) and reduce alcohol consumption, but
continue to smoke tobacco and cannabis: initial findings from the Develop-
ment and Infancy Study. J. Psychopharmacol. 24, 1403–1410, http://dx.doi.org/
10.1177/0269881109348165
Moore, H., Jentsch, J.D., Ghajarnia, M., Geyer, M.A., Grace, A.A., 2006. A neurobehav-
ioral systems analysis of adult rats exposed to methylazoxymethanol acetate on
E17: implications for the neuropathology of schizophrenia. Biol. Psychiatry 60,
253–264, http://dx.doi.org/10.1016/j.biopsych.2006.01.003
Morel, L.J., Giros, B., Dauge, V., 2009. Adolescent exposure to chronic delta-
9-tetrahydrocannabinol blocks opiate dependence in maternally deprived
rats. Neuropsychopharmacology 34, 2469–2476, http://dx.doi.org/10.1038/npp.
2009.70
Moreno, M., Trigo, J.M., Escuredo, L., Rodriguez de Fonseca, F., Navarro, M., 2003. Peri-
natal exposure to delta 9-tetrahydrocannabinol increases presynaptic dopamine
D2 receptor sensitivity: a behavioral study in rats. Pharmacol. Biochem. Behav.
75, 565–575, S0091305703001175 [pii].
Morishita, J., Okamoto, Y., Tsuboi, K., Ueno, M., Sakamoto, H., Maekawa, N.,
Ueda, N., 2005. Regional distribution and age-dependent expression of N-
acylphosphatidylethanolamine-hydrolyzing phospholipase D in rat brain. J.
Neurochem. 94, 753–762, http://dx.doi.org/10.1111/j.1471-4159.2005.03234.
x S0028-3908(10)00224-8 [pii].
Morris, C.V., DiNieri, J.A., Szutorisz, H., Hurd, Y.L., 2011. Molecular mechanisms of
maternal cannabis and cigarette use on human neurodevelopment. Eur. J. Neu-
rosci. 34, 1574–1583, http://dx.doi.org/10.1111/j.1460-9568.2011.07884.x
Mulder, J., Aguado, T., Keimpema, E., Barabas, K., Ballester Rosado, C.J., Nguyen, L.,
Monory, K., Marsicano, G., Di Marzo, V., Hurd, Y.L., Guillemot, F., Mackie, K., Lutz,
B., Guzman, M., Lu, H.C., Galve-Roperh, I., Harkany, T., 2008. Endocannabinoid
signaling controls pyramidal cell specification and long-range axon patterning.
Proc. Natl. Acad. Sci. U. S. A. 105, 8760–8765, http://dx.doi.org/10.1073/pnas.
0803545105
Nacher, J., Guirado, R., Castillo-Gómez, E., 2013. Structural plasticity of interneurons
in the adult brain: role of PSA-NCAM and implications for psychiatric disorders.
Neurochem. Res. 38, 1122–1133, http://dx.doi.org/10.1007/s11064-013-0977-
4 Rivkin, M.J., Davis, P.E., Lemaster, J.L., Cabral, H.J., Warfield, S.K., Mulkern, R.V., Rob-
Newsom, R.J., Kelly, S.J., 2008. Perinatal delta-9-tetrahydrocannabinol exposure dis-
rupts social and open field behavior in adult male rats. Neurotoxicol. Teratol. 30,
213–219, http://dx.doi.org/10.1016/j.ntt.2007.12.007, S0892-0362(07)00367-4
[pii].
Noland, J.S., Singer, L.T., Short, E.J., Minnes, S., Arendt, R.E., Kirchner, H.L., Bearer,
C., 2005. Prenatal drug exposure and selective attention in preschoolers.
Neurotoxicol. Teratol. 27, 429–438, http://dx.doi.org/10.1016/j.ntt.2005.02.001,
S0892-0362(05)00040-1 [pii].
O’Shea, M., McGregor, I.S., Mallet, P.E., 2006. Repeated cannabinoid exposure during
perinatal, adolescent or early adult ages produces similar longlasting deficits in
object recognition and reduced social interaction in rats. J. Psychopharmacol.
20, 611–621.
O’Shea, M., Singh, M.E., McGregor, I.S., Mallet, P.E., 2004. Chronic cannabi-
noid exposure produces lasting memory impairment and increased
anxiety in adolescent but not adult rats. J. Psychopharmacol. 18,
502–508.
O’Tuathaigh, C.M.P., Hryniewiecka, M., Behan, A., Tighe, O., Coughlan, C., Desbonnet,
L., Cannon, M., Karayiorgou, M., Gogos, J.A., Cotter, D.R., Waddington, J.L., 2010.
Chronic adolescent exposure to -9-tetrahydrocannabinol in COMT mutant
mice: impact on psychosis-related and other phenotypes. Neuropsychophar-
macology 35, 2262–2273, http://dx.doi.org/10.1038/npp.2010.100
Oudin, M.J., Gajendra, S., Williams, G., Hobbs, C., Lalli, G., Doherty, P., 2011. Endo-
cannabinoids regulate the migration of subventricular zone-derived neuroblasts
in the postnatal brain. J. Neurosci. 31, 4000–4011, http://dx.doi.org/10.1523/
jneurosci.5483-10.2011
Padula, C.B., Schweinsburg, A.D., Tapert, S.F., 2007. Spatial working memory perfor-
mance and fMRI activation interaction in abstinent adolescent marijuana users.
Psychol. Addict. Behav. 21, 478–487, http://dx.doi.org/10.1037/0893-164X.21.
4.478
Paria, B.C., Dey, S.K., 2000. Ligand-receptor signaling with endocannabinoids in
preimplantation embryo development and implantation. Chem. Phys. Lipids
108, 211–220, S0009308400001973 [pii].
Perez-Rosado, A., Gomez, M., Manzanares, J., Ramos, J.A., Fernandez-Ruiz, J., 2002.
Changes in prodynorphin and POMC gene expression in several brain regions of
rat fetuses prenatally exposed to Delta(9)-tetrahydrocannabinol. Neurotoxicol.
Res. 4, 211–218, http://dx.doi.org/10.1080/10298420290023936
Perez-Rosado, A., Manzanares, J., Fernandez-Ruiz, J., Ramos, J.A., 2000. Prenatal
Delta(9)-tetrahydrocannabinol exposure modifies proenkephalin gene expres-
sion in the fetal rat brain: sex-dependent differences. Brain Res. Dev. Brain Res.
120, 77–81.
Pertwee, R.G., 2008. The diverse CB1 and CB2 receptor pharmacology of three
plant cannabinoids: delta9-tetrahydrocannabinol, cannabidiol and delta9-
tetrahydrocannabivarin. Br. J. Pharmacol. 153, 199–215, http://dx.doi.org/10.
1038/sj.bjp.0707442
Petit-Demouliere, B., Chenu, F., Bourin, M., 2005. Forced swimming test in mice:
a review of antidepressant activity. Psychopharmacology (Berl). 177, 245–255,
http://dx.doi.org/10.1007/s00213-004-2048-7
Pistis, M., Perra, S., Pillolla, G., Melis, M., Muntoni, A.L., Gessa, G.L., 2004. Adolescent
exposure to cannabinoids induces long-lasting changes in the response to drugs
of abuse of rat midbrain dopamine neurons. Biol. Psychiatry 56, 86–94.
Pope Jr., H.G., Gruber, A.J., Hudson, J.I., Huestis, M.A., Yurgelun-Todd, D., 2001. Neu-
ropsychological performance in long-term cannabis users. Arch. Gen. Psychiatry
58, 909–915, yoa20391 [pii].
Quinn, H.R., Matsumoto, I., Callaghan, P.D., Long, L.E., Arnold, J.C., Gunasekaran, N.,
Thompson, M.R., Dawson, B., Mallet, P.E., Kashem, M.A., Matsuda-Matsumoto,
H., Iwazaki, T., McGregor, I.S., 2008. Adolescent rats find repeated Delta(9)-
THC less aversive than adult rats but display greater residual cognitive deficits
and changes in hippocampal protein expression following exposure. Neuropsy-
chopharmacology 33, 1113–1126.
Raver, S.M., Keller, A., 2014. Permanent suppression of cortical oscillations in mice
after adolescent exposure to cannabinoids: receptor mechanisms. Neurophar-
macology 86, 161–173, http://dx.doi.org/10.1016/j.neuropharm.2014.07.006
Raznahan, A., Shaw, P.W., Lerch, J.P., Clasen, L.S., Greenstein, D., Berman, R., Pipitone,
J., Chakravarty, M.M., Giedd, J.N., 2014. Longitudinal four-dimensional mapping
of subcortical anatomy in human development. Proc. Natl. Acad. Sci. U. S. A. 111,
1592–1597, http://dx.doi.org/10.1073/pnas.1316911111
Realini, N., Rubino, T., Parolaro, D., 2009. Neurobiological alterations at adult age
triggered by adolescent exposure to cannabinoids. Pharmacol. Res. 60, 132–138.
Realini, N., Vigano, D., Guidali, C., Zamberletti, E., Rubino, T., Parolaro, D.,
2011. Chronic URB597 treatment at adulthood reverted most depressive-like
symptoms induced by adolescent exposure to THC in female rats. Neurophar-
macology 60, 235–243, http://dx.doi.org/10.1016/j.neuropharm.2010.09.003,
Reimold, M., Batra, A., Knobel, A., Smolka, M.N., Zimmer, A., Mann, K., Solbach, C.,
Reischl, G., Schwärzler, F., Gründer, G., Machulla, H.-J., Bares, R., Heinz, A., 2008.
Anxiety is associated with reduced central serotonin transporter availability in
unmedicated patients with unipolar major depression: a [11C]DASB PET study.
Mol. Psychiatry 13, 606–613, http://dx.doi.org/10.1038/sj.mp.4002149, 557.
Renard, J., Krebs, M., Le Pen, G., Jay, T.M., 2014. Long-term consequences of adoles-
cent cannabinoid exposure in adult psychopathology. Front. Neurosci. 8, 1–14,
http://dx.doi.org/10.3389/fnins.2014.00361
Renard, J., Krebs, M.O., Jay, T.M., Le Pen, G., 2013. Long-term cognitive impair-
ments induced by chronic cannabinoid exposure during adolescence in rats:
a strain comparison. Psychopharmacology (Berl). 225, 781–790, http://dx.doi.
org/10.1007/s00213-012-2865-z
son, C.D., Rose-Jacobs, R., Frank, D.A., 2008. Volumetric MRI study of brain in
children with intrauterine exposure to cocaine, alcohol, tobacco, and marijuana.
Pediatrics 121, 741–750, http://dx.doi.org/10.1542/peds.2007-1399
Robbins, T.W., 2007. Shifting and stopping: fronto-striatal substrates, neurochemical
modulation and clinical implications. Philos. Trans. R. Soc. Lond. Ser. B: Biol. Sci.
362, 917–932, http://dx.doi.org/10.1098/rstb.2007.2097, H01V0404612T2871
[pii].
Rodriguez de Fonseca, F., Ramos, J.A., Bonnin, A., Fernandez-Ruiz, J.J., 1993. Presence
of cannabinoid binding sites in the brain from early postnatal ages. Neuroreport
4, 135–138.
Romero, J., Garcia-Palomero, E., Berrendero, F., Garcia-Gil, L., Hernandez, M.L.,
Ramos, J.A., Fernandez-Ruiz, J.J., 1997. Atypical location of cannabinoid receptors
in white matter areas during rat brain development. Synapse 26, 317–323.
Ronn, L.C., Berezin, V., Bock, E., 2000. The neural cell adhesion molecule in synaptic
plasticity and ageing. Int. J. Dev. Neurosci. 18, 193–199.
Rubino, T., Parolaro, D., 2008. Long lasting consequences of cannabis exposure in
adolescence. Mol. Cell. Endocrinol. 286, S108–S113.
Rubino, T., Prini, P., Piscitelli, F., Zamberletti, E., Trusel, M., Melis, M., Sagheddu, C.,
Ligresti, A., Tonini, R., Di Marzo, V., Parolaro, D., 2014. Adolescent exposure to
THC in female rats disrupts developmental changes in the prefrontal cortex.
Neurobiol. Dis., http://dx.doi.org/10.1016/j.nbd.2014.09.015 (in press).
Rubino, T., Realini, N., Braida, D., Guidi, S., Capurro, V., Vigano, D., Guidali, C., Pinter,
M., Sala, M., Bartesaghi, R., Parolaro, D., 2009. Changes in hippocampal morphol-
ogy and neuroplasticity induced by adolescent THC treatment are associated
with cognitive impairment in adulthood. Hippocampus 19, 763–772.
Rubino, T., Vigano’, D., Realini, N., Guidali, C., Braida, D., Capurro, V., Castiglioni,
C., Cherubino, F., Romualdi, P., Candeletti, S., Sala, M., Parolaro, D., Vigano,
D., 2008. Chronic Delta(9)-tetrahydrocannabinol during adolescence provokes
sex-dependent changes in the emotional profile in adult rats: behavioral and
biochemical correlates. Neuropsychopharmacology 33, 2760–2771, http://dx.
doi.org/10.1038/sj.npp.1301664
Rubio, P., Rodriguez de Fonseca, F., Munoz, R.M., Ariznavarreta, C., Martin-Calderon,
J.L., Navarro, M., 1995. Long-term behavioral effects of perinatal exposure to
delta 9-tetrahydrocannabinol in rats: possible role of pituitary-adrenal axis. Life
Sci. 56, 2169–2176.
Saez, T.M.M., Aronne, M.P., Caltana, L., Brusco, A.H., 2014. Prenatal exposure to the
CB1 and CB2 cannabinoid receptor agonist WIN 55,212-2 alters migration of
early-born glutamatergic neurons and GABAergic interneurons in the rat cere-
bral cortex. J. Neurochem. 129, 637–648, http://dx.doi.org/10.1111/jnc.12636
Saurel-Cubizolles, M.-J., Prunet, C., Blondel, B., 2014. Cannabis use during pregnancy
in France in 2010. BJOG 121, 971–977, http://dx.doi.org/10.1111/1471-0528.
12626
Schneider, M., 2009. Cannabis use in pregnancy and early life and its consequences:
animal models. Eur. Arch. Psychiatry Clin. Neurosci. 259, 383–393, http://dx.doi.
org/10.1007/s00406-009-0026-0
Schneider, M., 2008. Puberty as a highly vulnerable developmental period for the
consequences of cannabis exposure. Addict. Biol. 13, 253–263.
 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
Schneider, M., Drews, E., Koch, M., 2005. Behavioral effects in adult rats of chronic
prepubertal treatment with the cannabinoid receptor agonist WIN 55,212-2.
Behav. Pharmacol. 16, 447–454.
Schneider, M., Koch, M., 2003. Chronic pubertal, but not adult chronic cannabinoid
treatment impairs sensorimotor gating, recognition memory, and the perfor-
mance in a progressive ratio task in adult rats. Neuropsychopharmacology 28,
1760–1769.
Schulz, S., Becker, T., Nagel, U., von Ameln-Mayerhofer, A., Koch, M., 2013. Chronic
co-administration of the cannabinoid receptor agonist WIN55,212-2 during
puberty or adulthood reverses 3,4 methylenedioxymetamphetamine (MDMA)-
induced deficits in recognition memory but not in effort-based decision making.
Pharmacol. Biochem. Behav. 106, 91–100, http://dx.doi.org/10.1016/j.pbb.2013.
03.011, S0091-3057(13)00077-4 [pii].
Schweinsburg, A.D., Schweinsburg, B.C., Medina, K.L., McQueeny, T., Brown, S.A.,
Tapert, S.F., 2010. The influence of recency of use on fMRI response during spa-
tial working memory in adolescent marijuana users. J. Psychoactive Drugs 42,
401–412.
Segev, A., Rubin, A.S., Abush, H., Richter-Levin, G., Akirav, I., 2014. Cannabinoid recep-
tor activation prevents the effects of chronic mild stress on emotional learning
and LTP in a rat model of depression. Neuropsychopharmacology 39, 919–933,
http://dx.doi.org/10.1038/npp.2013.292
Shabani, M., Hosseinmardi, N., Haghani, M., Shaibani, V., Janahmadi, M., 2011. Mater-
nal exposure to the CB1 cannabinoid agonist WIN 55212-2 produces robust
changes in motor function and intrinsic electrophysiological properties of cere-
bellar Purkinje neurons in rat offspring. Neuroscience 172, 139–152, http://dx.
doi.org/10.1016/j.neuroscience.2010.10.031, S0306-4522(10)01361-8 [pii].
Shabani, M., Mahnam, A., Sheibani, V., Janahmadi, M., 2014. Alterations in the intrin-
sic burst activity of Purkinje neurons in offspring maternally exposed to the CB1
cannabinoid agonist WIN 55212-2. J. Membr. Biol. 247, 63–72, http://dx.doi.org/
10.1007/s00232-013-9612-1
Silva, L., Zhao, N., Popp, S., Dow-Edwards, D., 2012. Prenatal tetrahydrocannabinol
(THC) alters cognitive function and amphetamine response from weaning to
adulthood in the rat. Neurotoxicol. Teratol. 34, 63–71, http://dx.doi.org/10.1016/
j.ntt.2011.10.006, S0892-0362(11)00205-4 [pii].
Smith, A.M., Fried, P.A., Hogan, M.J., Cameron, I., 2006. Effects of prenatal mar-
ijuana on visuospatial working memory: an fMRI study in young adults.
Neurotoxicol. Teratol. 28, 286–295, http://dx.doi.org/10.1016/j.ntt.2005.12.008,
S0892-0362(05)00180-7 [pii].
Smith, A.M., Fried, P.A., Hogan, M.J., Cameron, I., 2004. Effects of prenatal marijuana
on response inhibition: an fMRI study of young adults. Neurotoxicol. Teratol.
26, 533–542, http://dx.doi.org/10.1016/j.ntt.2004.04.004, S0892036204000662
[pii].
Smith, M.J., Cobia, D.J., Reilly, J.L., Gilman, J.M., Roberts, A.G., Alpert, K.I., Wang, L., Bre-
iter, H.C., Csernansky, J.G., 2015. Cannabis-related episodic memory deficits and
hippocampal morphological differences in healthy individuals and schizophre-
nia subjects. Hippocampus, http://dx.doi.org/10.1002/hipo.22427
Spano, M.S., Ellgren, M., Wang, X., Hurd, Y.L., 2007. Prenatal cannabis exposure
increases heroin seeking with allostatic changes in limbic enkephalin systems
in adulthood. Biol. Psychiatry 61, 554–563, http://dx.doi.org/10.1016/j.biopsych.
2006.03.073, S0006-3223(06)00557-9 [pii].
Stopponi, S., Soverchia, L., Ubaldi, M., Cippitelli, A., Serpelloni, G., Ciccocioppo, R.,
2014. Chronic THC during adolescence increases the vulnerability to stress-
induced relapse to heroin seeking in adult rats. Eur. Neuropsychopharmacol.
24, 1037–1045, http://dx.doi.org/10.1016/j.euroneuro.2013.12.012
Suarez, I., Bodega, G., Fernandez-Ruiz, J., Ramos, J.A., Rubio, M., Fernandez, B., 2004a.
Down-regulation of the AMPA glutamate receptor subunits GluR1 and GluR2/3
in the rat cerebellum following pre- and perinatal delta9-tetrahydrocannabinol
exposure. Cerebellum 3, 66–74.
Suarez, I., Bodega, G., Fernandez-Ruiz, J.J., Ramos, J.A., Rubio, M., Fernandez, B., 2002.
Reduced glial fibrillary acidic protein and glutamine synthetase expression in
astrocytes and Bergmann glial cells in the rat cerebellum caused by delta(9)-
tetrahydrocannabinol administration during development. Dev. Neurosci. 24,
300–312.
Suarez, I., Bodega, G., Rubio, M., Fernandez-Ruiz, J.J., Ramos, J.A., Fernandez, B.,
2004b. Prenatal cannabinoid exposure down-regulates glutamate transporter
expressions (GLAST and EAAC1) in the rat cerebellum. Dev. Neurosci. 26,
45–53.
Suarez, J., Llorente, R., Romero-Zerbo, S.Y., Mateos, B., Bermudez-Silva, F.J., de
Fonseca, F.R., Viveros, M.P., 2009. Early maternal deprivation induces gender-
dependent changes on the expression of hippocampal CB(1) and CB(2)
cannabinoid receptors of neonatal rats. Hippocampus 19, 623–632, http://dx.
doi.org/10.1002/hipo.20537
Substance Abuse and Mental Health Services Administration, 2014a. Results from
the 2013 National Survey on Drug Use and Health: Summary of National Find-
ings.
Substance Abuse and Mental Health Services Administration, 2014b. Behavioral
Health Barometer.
Substance Abuse and Mental Health Services Administration, 2013. Results from the
2012 National Survey on Drug Use and Health.
Substance Abuse and Mental Health Services Administration, 2009. Substance Use
among Women During Pregnancy and Following Childbirth, The NSDUH Report.
Sun, X., Dey, S.K., 2008. Aspects of endocannabinoid signaling in periimplanta-
tion biology. Mol. Cell. Endocrinol. 286, S3–S11, http://dx.doi.org/10.1016/j.mce.
2008.01.002, S0303-7207(08)00008-7 [pii].
Szutorisz, H., DiNieri, J.a, Sweet, E., Egervari, G., Michaelides, M., Carter, J.M., Ren,
Y., Miller, M.L., Blitzer, R.D., Hurd, Y.L., 2014. Parental THC exposure leads to
145
compulsive heroin-seeking and altered striatal synaptic plasticity in the sub-
sequent generation. Neuropsychopharmacology 39, 1315–1323, http://dx.doi.
org/10.1038/npp.2013.352
Tantra, M., Kröcher, T., Papiol, S., Winkler, D., Röckle, I., Jatho, J., Burkhardt, H., Ron-
nenberg, A., Gerardy-Schahn, R., Ehrenreich, H., Hildebrandt, H., 2014. St8sia2
deficiency plus juvenile cannabis exposure in mice synergistically affect higher
cognition in adulthood. Behav. Brain Res., http://dx.doi.org/10.1016/j.bbr.2014.
08.062
Tapert, S.F., Schweinsburg, A.D., Drummond, S.P.A., Paulus, M.P., Brown, S.A., Yang,
T.T., Frank, L.R., 2007. Functional MRI of inhibitory processing in abstinent ado-
lescent marijuana users. Psychopharmacology (Berl). 194, 173–183, http://dx.
doi.org/10.1007/s00213-007-0823-y
Thanos, P.K., Michaelides, M., Umegaki, H., Volkow, N.D., 2008. D2R DNA transfer into
the nucleus accumbens attenuates cocaine self-administration in rats. Synapse
62, 481–486, http://dx.doi.org/10.1002/syn.20523
Tomasiewicz, H.C., Jacobs, M.M., Wilkinson, M.B., Wilson, S.P., Nestler, E.J.,
Hurd, Y.L., 2012. Proenkephalin mediates the enduring effects of adolescent
cannabis exposure associated with adult opiate vulnerability. Biol. Psychi-
atry 72, 803–810, http://dx.doi.org/10.1016/j.biopsych.2012.04.026, S0006-
3223(12)00409-X [pii].
Tortoriello, G., Morris, C.V., Alpar, A., Fuzik, J., Shirran, S.L., Calvigioni, D., Keimpema,
E., Botting, C.H., Reinecke, K., Herdegen, T., Courtney, M., Hurd, Y.L., Harkany,
T., 2014. Miswiring the brain: 9-tetrahydrocannabinol disrupts cortical devel-
opment by inducing an SCG10/stathmin-2 degradation pathway. EMBO J. 33,
668–685, http://dx.doi.org/10.1002/embj.201386035
Tree, K.C., Scotto di Perretolo, M., Peyronnet, J., Cayetanot, F., 2014. In utero cannabi-
noid exposure alters breathing and the response to hypoxia in newborn mice.
Eur. J. Neurosci. 40, 2196–2204, http://dx.doi.org/10.1111/ejn.12588
Trezza, V., Campolongo, P., Cassano, T., Macheda, T., Dipasquale, P., Carratu,
M.R., Gaetani, S., Cuomo, V., 2008. Effects of perinatal exposure to delta-9-
tetrahydrocannabinol on the emotional reactivity of the offspring: a longitudinal
behavioral study in Wistar rats. Psychopharmacology (Berl). 198, 529–537,
http://dx.doi.org/10.1007/s00213-008-1162-3
Trezza, V., Campolongo, P., Manduca, A., Morena, M., Palmery, M., Vanderschuren,
L.J., Cuomo, V., 2012. Altering endocannabinoid neurotransmission at critical
developmental ages: impact on rodent emotionality and cognitive performance.
Front. Behav. Neurosci. 6, 2, http://dx.doi.org/10.3389/fnbeh.2012.00002
U.S. Department of Health and Human Services, 1999. Blending perspective and
building common ground. A report to Congress on substance abuse and
child protection [WWW Document]. URL: http://aspe.hhs.gov/hsp/subabuse99/
subabuse.htm (accessed 10.06.14).
United Nations Office on Drugs and Crime, 2013. World Drug Report 2013. Malta.
Van Zundert, B., Yoshii, A., Constantine-Paton, M., 2004. Receptor compartmental-
ization and trafficking at glutamate synapses: a developmental proposal. Trends
Neurosci. 27, 428–437, http://dx.doi.org/10.1016/j.tins.2004.05.010, S0166-
2236(04)00164-X [pii].
Vassoler, F.M., Johnson, N.L., Byrnes, E.M., 2013. Female adolescent exposure to
cannabinoids causes transgenerational effects on morphine sensitization in
female offspring in the absence of in utero exposure. J. Psychopharmacol. 27,
1015–1022, http://dx.doi.org/10.1177/0269881113503504
Vela, G., Fuentes, J.A., Bonnin, A., Fernandez-Ruiz, J., Ruiz-Gayo, M., 1995.
Perinatal exposure to delta 9-tetrahydrocannabinol (delta 9-THC) leads
to changes in opioid-related behavioral patterns in rats. Brain Res. 680,
142–147.
Vela, G., Martin, S., Garcia-Gil, L., Crespo, J.A., Ruiz-Gayo, M., Javier Fernandez-
Ruiz, J., Garcia-Lecumberri, C., Pelaprat, D., Fuentes, J.A., Ramos, J.A., Ambrosio,
E., 1998. Maternal exposure to delta9-tetrahydrocannabinol facilitates mor-
phine self-administration behavior and changes regional binding to central mu
opioid receptors in adult offspring female rats. Brain Res. 807, 101–109, S0006-
8993(98)00766-5 [pii].
Venero, C., Herrero, A.I., Touyarot, K., Cambon, K., Lopez-Fernandez, M.A., Berezin,
V., Bock, E., Sandi, C., 2006. Hippocampal up-regulation of NCAM expression
and polysialylation plays a key role on spatial memory. Eur. J. Neurosci. 23,
1585–1595.
Viveros, M.P., Llorente, R., Suarez, J., Llorente-Berzal, A., Lopez-Gallardo, M., de
Fonseca, F.R., Rodriguez de Fonseca, F., 2011a. The endocannabinoid system
in critical neurodevelopmental periods: sex differences and neuropsychi-
atric implications. J. Psychopharmacol. 26, 164–176, http://dx.doi.org/10.1177/
0269881111408956
Viveros, M.P., Marco, E.M., Lopez-Gallardo, M., Garcia-Segura, L.M., Wagner, E.J.,
2011b. Framework for sex differences in adolescent neurobiology: a focus on
cannabinoids. Neurosci. Biobehav. Rev. 35, 1740–1751, http://dx.doi.org/10.
1016/j.neubiorev.2010.09.005, S0149-7634(10)00145-4 [pii].
Volkow, N.D., Fowler, J.S., Wang, G.J., Swanson, J.M., 2004. Dopamine in
drug abuse and addiction: results from imaging studies and treatment
implications. Mol. Psychiatry 9, 557–569, http://dx.doi.org/10.1038/sj.mp.
4001507 4001507
Walters, D.E., Carr, L.A., 1988. Perinatal exposure to cannabinoids alters neuro-
chemical development in rat brain. Pharmacol. Biochem. Behav. 29, 213–216,
S0091-3057(88)90300-0 [pii].
Wang, X., Dow-Edwards, D., Anderson, V., Minkoff, H., Hurd, Y.L., 2006. Discrete
opioid gene expression impairment in the human fetal brain associated with
maternal marijuana use. Pharmacogenomics J. 6, 255–264, http://dx.doi.org/10.
1038/sj.tpj.6500375
Wang, X., Dow-Edwards, D., Anderson, V., Minkoff, H., Hurd, Y.L., 2004. In utero
marijuana exposure associated with abnormal amygdala dopamine D2 gene
146 A. Higuera-Matas et al. / Neuroscience and Biobehavioral Reviews 55 (2015) 119–146
expression in the human fetus. Biol. Psychiatry 56, 909–915, http://dx.doi.org/
10.1016/j.biopsych.2004.10.015, S0006-3223(04)01095-9 [pii].
Wang, X., Dow-Edwards, D., Keller, E., Hurd, Y.L., 2003. Preferential limbic expression
of the cannabinoid receptor mRNA in the human fetal brain. Neuroscience 118,
681–694, S0306452203000204 [pii].
Warner, T.D., Behnke, M., Eyler, F.D., Padgett, K., Leonard, C., Hou, W., Garvan, C.W.,
Schmalfuss, I.M., Blackband, S.J., 2006. Diffusion tensor imaging of frontal white
matter and executive functioning in cocaine-exposed children. Pediatrics 118,
2014–2024, http://dx.doi.org/10.1542/peds.2006-0003
Wegener, N., Koch, M., 2009. Behavioural disturbances and altered Fos protein
expression in adult rats after chronic pubertal cannabinoid treatment. Brain
Res. 1253, 81–91, http://dx.doi.org/10.1016/j.brainres.2008.11.081, S0006-
8993(08)02848-5 [pii].
Wenger, T., Gerendai, I., Fezza, F., Gonzalez, S., Bisogno, T., Fernandez-Ruiz, J., Di
Marzo, V., 2002. The hypothalamic levels of the endocannabinoid, anandamide,
peak immediately before the onset of puberty in female rats. Life Sci. 70,
1407–1414.
Wu, C.S., Jew, C.P., Lu, H.C., 2011. Lasting impacts of prenatal cannabis exposure and
the role of endogenous cannabinoids in the developing brain. Futur. Neurol. 6,
459–480.
Wu, C.-S., Morgan, D., Jew, C.P., Andrews, M.-J., Leishman, E., Spencer, C.M., Czyzyk,
T., Bradshaw, H., Mackie, K., Lu, H.-C., 2014. Long-term consequences of perinatal
fatty acid amino hydrolase inhibition. Br. J. Pharmacol. 171, 1420–1434.
Wu, C.S., Zhu, J., Wager-Miller, J., Wang, S., O’Leary, D., Monory, K., Lutz, B., Mackie, K.,
Lu, H.C., 2010. Requirement of cannabinoid CB(1) receptors in cortical pyramidal
neurons for appropriate development of corticothalamic and thalamocorti-
cal projections. Eur. J. Neurosci. 32, 693–706, http://dx.doi.org/10.1111/j.1460-
9568.2010.07337.x
Yamamoto, K., Shinba, T., Yoshii, M., 2014. Psychiatric symptoms of noradrener-
gic dysfunction: a pathophysiological view. Psychiatry Clin. Neurosci. 68, 1–20,
http://dx.doi.org/10.1111/pcn.12126
Yan, H.C., Cao, X., Das, M., Zhu, X.H., Gao, T.M., 2010. Behavioral animal models of
depression. Neurosci. Bull. 26, 327–337, http://dx.doi.org/10.1007/s12264-010-
0323-7
Zamberletti, E., Beggiato, S., Steardo, L., Prini, P., Antonelli, T., Ferraro, L., Rubino,
T., Parolaro, D., 2014. Alterations of prefrontal cortex GABAergic transmission
in the complex psychotic-like phenotype induced by adolescent delta-9-
tetrahydrocannabinol exposure in rats. Neurobiol. Dis. 63, 35–47, http://dx.doi.
org/10.1016/j.nbd.2013.10.028
Zamberletti, E., Prini, P., Speziali, S., Gabaglio, M., Solinas, M., Parolaro, D., Rubino,
T., 2012. Gender-dependent behavioral and biochemical effects of adolescent
delta-9-tetrahydrocannabinol in adult maternally deprived rats. Neuroscience
204, 245–257, http://dx.doi.org/10.1016/j.neuroscience.2011.11.038, S0306-
4522(11)01323-6 [pii].
Zhang, Y., Landthaler, M., Schlussman, S.D., Yuferov, V., Ho, A., Tuschl, T., Kreek, M.J.,
2009. Mu opioid receptor knockdown in the substantia nigra/ventral tegmental
area by synthetic small interfering RNA blocks the rewarding and locomo-
tor effects of heroin. Neuroscience 158, 474–483, http://dx.doi.org/10.1016/j.
neuroscience.2008.09.039
Zhu, P.J., Lovinger, D.M., 2010. Developmental alteration of endocannabinoid retro-
grade signaling in the hippocampus. J. Neurophysiol. 103, 1123–1129, http://dx.
doi.org/10.1152/jn.00327.2009
Zurolo, E., Iyer, A.M., Spliet, W.G., Van Rijen, P.C., Troost, D., Gorter, J.A., Aronica, E.,
2010. CB1 and CB2 cannabinoid receptor expression during development and in
epileptogenic developmental pathologies. Neuroscience 170, 28–41, http://dx.
doi.org/10.1016/j.neuroscience.2010.07.004, S0306-4522(10)00958-9 [pii].