Oxytocin and microglia in the
development of social behaviour
royalsocietypublishing.org/journal/rstb
Review
Cite this article: Gonzalez A, Hammock EAD.
2022 Oxytocin and microglia in the
development of social behaviour. Phil.
Trans. R. Soc. B 377: 20210059.
Downloaded from https://royalsocietypublishing.org/ on 26 October 2022
https://doi.org/10.1098/rstb.2021.0059
Received: 2 February 2022
Accepted: 18 April 2022
One contribution of 15 to a theme issue
‘Interplays between oxytocin and other
neuromodulators in shaping complex social
behaviours’.
Subject Areas:
neuroscience, developmental biology,
behaviour
Keywords:
oxytocin, microglia, development, social
behaviour, sex differences
Author for correspondence:
Elizabeth A. D. Hammock
e-mail: hammock@psy.fsu.edu
Alicia Gonzalez and Elizabeth A. D. Hammock
Department of Psychology and Program in Neuroscience, Florida State University, 1107 West Call Street,
Tallahassee, FL 32306, USA
AG, 0000-0002-1495-1208; EADH, 0000-0001-7668-745X
Oxytocin is a well-established regulator of social behaviour. Microglia, the
resident immune cells of the central nervous system, regulate brain develop-
ment and maintenance in health and disease. Oxytocin and microglia
interact: microglia appear to regulate the oxytocin system and are, in turn,
regulated by oxytocin, which appears to have anti-inflammatory effects.
Both microglia and oxytocin are regulated in sex-specific ways. Oxytocin
and microglia may work together to promote experience-dependent circuit
refinement through multiple developmental-sensitive periods contributing
to individual differences in social behaviour.
This article is part of the theme issue ‘Interplays between oxytocin and
other neuromodulators in shaping complex social behaviours’.
1. Introduction
Advances in understanding, diagnosing and reporting social behaviour
disorders, such as autism spectrum disorder (ASD), are associated with
increased diagnostic prevalence globally. According to the Centers for Disease
Control and Prevention, 1.85% of US children aged 8 were diagnosed with ASD
in 2016. Additionally, a UK multi-centre study estimated the prevalence of ASD
to be 1.57% [1] in 2009. Recent standardized epidemiological assessment of
ASD in China [2] and Qatar [3] have reported similar prevalence. Other dis-
orders, like anxiety and depression, which can be improved or made worse
by the quality of available social interaction, have a global prevalence of 7.3%
[4] and 5% according to the World Health Organization. Anxiety- and depress-
ive-like behaviours are also markedly increased in mouse models [5] of ASD
and in humans [6] diagnosed with ASD. Given the prevalence of these diag-
noses, researchers are keen to identify neural and environmental mechanisms
that ameliorate or exacerbate challenges associated with them. Oxytocin regu-
lates social behaviours such as attachment and partner preference [7–9],
including a potential role in ASD. Interestingly, emerging evidence has
suggested microglia may also play a role in social behaviour development,
with a potential contribution of dysregulated microglia in the aetiology of
ASD [10,11]. More recently, it has been suggested that oxytocin and microglia
systems interact during neurodevelopment, with implications for neurodeve-
lopmental disorders like ASD [12–18]. In this review, we focus on the
available evidence for microglia to regulate the development of the oxytocin
system and for oxytocin to exert an anti-inflammatory role on microglia, and
their contribution to social behaviour development in health and disease.
2. Duality of microglia
Microglia, the innate immune cells of the central nervous system (CNS), work to
maintain a homeostatic environment [19]. Comprising 5–10% of the total
number of brain cells [20], microglia interact with other cells of the brain
throughout life, playing critical roles in development, learning and brain main-
tenance, and in response to injury or pathogens. Microglia have a spectrum of
phenotypic presentations with an amoeboid ‘activated’ phenotype thought to
© 2022 The Author(s) Published by the Royal Society. All rights reserved.
 (a) 2
microglia
ramified amoeboid

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(b)
concentration

neurodegeneration
chemokine

CX3CL1
ADP
IL-6 neuroprotection
synaptic homeostasis IL-1b
TNFa
healthy vulnerable
Figure 1. Microglia display a dynamic morphology and expression of chemokines and cytokines throughout their interactions with neurons. (a) Microglia (top row)
vary in their morphology from ramified (left) to amoeboid (right). Ramified microglia play an active role in synapse regulation between healthy neurons. Amoeboid
microglia are associated with neuronal vulnerability due to tissue damage and/or infection which activate microglia. (b) While in the ramified state, microglia release
chemokines and cytokines at low levels to maintain a homeostatic synaptic environment. As microglia become activated and transition into the amoeboid mor-
phology as a result of infection or injury, they release more chemokines and cytokines and contribute to the vulnerability of the neuron. A rapid return to lower
levels of inflammatory chemokines and cytokines is associated with improved outcomes, while chronic elevation of inflammatory chemokines and cytokines after
microglia activation is associated with poor neural outcomes. Created with BioRender.com.

play a role in immune functions and ramified ‘resting’ pheno-
type playing a role in critical synaptic regulatory functions
(figure 1). While they provide these important functions to
regulate brain health, microglia can also become dysregulated
with sustained activation [21,22] and contribute to neuro-
pathology [23,24].
Adult microglia maintain a steady state turn-over rate of
renewal and elimination that varies by brain region [25–27].
Unlike circulating macrophages which have a mesodermal
origin, most microglia are of endodermal origin and reside
and renew within the brain. However, it is possible that
during sickness, injury or disease, inflammatory circulating
monocytes may enter the brain parenchyma and differentiate
into cells with microglia-like phenotypes [28,29]. Whether
they are resident microglia or invade from blood in severe
injury or disease, inflammatory microglia are amoeboid in
shape and secrete inflammatory cytokines including
tumour necrosis factor α (TNFα), interleukin-1α (IL-1α) and
interleukin-1β (IL-1β) [30,31]. It is upon introduction of a
pathogen or debris that these cells enter into an active state.
A classical and simplified view of activated microglia
argued that there are two extreme ends of a gradient of acti-
vated microglia, M1 and M2. In this simplified dichotomy,
the polarization (M1 or M2) of their activation depends on
their downstream response. That is, M1 microglia character-
istically release pro-inflammatory cytokines, while M2
microglia release anti-inflammatory cytokines. It is important
to note that this polarization is not necessarily binary. More
recent data demonstrate more nuance and diversity of micro-
glia function. Though the terms M1 and M2 no longer fully
encompass the phenotypic diversity of microglia, the diver-
sity of microglia function (i.e. similar or even opposing
functions) is crucial in maintaining a homeostatic environ-
ment [32,33] and to help regulate the crucial balance of
cytokine release [34]. Microglia express signal detecting cell
surface receptors such as toll-like receptors (TLRs), Fc recep-
tors, CD36 and RAGE [35–38], all of which assist in
responding to the local environment. As resident macro-
phages in the brain parenchyma patrolling the spaces
between neurons, microglia use phagocytosis to clean up
injured cells or pathogens [39]. Because neurons are generally
non-mitotic, the rapid clearing of invading pathogens or
accumulation of cellular debris is an important component
of the overall function of microglia. In neurogenic areas of
the adult brain, microglia play a role in the engulfment of
neuronal precursor cells, thus controlling the pool of poten-
tial neurons in adults [40].
More than just phagocytosis in injury and disease, micro-
glia help to regulate many aspects of healthy brain function
as they monitor brain activity at the synaptic and cellular
level [41–43], playing an important role in learning and
memory. In their ‘resting’ ramified state, microglia survey
synapses. Well-studied hippocampal circuits that use N-
methyl-D-aspartate receptor activation for long-term poten-
tiation in synaptic plasticity appear to depend on signals
from microglia [44–46]. Microglia also play a role in synaptic
scaling through TNFα signalling, which is proposed to occur
during sleep to change relative synaptic strength to maintain
informative synapses while also leaving room for new synap-
tic plasticity [47–50]. Microglia scan the brain parenchyma
 and make contact with active synapses [41–43]. In zebrafish
time-lapse video recordings, the frequency of synapse scan-
ning activity is positively correlated with highly active
synapses. Once microglia have scanned (made direct contact
with) an active synapse, synaptic activity is reduced [51],
suggesting that this scanning contact from microglia was
the cause of the reduction in synaptic activity.
In short, microglia have characteristic responses that are
both necessary for and harmful to healthy brain function
[29,41,52–55]. Whether patrolling the brain parenchyma for
synaptic activity or serving their neuroimmune functions,
microglia use the same cytokines and chemokines and
phagocytic processes (figure 1). This duality of microglial
function raises fundamental questions of how brains may
be at special risk during development when microglia are
both setting up brain architecture and potentially negotiating
neuroinflammatory insults throughout critical periods for
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circuit formation and sensitive periods for experience-
dependent circuit refinement. The duality of microglia is
made more complex by the presence of numerous robust
sex differences in activity and response to inflammatory
events at different developmental stages, putting males and
females at unequal risk for different adverse outcomes [56].
3. Microglia in development
Derived from primitive macrophages within the blood
islands of the embryonic yolk sac [57], microglia are present
early in brain development with roles in neurogenesis
[58,59] and apoptosis [60–62], synaptogenesis and develop-
mental synaptic pruning [63–65], and wiring [66–68].
Microglia contribute to critical early developmental processes
like facilitating the travel of numerous axons into a nerve
bundle (fasciculation) [69] and ensuring vascular cell tip
fusion of developing blood vessels [70], as well as later devel-
opmental processes wherein lived experience shapes brain
development.
The primitive macrophages are believed to migrate to
the developing brain parenchyma through the develop-
ing vasculature [39], ventricular layer [71–74] and the
meninges through the pial surface [52,71–73,75–77]. The
brain parenchyma lures microglia through these routes
using chemoattractants, such as colony-stimulating factor 1
(CSF-1) [57,78], interleukin-34 and matrix metalloproteinases
(MMP-8 and MMP-9) [79]. Once recruited to the brain par-
enchyma, microglia can use structural support and
directional guidance of radial glia to migrate to different
areas or cortical layers, where they can be functionally inte-
grated with cells native to the CNS [72,80–85]. The brain
parenchyma also produces instructive signals, including
TGFβ [86] and other unidentified factors, for the maturation
of microglia, accomplished during the first two postnatal
weeks in mice [87]. The first postnatal week is marked by
high microglia proliferation rates. During proliferation,
these cells adopt an amoeboid morphology and exhibit
increased phagocytosis. As the cells mature, extensive
branches protrude from the soma [88]. By the end of the
second postnatal week, microglia display a mature adult-
like ramified phenotype, often referred to as ‘resting’ micro-
glia [87]—a name which seems out of touch with modern
live imaging data that show that these ramified cells are
very active [41]. While the cells themselves mature, their
population peaks in number in the first two postnatal 3
weeks and starts to decline in population such that by six
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weeks of age, the population of mature ramified microglia
is about 50% of their developmental peak [89]. Also, while
resting ramified microglia are the predominant microglial
morph, there are other forms determined by the local
environment in the brain parenchyma [29,52,90–96].
Microglia participate in the developmental regulation of
synaptic pruning [63,64,97]. Early in development during
synaptogenesis, the brain overproduces synapses. Microglia
contribute to the process of synapse elimination
[42,61,63,65,67], with impacts on social behaviour in adult-
hood [98]. First, microglia tag the weak synapses, then
engulf them [63,64,97]. Mice without the ability to tag weak
Phil. Trans. R. Soc. B 377: 20210059
synapses (C1q knockout mice) display seizures and higher
than typical numbers of excitatory synapses [65,99]. In a
mouse model of optic nerve injury, the expression of comp-
lement C1q, C3 and C3 receptor (CR3) mRNA and protein
were upregulated following injury. CR3+ microglia phagocy-
tose C1q opsonized myelin debris after optic nerve injury
[100]. Increased engulfment of synapses by microglia also
occurs with an overexpression of C4a, resulting in a reduction
of synaptic density in the cortex and altered social behaviour
in adulthood [98]. CX3CR1 knockout mice have impaired
microglia production and overabundant synapses in the
CA1 hippocampal region during development. These mice
also have a behavioural phenotype that includes impairments
in social behaviour [65,101]. Microglia play a role in experi-
ence-dependent plasticity of numerous brain systems,
including the visual system [42,43], with an experience-expec-
tant critical period in mice from 21–35 postnatal days [102].
For example, mice without P2RY12—another important
regulator of microglia—have deficits in synaptic plasticity
in the visual cortex [103].
Iba1 is a well-established marker of microglia, used
frequently to identify microglia for cell counting and mor-
phological analysis. Mice with a conventional global
deletion of the gene encoding this protein, Aif1, demonstrate
marked impairments in neurodevelopmental events as well
as behaviour in adulthood [104]. Using an alternative
marker to identify microglia, loss of Iba1 did not impair the
numbers of microglia in the developing brain, but the loss
of Iba1 did impair the function of microglia. Iba1-deficient
mice showed dysregulated expression of synaptic pruning
markers (some were elevated, some were reduced). Hippo-
campal CA1 microglia in Iba1-deficient mice were less
ramified, and the neural activity of excitatory neurons was
diminished. As adults, Iba1-deficient mice were tested in a
variety of tasks (open field, elevated plus maze, novel
object recognition and sociability). The Iba1-deficient mice
showed increased locomotor behaviour, no change in
anxiety-like components of the behavioural tasks, impaired
novel object recognition and impaired sociability. Like the
prior studies, this provides clear evidence for an important
contribution of microglia in the regulation of brain function
and associated behaviour, beginning in development.
4. Neuroinflammation
While microglia play important roles in development, micro-
glia also contribute to immune reactivity of the brain and
contribute to neuroinflammation with potential for impairing
 (a) 4

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bacteria
virus
PRR
PGDH
IL-6
IL-1b
TNFa

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(b) (c)
extracellular space
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TLR4 IL1R IL6R OXTR

BBB

NF-kB NF-IL6
cytoplasm

IL-6
nucleus

NF-kB NF-IL6
OXTR

Figure 2. Oxytocin receptor (OXTR) expression changes in response to immune mediators within the fetal environment. (a) Placental macrophages express PRRs and
recognize surfactant protein, bacteria and viruses. Once recognized, downstream signalling cascades result in the release of immune mediators necessary for parturi-
tion, but harmful to the fetus if overexpressed. (b) Increased peripheral cytokines infiltrate the CNS and activate microglia. As microglial cells become activated, OXTR
expression on the cell membrane increases. The deactivation of microglial cells is OXTR-mediated. (c) In this simplified intracellular signalling cascade, various
immune mediators contribute to OXTR membrane expression. Solid arrows represent known pathways, while the dashed arrow represents a hypothesized pathway
based on known kinase activation of NF-IL-6 [111]. The phosphorylation of NF-κB [112] and NF-IL-6 [111] by downstream signalling of TLR4, IL-1R, IL-6R and OXTR
promote translocation into the nucleus, where they act as transcription factors in a synergistic manner on the OXTR gene. Created with BioRender.com.

brain function [10]. Inflammation is characterized by the acti-
vation of immune cells in response to harmful stimuli, which
include pathogens, damaged cells and irritants. The mount-
ing of an immune response serves to clear the damage and
initiate repair. As the first line of defence, the innate
immune system throughout the organism attempts to stave
off noxious stimuli using a variety of physical and chemical
barriers, such as the skin and mucosa [105]. Additionally,
effector cells like the epithelial cell layers that make up the
skin, endothelial cells expressing cell adhesion molecules
(CAMs) and cadherins, or monocytes that secrete pathogen
degrading enzymes and phagocytose, are all components of
innate immunity [106]. This mechanism of host defence is
evolutionarily ancient, and as such uses biochemical signa-
tures known as pathogen-associated molecular patterns
(PAMPs) found in a broad spectrum of bacterial and fungal
cell walls to distinguish invaders from host cells [107]. The
PAMPs are recognized by pattern recognition receptors
(PRRs) which include four classes—TLRs, C-type lectin
receptors, RIG-I-like receptors and nucleotide-binding oligo-
merization domain [107]. Additionally, PRRs are activated
by molecules that are released from damaged or dying cells
known as damage-associated molecular patterns.
The initiation of innate effector cells is rapid and prompts
the onset of the adaptive immune system, which is composed
of some innate effector cells but additionally recruits lympho-
cytes. Adaptive immunity is important in the formation of
immunological memory of specific pathogens, allowing for
better host defence upon reinfection [108] (reviewed in [82]).
Though the mature CNS is protected from peripheral
pathogen exposure due to the presence of the blood–brain-
barrier, disruption of this barrier in adulthood, or its imma-
ture state in development, can lead to peripheral infiltration
and subsequent central immune activation. Upregulation of
peripheral cytokines, specifically IL-6, induces the expression
of vascular cell adhesion molecule 1 (VCAM-1) on endo-
thelial cells [109]. VCAM-1 mediates the adhesion and
transmigration of leucocytes as well as IL-6 [110] (figure 2).
As peripheral infiltration continues, increasing concentration
of IL-6 activates T-cells which subsequently shed their IL-6
receptors (IL-6R) via metalloproteinase-mediated proteolytic
cleavage, specifically by ADAM17 [113]. The shed IL-6Rs
bind the neighbouring IL-6 to coordinate trans-signalling
and subsequent activation of microglia and astrocytes [114].
Fetal microglia activation has been linked to social develop-
mental disorders, like ASD. Using histochemical techniques,
brain samples of individuals diagnosed with ASD showed
abnormal microglial morphology associated with the activated
phenotype and increased pro-inflammatory cytokine profiles
[115–117]. Dysregulation of the oxytocin system has been
 causally linked to social behaviour disruption in animal models
and has been associated with ASD in humans [118,119].
Though the direct interplay between oxytocin and microglia
in ASD has not yet been studied, oxytocin and microglia inter-
actions have been explored in numerous other contexts.
Microglia and oxytocin appear to regulate each other, with
developmental age and sex influencing this potential for co-
regulation.
5. Microglia and oxytocin
(a) Microglia regulate the oxytocin system
One of the first studies identifying microglia as a potential
modulator of oxytocin used immunohistochemical tech-
niques along with electron microscopy to reveal that
microglial cells make up about 19% of the neurohypophysis,
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where they make contact with and engulf magnocellular
terminals, suggesting their ability to modulate the release of
oxytocin into the periphery [120]. In a drug-induced model
of diabetes mellitus in adult male rats, microglia appeared
to phagocytose oxytocin neurons of the paraventricular
nucleus (PVN) of the hypothalamus and their terminals in
the posterior pituitary [121]. Additionally, injections of IL-1β
(a cytokine released by microglia) into both the periphery
[122] and the brain [123–125] activate oxytocin neurons.
Peripheral injury such as gut perforation can activate
microglia in the brain. In turn, these activated microglia can
modulate oxytocin release into the periphery [126]. Cecal lig-
ation and puncture (CLP) is an animal model of sepsis due to
perforated intestines. Adult male Wistar rats were used to
evaluate what happens to plasma levels of hormones after
the CLP procedure with and without microglia function.
After CLP, Iba1 protein levels (a marker of microglia) in the
brain were elevated at 6 and 24 h. A microglia inhibitor, min-
ocycline, was delivered directly to the brain to evaluate the
impact of microglia inhibition on circulating hormones.
Microglia inhibition reduced the increase in Iba1 seen after
CLP. Brains were collected for protein quantification by wes-
tern blot, and ELISAs were used to measure hormones in
blood plasma. Within 6 h of CLP, circulating oxytocin levels
were nearly doubled. Microglia inhibition with minocycline
prevented this increase [126]. Combined, this evidence indi-
cates that the intestinal injury activated microglia leading to
an oxytocin response in adults.
Microglia regulate not only oxytocin, but also potentially
the oxytocin receptor (OXTR), through their release of cyto-
kines. The OXTR gene encodes a 7-transmembrane protein
that is coupled with a Gq/11α class G-protein [127]. Upon
activation of OXTR via ligand binding, phospholipase C-β
is stimulated resulting in the enzymatic cleavage of phospha-
tidylinositol biphosphate (PIP2) into inositol trisphosphate
(IP3) and 1,2-diacylglycerol (DAG). The binding of IP3 to
receptors expressed on the endoplasmic reticulum triggers
calcium release from internal stores. The increase in intra-
cellular calcium has a variety of effects on specific tissue
types. Brain expression patterns of OXTR are dynamic
throughout life [128].
OXTR levels are regulated by the pro-inflammatory cyto-
kine, interleukin-6 (IL-6), as established in a variety of cell
types. Because microglia are a major producer of IL-6
within the brain, it is hypothesized that some of the dynamic
regulation of OXTR across time and brain areas might be
related to IL-6 released from microglia. Within the 17 kilobase 5
human OXTR gene lies two transcription start sites 618 and
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621 base pairs upstream of the methionine initiation codon
[129]. Binding sites for several transcription factors were
found near these start sites, including one for nuclear factor
interleukin-6 (NF-IL-6) [129]. The addition of IL-6 and IL-1β
has been shown to translocate NF-IL-6 into the nucleus and
regulate OXTR expression, though some conflicting evidence
concerning the direction of regulation has been published.
One study using immortalized human myometrial cells trea-
ted with IL-6 and IL-1β resulted in a decrease of OXTR
mRNA expression [130]. By contrast, another study obtained
biopsied myometrial tissue from women during elective
cesarean delivery, in which isolated cells treated with IL-1β
Phil. Trans. R. Soc. B 377: 20210059
increased OXTR mRNA expression within 4 h [131]. Further,
the increased activity of NF-IL-6 upregulated OXTR mRNA
expression in response to mechanical stretch of human uter-
ine myocytes [132].
However, the NF-IL-6 binding site is not the only one
within the OXTR gene regulatory region that responds to
local cytokines. A binding site for nuclear factor kappa B
(NF-κB) was also identified within the regulatory region of
the OXTR gene. NF-κB translocation is induced by cytokines,
hormones, transmitters and growth factors. The binding of
NF-κB to its DNA binding site spurs transcription of genes
involved in cell survival, positive and negative feedback,
and inflammation [133–135]. NF-κB works mutually with
NF-IL-6 to finely upregulate OXTR expression in myometrial
cells isolated from humans [131]. In primary macrophage
cells isolated from humans and mice, NF-κB activation regu-
lates OXTR [136]. OXTR mRNA and protein expression was
upregulated when treated with lipopolysaccharide (LPS).
However, the increased expression of OXTR was hindered
upon NF-κB inhibition, suggesting OXTR expression is regu-
lated via NF-κB [136].
In addition to pro-inflammatory-related binding sequences,
the OXTR promoter region contains partial estrogen response
elements (ERE) that may contribute to the sex-specific
expression of OXTR [137]. Two classes of nuclear estrogen
receptors bind to the ERE subsequently promoting transcrip-
tion, estrogen receptor alpha and beta (ERα, ERβ—
respectively) [138–140]. Regulation of OXTR mRNA expression
via oestradiol in human uterine cells has been shown to be con-
centration dependent [141]. Experiments using a murine model
identified similar regulation of OXTR in that expression is
modulated by the addition of oestradiol in the uterus, but
also in the pituitary and brain [142–144]. The increase in
OXTR expression is mediated by the activation of ERα, while
activation of ERβ is responsible for the maintenance of basal
OXTR expression [145]. Interestingly, ERβ is the only estrogen
receptor class that is expressed by microglia, demonstrated
using an autoimmune encephalomyelitis mouse model [146].
Activation of microglial ERβ suppresses the TLR4-mediated
inflammatory response by potentially acting as a transcriptional
co-repressor as it interacts with the repressor C-terminal-bind-
ing protein (CtBP) [147].
These studies show a process of peripheral injury or
stimulation leading to microglia activation followed by
altered oxytocin function, with the potential for feedback
regulation via cytokine regulation of the OXTR. From these
studies alone, it is not clear what role oxytocin might play,
but in combination with the studies below, it appears that
oxytocin may serve in a negative feedback role to help
 reign in microglia and prevent or reduce inflammatory
damage.
(b) Oxytocin regulates microglia
Oxytocin regulates peripheral macrophages [148–150]
through OXTR activation [136,151,152] and appears to simi-
larly regulate microglia. Cultured human vascular cells and
macrophages express OXTR mRNA. Cells treated with phys-
iological concentrations of oxytocin exhibited a decrease in
superoxide activity and IL-6 secretion [153]. The addition of
oxytocin blunted the action of LPS, an endotoxin, and
decreased the expression of pro-inflammatory cytokines
such as IL-1β, IL-6 and TNFα. Moreover, oxytocin increased
response to IL-4 and stimulated the production of genes
characteristic of the anti-inflammatory macrophagic pheno-
type, including arginase I (Arg1) [152]. Oxytocin inhibits
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NF-κB signalling and alleviates LPS-induced stress in
Caco2BB enterocytes [154]. The infusion of oxytocin into
obese mice reduced the size of adipocytes and decreased
levels of the cytokines IL-6 and TNFα and the systemic
inflammatory marker amyloid A [148]. Thus, there is ample
evidence that oxytocin activity in the periphery is associated
with a reduction in inflammatory activity.
In the brain, manipulation of the oxytocin system impacts
microglia function or markers of activity, although it is less
clear if microglia express OXTRs in vivo. An early indication
that oxytocin might regulate microglia came from an inciden-
tal finding using the microdialysis technique to evaluate
extracellular contents in the brain. Microdialysis probes infus-
ing an acidic solution adjacent to the oxytocin-rich supraoptic
nucleus were used in adult male rats and after 6 h, a signifi-
cant microglia response (as indicated by hypertrophied OX-
42 immunoreactive cells) was noted [155]. This inflammatory
response was prevented with a subcutaneous injection of
hypertonic saline [123], a known inducer of oxytocin release
from PVN magnocellular cells [156].
Direct evidence of oxytocin regulation of microglia is
apparent from cell culture studies. It is important to note
that recent RNA sequencing efforts suggest little-to-no Oxtr
expression in microglia of healthy postnatal mice in vivo
[87,157]. However, in cell culture, microglia express OXTR
[158,159]. These conflicting results might be explained by
alterations in microglia transcriptome due to serum exposure,
or when not in the presence of factors secreted by the CNS
environment [160]. Despite this important confound, cell cul-
ture remains an important tool to evaluate direct effects of
oxytocin on cellular processes. Inflammatory responses can
be experimentally induced with LPS which binds to a PRR,
TLR4 [161], in the cell membrane of macrophages and micro-
glia. Oxytocin given to microglia in the cell culture media
reduced the levels of LPS-induced activation of microglia
[158]. Pretreatment of oxytocin followed by LPS exposure in
primary microglial cells cultured from the cortex of neonatal
mice or cells from the BV-2 microglial cell line significantly
reduced the activation state of both cell types compared to
cells treated only with LPS. Additionally, the expression of
pro-inflammatory markers (TNFα and IL-1β) was reduced
in the oxytocin-pretreated group. Oxytocin had no effect on
these markers in the absence of LPS. OXTR mRNA and
protein expression was elevated upon LPS exposure [159].
A similar anti-inflammatory role for oxytocin has been docu-
mented in an additional microglia cell line (MG6) [162].
Oxytocin pretreatment appears anti-inflammatory in vivo 6
as well. LPS injected into the intraperitoneal cavity of adult
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male mice induced a standard inflammatory response in
the prefrontal cortex with increased microglia activation
(amoeboid shape and more intense Iba1 staining), which
was significantly reduced by pretreatment with intranasal
oxytocin. Oxytocin did not affect these parameters in the
absence of LPS treatment. The mRNA and protein for
TNFα and IL-1β were both increased by LPS treatment,
which was ameliorated by oxytocin pretreatment. LPS
increased TNFα in microglia (Iba1 + cells) and astrocytes
(GFAP+); however, oxytocin only blunted the microglia
response to LPS [159]. Combined, these data provide evidence
that oxytocin may act on microglia cells to apply the brakes to
Phil. Trans. R. Soc. B 377: 20210059
pro-inflammatory processes within the brain. In this manner,
oxytocin acting within the brain could serve a neuroprotective
role during neuroinflammation across the lifespan.
Oxytocin may serve as an anti-inflammatory not just for
the innate immune response to endotoxins like LPS, but
also to other insults like traumatic brain injury. Like oxytocin,
treatment with a non-steroidal anti-inflammatory drug, salsa-
late, reduced levels of TNFα and IL-1β in response to LPS in
BV-2 cells. In vivo, experimental traumatic brain injury induced
Iba1 immunofluorescence in adult mice, but this effect was
blunted with salsalate, coincident with the decreased expression
of some inflammatory response genes, and a NF-κB gene set,
and upregulated neurogenesis and neuroprotection-associated
transcripts including oxytocin and vasopressin [163]. Perhaps
salsalate is effective at dampening pro-inflammatory cas-
cades after injury through its upregulation of oxytocin.
Oxytocin levels are regulated in part by social experience.
For example, co-housing with an ovariectomized female for a
period of one week increased oxytocin mRNA production by
about 40% in adult male mice compared to mice housed
alone during that time period [158]. There is strong clinical
evidence that social support helps individuals recover from
stroke [164]. Using a middle cerebral artery occlusion stroke
model, socially housed male mice had decreased infarct
size after occlusion compared to mice housed alone. Isolated
males treated with oxytocin had reduced infarct size, but not
if the oxytocin administration was combined with an OXTR
antagonist. Socially housed mice treated with a selective
OXTR antagonist had larger infarct size similar to the isolate
housed males. Combined, these data suggest that social hous-
ing increases oxytocin signalling through the OXTR, which
protects against infarct size in a stroke model [158]. In vivo,
microglia play an important role after stroke [165]. In vitro,
microglia express OXTRs and oxytocin pretreatment attenu-
ated the reactivity of these cells to an inflammatory
stimulus (LPS) [158]. Speculatively, these data implicate oxy-
tocin regulation of microglia in the acceleration of stroke
recovery with social support.
The anti-inflammatory effects of oxytocin/OXTR acti-
vation on microglia appear to be conserved across species
and in development. For example, chronic hydrocortisone
treatment in zebrafish larvae induces microglia activation
(including increases in IL-1β, IL-8 and TNFα expression,
and increased activated amoeboid microglia). Activating the
OXTR with carbetocin treatment blocked this hydrocorti-
sone-induced inflammatory microglia activation [166]. A
similar effect was observed in mice. Compared to wild-type
mice, adult male OXTR knockout mice displayed increased
activation of microglia as assessed by Iba1 immunoreactivity
 in the medial amygdala and the lateral septum, but not the
medial prefrontal cortex [167]. Additionally, these OXTR
knockout mice had decreased levels of the synaptic scaffold-
ing protein, PSD95, measured in whole-brain preparations.
This genetic effect was rescued with pharmacological silen-
cing of microglia with minocycline throughout pre- and
postnatal development. These findings are consistent with a
direct effect of OXTR providing negative feedback regulation
to constrain the impact of developmental microglial acti-
vation on synaptic modulation [167]. A large body of
evidence has suggested alterations to the postsynaptic den-
sity are associated with the development of ASD (reviewed
in [168]). These important clues continue to point to the
need to better understand mechanisms of bidirectional regu-
lation of oxytocin and microglia in health and disease.
Both in vivo and in vitro evidence across the lifespan and
across species indicate causal bidirectional regulatory inter-
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action between oxytocin and microglia. This raises the
question of how their interaction in development may be
important for social behaviour—expressed during develop-
ment as well as later in adulthood.
6. Microglia and oxytocin in developmental-
sensitive periods with social behaviour
implications
Oxytocin is an important regulator of social behaviour,
including pair bonding between partners, sociability between
conspecifics, social recognition, aggression, parental care and
caregiver–infant attachment (reviewed in [8]) and emerging
evidence indicates a role for oxytocin in infant attachment
to a caregiver [169–172]. Oxytocin might execute some of
these functions through its interactions with microglia. Devel-
opmental dysregulation of oxytocin can alter behaviour in
later life, and dysregulation has been associated with
depression [173,174], anxiety [175,176] and ASD [177–179].
Similarly, developmental dysregulation of microglia can dis-
rupt typical development leading to alterations in social
behaviour [180–187]. Finally, social experience in different
phases of development can regulate the oxytocin system
(reviewed in [188]) and microglia (reviewed in [189]).
There are several important developmental periods in
which bidirectional regulation between oxytocin and micro-
glia could influence the developmental trajectory of social
behaviour in health and disease. As seen in adults, the dua-
lity of microglia may contribute to both essential
developmental processes and neuropathology when dysregu-
lated. There is evidence for the interaction of oxytocin and
microglia at prenatal, early postnatal and juvenile stages of
development. In the next sections, we will discuss this sensi-
tive period evidence. Intriguingly, many of the observed
relationships between oxytocin and microglia appear to be
sex-specific.
(a) Prenatal development and birth
Prenatal development involves activation of the innate
immune system with exposures to endogenous fetal com-
ponents such as pulmonary-derived surfactant proteins and
phospholipids in fetal amniotic fluid in the typically sterile
amniotic space (figure 2). The innate immune system is also
activated by exposure to bacteria and viruses. Placental
macrophages are the initial effector cells of this innate 7
immune response, which detect these signals by immune
royalsocietypublishing.org/journal/rstb
mediators, such as toll-like receptor 4 (TLR4) [190]
(figure 2). The activation of TLR4 has been shown to down-
regulate 15-hydroxyprostaglandin dehydrogenase (PGDH)
in fetal membranes and amniotic fluid of mice, though the
mechanism through which this occurs is still unclear [191].
PGDH serves to catalyse the oxidation of prostaglandins
thereby rendering them biologically inactive [192]. A key
prostaglandin during pregnancy and parturition is prosta-
glandin E2 (PGE2), which also plays a role in the
inflammatory response [193]. In addition to the downregula-
tion of PGDH, TLR4 activation upregulates the production of
pro-inflammatory cytokines, such as IL-6, IL-1 [194–196] and
Phil. Trans. R. Soc. B 377: 20210059
TNFα [197–199] (figure 2). Though demonstrated to play
other roles during parturition, oxytocin has been shown to
exhibit anti-inflammatory properties [200] which may serve
to protect the fetus from the increasing inflammatory
environment during typical in utero development.
In utero exposure to infection can have long-term neuro-
developmental [201–203] and behavioural [204] effects
through vertical transmission by placental macrophages
[205], with potential sex-specific effects on some outcomes
[206]. Animal models of in utero flu virus exposure indicate
causal impacts on oxytocin, microglia and social behaviour.
Seasonal flu virus (H3N2) given to first-time pregnant
BALB/c mice on gestational day 9.5 (right after anterior
neural tube closure and during subdivision of forebrain ves-
icles) leads to sex-specific neurochemical, anatomical and
behavioural changes in adult offspring [207]. In young
adult (P56) males and female mice, in utero flu exposure eli-
cited dose-dependent effects on social behaviour and on
aggression and reduced oxytocin and serotonin levels. High
doses of seasonal flu virus increased the Iba1 immunoreactiv-
ity of microglia in the brainstem of males only and decreased
oxytocin levels in plasma in both males and females. Thus, in
utero infection caused long-term changes in microglia and
oxytocin evident in adulthood.
Prenatal malnutrition can also lead to upregulated neu-
roinflammatory pathways in microglia [208]. Malnutrition
combined with infection induces significant changes to typi-
cal developmental trajectories. In a two-hit rat model of fetal
growth restriction ( prenatal low protein diet and postnatal
IL-1β exposure [166]), neonates exhibited reduced Oxt
mRNA levels and increased hypothalamic Oxtr mRNA
levels, and increased markers of inflammation in the cortex.
In sorted microglial cells, microarray analysis indicated
robust upregulation of inflammatory genes. The addition of
an OXTR agonist, carbetocin, alleviated the observed inflam-
matory response by reducing the expression of pro-
inflammatory cytokines, but had no effect on the expression
of anti-inflammatory cytokines. The inflammatory responses
resulting from the two-hit model were associated with later
atypical neurodevelopment. By P10 there were apparent
reductions in myelination after the two-hit insult. On P28,
rats were imaged with functional ultrasound. The two-hit
model impaired interhemispheric and intra-hemispheric con-
nectivity, which was prevented with neonatal carbetocin
treatment. Behaviourally, P28 animals with the two-hit
insult had increased anxiety-like behaviours in the open
field test, which were prevented with neonatal carbetocin
treatment. Therefore, rats exposed prenatally to a low protein
diet, combined with an inflammatory insult in the early
 postnatal period, had increased microglia activation and
long-term anatomical and behavioural changes that were
ameliorated with early postnatal OXTR activation at the
time of the second insult [166].
More recent evidence indicates that environmental insults
such as cold stress during prenatal development can lead to
microglia activation in the hypothalamus, impairment in oxy-
tocin production in development, and reduced sociability in
adulthood [209]. Cold stress (exposure to 4°C for 30 min)
was induced in pregnant female mice beginning on embryo-
nic day E11.5 and repeated every day through E15.5. Cold
stress reduced the number of oxytocin-positive cells in the
male PVN to about 25% of unstressed controls without sig-
nificantly reducing the numbers of oxytocin-positive cells in
females. At E15.5, four cluster types of microglia were ident-
ified from the hypothalamus based on their unique
expression profiles. Of the four clusters, ‘cluster 2’ cells
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were predominantly located next to the third ventricle and
also included a smaller pool of cells located in the supraoptic
nucleus on the pial side, and also surrounding the anterior
commissure. Cluster 2 cells were observed to contact a
neural stem cell pool, raising the intriguing possibility that
microglia might be involved in regulating the types of cells
emerging from the neural stem cell pool. Indeed, in vitro
experiments demonstrated that a marker enriched only in
males after cold stress, CCL4, increased the proliferation of
progenitor cells. The impairment in oxytocin number in
males was rescued with a maternal diet including PLX622,
a CSF1-receptor inhibitor that depletes greater than 99% of
microglia, indicating that the changes in microglia after cold
stress could be the mechanism responsible for reducing oxy-
tocin production in males, although the drug also affects
other cell types in the periphery [210]. The drug alone did
not alter oxytocin cell number in males or females. In
addition to cold stress in utero impairing oxytocin production
in males, CCL3/4 in utero also impaired oxytocin cell
number, and this effect was present in both males and
females. CCL3 and CCL4 share a receptor called CCR5. A
selective CCR5 antagonist, DAPTA, given to dams during
the cold stress was able to prevent the oxytocin impairment
in males. With these sex-specific changes in anatomy after
cold stress, this begs the question of the potential behavioural
impacts after cold stress. Cold stress impaired sociability in
males, which was rescued by a PLX5622 diet. Cold stress
also impaired sociability in females, which was not rescued
with diet, suggesting a different aetiology. A working
model of the combined data suggests that cold stress causes
microglia cluster type-2 cells to release more cytokines (like
CCL4) in males only. CCL4 binds to CCR5 on neural stem
cells in the progenitor pool along the third ventricle. This
leads to underproduction of oxytocin neurons and oligoden-
drocytes—but only in males. In adulthood, in utero cold
stressed males have an impairment in sociability, but not if
the entire pool of microglia are inhibited during the same
time as the cold stress in utero. Cold stress does impair socia-
bility in females, but not through microglia reducing oxytocin
production or oligodendrocytes. A separate, as yet unknown,
mechanism induces sociability impairment in females after
in utero cold stress.
Adverse neurodevelopmental outcomes after in utero
exposures to pharmacotherapeutics or their metabolites are
possible when drugs are needed for maternal health. A pro-
minent example of this is Depakote (valproic acid)
treatment for epilepsy, migraine or bipolar disorder. Children 8
born to mothers treated with valproic acid have an increased
royalsocietypublishing.org/journal/rstb
risk of neurodevelopmental disorders including ASD [211].
Valproic acid treatment in animal models indicates a causal
influence on brain development [5]. Prenatal treatment with
valproic acid causes social impairment in adulthood and
leads to upregulation of pro-inflammatory markers like
TNFα, IL-1β and IL-6 in juvenile mice and an increase in
Iba-1 expressing microglia [212]. Intranasal administration
of oxytocin at postnatal day 30 in valproic-acid-exposed
mice alleviated all transcriptional markers of pro-inflam-
mation and alleviated the number of Iba-1 microglia.
Repetitive behaviours and social interaction were also
rescued with acute oxytocin treatment.
Phil. Trans. R. Soc. B 377: 20210059
In utero exposures that engage the innate immune system
and/or impair the oxytocin system may set up further vulner-
abilities that exacerbate the challenges of birth. Birth represents
a unique transition in the life of the organism from an aquatic
to terrestrial environment [213]. Numerous challenges are pre-
sent among regulatory systems. Hypoxia, contractions,
passage through the birth canal (or not if the birth mode is
Cesarean delivery) and inoculation with commensal bacteria
from the maternal microbiome are a few of the major events
at birth. The inflammatory insult of birth, if sustained, can
cause dysregulation of the neonatal immune system and con-
tribute to children’s mental health disorders. Prenatal
exposure to valproic acid or OXTR antagonist can impair the
birth transition. Prenatal valproic acid treatment impairs the
neuroprotective transient switch of the neurotransmitter
GABA from excitatory to inhibitory at birth [214]. Similar
effects were observed when an OXTR antagonist [214] was
given to the pregnant mother via the drinking water on the
day before delivery. In combination with maternal immune
activation, neonatal hypoxia ischemia, as a result of parturi-
tion, increases the NF-κB signalling pathway, resulting in
prolonged neuroinflammation. The increased neuroinflamma-
tory environment results in decreased synaptic density in the
hippocampus of neonatal mice [215]. Oxytocin may serve as
a protective anti-inflammatory during birth to restrain micro-
glia [14].
(b) Postnatal attachment and socialization
Infants form social attachments to their caregivers, through
an experience-dependent process. An early precursor to
attachment must involve learning about sensory stimuli
(what is social versus what is not). Once that is established,
the infant must learn how to use a caregiver to regulate its
own distress. Learned attachment styles play an important
role in the infant’s social and emotional capability later in
life [216,217]. Studies in rodents have demonstrated secure
social attachments act as a social buffer and alleviate some
of the stress response in part by modulating the PVN and
corticosterone output of the hypothalamic–pituitary–adrenal
axis. [218–220].The activity of the PVN in neonatal rat pups
was shown to be directly related to the presence of the
mother when presented with a stressful stimulus [220]. In
mouse models of early life stress using brief daily maternal
separation (DBS) from postnatal day 1 to 21, plasma corticos-
terone levels were higher in DBS pups than controls, a pattern
that continued into postnatal day 14 [221]. Another study
using the same early life stress paradigm demonstrated an
upregulation of corticosterone and a downregulation of
 LPS-binding protein, a protein that mediates phagocytosis in
innate immune cells, including microglia. This downregula-
tion resulted in decreased pruning of CA1 pyramidal
neurons in the hippocampus of postnatal day 30 male mice.
Behavioural deficits were also noted in these mice in adult-
hood, specifically increased anxiety-like behaviours [222]. In
male mice, 3 hour daily separations from postnatal day 2 to
14 increased levels of neuroinflammatory markers in the hip-
pocampus and depression-like behaviour in adulthood.
Treatment with oxytocin (delivered intracerebroventricularly)
just prior to testing in adulthood rescued the depression-like
behaviour and normalized the levels of neuroinflammatory
markers [188].
Peripheral stimuli that cause pain and local inflammation
in the neonatal period can have long-term effects on oxytocin
and microglia in the brain as well as later emerging social be-
haviour [223]. For example, formalin injected into the two
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hind paws of neonatal Wistar rats results in a biphasic
response. The first response occurs within 5 min and is related
to the somatosensory activation of pain receptors. The second
occurs over a period of more than 20 min and is a result of an
inflammatory reaction in the peripheral tissue and activity
changes in the spinal cord where sensory input enters the
spinal cord. When rat pups were evaluated on postnatal day
6 after 3 days of treatment from day 3–5, there was elevated
mRNA in blood for TNFα and IL-1β in males and females,
with no changes in IL-6. In the brain, mRNA for TNFα
increased for males but not females. In the brain, there were
no neonatal changes in IL-1β or IL-6. In addition to increased
TNFα in the brain of males, there was also elevated Iba-1
immunoreactivity (activated microglia) in cortex and
increased markers of cell death in the cortex and CA2 of hip-
pocampus. Many of these outcomes seemed to be prevented
with NSAID treatment (indomethacin). Later outcomes result-
ing from formalin injection on postnatal days 3–5 measured at
postnatal day 21, showed more male-specific effects. In blood
and brain tissue, mRNA for TNFα and IL-1β was elevated in
males but not females. Structural changes were also observed:
formalin injection reduced axonal diameter in cortical layers
IV/V and reduced neurogenesis in the dentate gyrus. By wes-
tern blot, oxytocin levels (likely measured by neurophysin I,
instead of actual oxytocin) were decreased in male cortex.
Indomethacin co-applied with formalin blocked these effects
in the cortex. OXTR levels in hippocampus CA1 and CA2
were reduced by formalin in males and females, although
the effect was larger in males. These molecular and anatom-
ical changes after neonatal peripheral inflammatory injury
were associated with changes in social behaviour evaluated
during the juvenile period. In particular, juveniles treated neo-
natally with formalin injections showed impaired sociability,
poor social novelty preference and impaired social recognition
memory. Males were more impaired than females. To sum-
marize, males had activated microglia and decreased
markers of oxytocin peptide in the cortex at P21 after 3 days
of neonatal treatment with formalin injections. Sociability
and social novelty preference were impaired, as was social rec-
ognition behaviour. Thus, a neonatal inflammatory event led
to changes in oxytocin, OXTR and social behaviour that was
apparent as the developing rat entered into another develop-
mental-sensitive period, creating the potential for ongoing
developmental impairment as the juvenile socialization
period uses social experiences to further refine the develop-
ment of the brain.
The juvenile brain undergoes experience-dependent 9
changes. Social experience during this period is an important
royalsocietypublishing.org/journal/rstb
contributor to adult social preferences and the quality of
adult behaviour. Rats engage in robust rough and tumble
play during this socialization-sensitive period. Social
interaction during the post-weaning period of juvenile devel-
opment is necessary for the full species-typical expression of
social behaviour. In rats, social isolation (versus group hous-
ing) for two weeks after weaning from the home cage
reduced vasopressin immunoreactive cell number in the
PVN in males and oxytocin immunoreactive cell number in
the PVN in females [224]. Isolation during this time period
also impacted social recognition behaviour. The lack of
social interaction during this period impacts immune
Phil. Trans. R. Soc. B 377: 20210059
system function in the brain and periphery [225]. Specifically,
social isolation in the adolescent period increases markers of
neuroinflammation and impairs the oxytocin system, with
potential sex-specific effects. Normal sex differences of motiv-
ated social behaviour during adolescence are, in part, due
to the phagocytosis by microglia of dopamine 1 receptors
in the nucleus accumbens [182]. This microglial process
causes the developmentally typical reduction in social play
behaviour in juvenile male rats as they reach the end of
adolescence.
Several gene variants have been associated with ASD,
including phosphatase and tensin homologue (PTEN). In a
mouse model of mutant PTEN, behavioural deficits were
observed in juveniles, including decreased sociability and a
preference for familiar social stimuli instead of the typical
preference for novel social stimuli. These juveniles also dis-
played an increase in repetitive behaviour [226].
Dysregulated PTEN resulted in a significant increase of phos-
phorylated protein kinase B (Akt) which activates NF-κB. As
such, microglial cells become activated and release pro-
inflammatory cytokines. Oxytocin components (i.e. neuro-
physin I immunoreactivity) and oxytocin mRNA levels
were increased, as were several markers of active microglia.
Interestingly, another study using an autism mouse model
found that administration of oxytocin alleviates oxidative
stress, inflammation and repetitive behaviours [212]. It is
possible the differences observed in these two studies is
due to the use of different autism models. It is also possible
that the endogenous increase in oxytocin observed by Sarn
et al. [226] serves to combat overactivation of microglia
induced by the depletion of PTEN [226], or perhaps, alterna-
tively, reflects an inability to use oxytocin or cleave it
appropriately.
7. Conclusion and future directions
The release of immune mediators by microglia is necessary
for prenatal and postnatal development. However, the over-
activation of microglia by way of pathogen exposure during
development can result in a disturbance of proper synaptic
development, circuit formation and social behaviours. Unco-
vering the mechanisms that contribute to microglia–neuron
crosstalk are essential for a refined understanding of the mol-
ecular regulation of social behaviours and potential
therapeutic targets to alleviate behavioural dysfunction.
However, more research is necessary to validate a causal
relationship between inflammatory activation in utero and
in different postnatal-sensitive periods for altered
 behavioural outcomes later in life. Moreover, the specific con- as we attempt to understand human health and the aetiology 10
tribution of oxytocin and regulation of OXTR in microglial of disease where much of the data in humans is correlational,

cells after immune challenge as it relates to social behaviour
needs to be assessed. Emerging research points to a develop-
mental contribution of commensal organisms (the
microbiome) and oxytocin–microglia interactions
[187,227,228]. Future studies using a combination of primary
cell culture, in vivo molecular manipulation and behavioural
assays could present a more comprehensive determination
of the interplay between oxytocin, microglia and social beha-
viours. Specific attention should be paid to the molecular
phenotype of microglial cells, including tissue location and
local milieu and immune status. Attention must also be
paid to the timing of inflammatory insult or experience as
royalsocietypublishing.org/journal/rstb
and potentially misleading for understanding cause and
effect mechanisms.
While human evidence is often correlational, this does not
mean that causal modelling in animals alone is sufficient. As
a case in point, there is ample causal evidence from pre-clini-
cal animal models that oxytocin modulates social behaviour.
This has led to enthusiasm for oxytocin as a therapeutic for
social behaviour challenges. However, a recent large clinical
trial suggesting no significant benefit to social behaviour of
intranasal oxytocin for children with ASD [230]. This indi-
cates that current methods of off-label use of intranasal
oxytocin is not supported for ASD, but further clarifying

Phil. Trans. R. Soc. B 377: 20210059

it relates to sensitive periods in social behaviour develop- information is needed, such as age, state-dependent oxytocin
ment. Sex differences must continue to be considered. system reactivity, social experience/context, genetic variation
The developmental timing of atypical exposures will and perhaps indicators of innate immune function. Perhaps
Downloaded from https://royalsocietypublishing.org/ on 26 October 2022

likely play a very important role in the impact on develop-
mental outcomes. A recent study in mice [229]
demonstrates the dissociability of outcomes to the same
insult through different mechanisms in different systems.
Pregnant mice treated with an inflammatory agent ( poly I:
C) give birth to offspring who have increased inflammatory
susceptibility and altered neurodevelopmental outcomes,
modelling a repeated observation of co-occurring immuno-
logical and behavioural features in a subset of individuals
diagnosed with ASD. Through careful study design to tease
apart the timing of the effects using cross-fostering, it
became clear that the neurodevelopmental and behavioural
outcomes were attributable to in utero inflammation, while
the immune sensitization was caused by the postnatal micro-
biome status of the rearing dam, which was caused by
maternal inflammation during pregnancy. Thus, a prenatal
immune challenge had more direct impacts on offspring
brain development and delayed indirect effects on the off-
spring immune system through the postnatally transmitted
microbiome. Findings like this will be especially important
oxytocin is not the appropriate intervention for all causes of
ASD, but as we improve our understanding of the causes
of ASD, we may find an appropriate sub-type where oxytocin
may help. Understanding oxytocin and microglia interactions
might lead to improved understanding of the development of
social behaviour, with opportunities for biological interven-
tion when needed if changing the social environment is
insufficient.
Data accessibility. This article has no additional data.
Authors’ contributions. A.G.: visualization, writing—original draft and
writing—review and editing; E.A.D.H.: writing—original draft and
writing—review and editing.
All authors gave final approval for publication and agreed to be
held accountable for the work performed therein.
Conflict of interest declaration. We declare we have no competing interests.
Funding. The authors are supported by NIH MH114994 (to E.A.D.H.)
and the Good Nature Institute (to E.A.D.H.).
Acknowledgements. The authors would like to thank lab members for
helpful discussions on initial drafts of this work.

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