Antidepressant-Like Effects of Medial Prefrontal

Cortex Deep Brain Stimulation in Rats

Clement Hamani, Mustansir Diwan, Carlos E. Macedo, Marcus L. Brandão, Jason Shumake,
Francisco Gonzalez-Lima, Roger Raymond, Andres M. Lozano, Paul J. Fletcher, and José N. Nobrega
Background: Subcallosal cingulate gyrus (SCG) deep brain stimulation (DBS) is being investigated as a treatment for major depression. We
report on the effects of ventromedial prefrontal cortex (vmPFC) DBS in rats, focusing on possible mechanisms involved in an antidepressant-
like response in the forced swim test (FST).

Methods: The outcome of vmPFC stimulation alone or combined with different types of lesions, including serotonin (5-HT) or nore-
pineprhine (NE) depletion, was characterized in the FST. We also explored the effects of DBS on novelty-suppressed feeding, learned
helplessness, and sucrose consumption in animals predisposed to helplessness.

Results: Stimulation at parameters approximating those used in clinical practice induced a significant antidepressant-like response in the
FST. Ventromedial PFC lesions or local muscimol injections did not lead to a similar outcome. However, animals treated with vmPFC ibotenic
acid lesions still responded to DBS, suggesting that the modulation of fiber near the electrodes could play a role in the antidepressant-like
effects of stimulation. Also important was the integrity of the serotonergic system, as the effects of DBS in the FST were completely abolished
in animals bearing 5-HT, but not NE, depleting lesions. In addition, vmPFC stimulation induced a sustained increase in hippocampal 5-HT
levels. Preliminary work with other models showed that DBS was also able to influence specific aspects of depressive-like states in rodents,
including anxiety and anhedonia, but not helplessness.

Conclusions: Our study suggests that vmPFC DBS in rats may be useful to investigate mechanisms involved in the antidepressant effects of
SCG DBS.

Key Words: Brodmann area 25, cingulate gyrus, deep brain stimu- (13–15). Potential antidepressant effects of IL or vmPFC electrical
lation, depression, infralimbic cortex, medial prefrontal cortex stimulation in animals have not been investigated in detail.
Here, we report on the antidepressant-like effects of vmPFC
DBS in the forced swim test (FST), a well-established tool that

D
eep brain stimulation (DBS) is currently being investi-
gated as a potential therapy for treatment-refractory has been successfully used to investigate antidepressant ac-
depression. One of the most promising DBS targets for tivity (16 –24). In addition, we provide preliminary data in
this condition is the subcallosal cingulate gyrus (SCG), including other animal models, suggesting that DBS may influence other
Brodmann area (BA) 25. Overall, 60% of patients treated with aspects of depressive-type states in rodents, including anxiety
stimulation responded to the procedure in the first reported and anhedonia.
clinical series (1,2). Despite these encouraging results, the basic
neural and chemical mechanisms underlying DBS are still un- Methods and Materials
known. One approach to address these issues is to investigate All protocols were approved by the Animal Care committee of
the effects of stimulating homologous regions in experimental the Centre for Addiction and Mental Health.
animals.
The anatomy of the prefrontal cortex varies considerably
Surgical Procedures
across species and the correspondence between different struc-
Male Sprague-Dawley rats (250 to 300 g) were anesthetized
tures is somewhat controversial (3,4). Based on anatomical
with ketamine/xylazine (100/7.5 mg/kg intraperitoneal [IP]) and
connections and cytoarchitectural features, however, the ventral
had electrodes implanted in the vmPFC at the following stereo-
aspect of the medial prefrontal cortex (vmPFC), particularly the
taxic coordinates: anteroposterior (AP) ⫹ 3.0, lateral (L) ⫾ .4, and
infralimbic cortex (IL), is the region most commonly suggested as
depth (D) 5.6 mm (25). Monopolar stainless steel electrodes (125
the anatomical correlate of BA 25 (5,6). In rodents, the IL has
␮m in diameter and .75 mm of exposed surface) were used as
mainly been implied in mechanisms of stress (7–9), autonomic
cathodes. Similar electrodes implanted in the somatosensory
responses (10 –12), and the extinction of conditioned responses
cortex were used as anodes (AP .5, L ⫾ 1.5, D 1.0 mm) (25).
Control animals had holes drilled into the skull but were not
From the Neuroimaging Research Section (CH, MD, RR, JNN) and Biopsy-
implanted with electrodes. Animals receiving striatal stimulation
chology Section (PJF), Centre for Addiction and Mental Health; and
had cathodes implanted in the caudate-putamen (AP .5, L ⫾ 3.5,
Division of Neurosurgery (CH, AML), Toronto Western Hospital, Toronto,
D 4.5 mm) and anodes in the somatosensory cortex (AP ⫺2.5,
Ontario, Canada; Laboratório de Psicobiologia (CEM, MLB), Faculdade de
Filosofia, Ciências e Letras, Universidade de São Paulo, Ribeirão Preto,
L ⫾ 1.5, D 1.0 mm) (25).
Brazil; and Department of Psychology (JS, FG-L), University of Texas at Unless otherwise specified, behavioral experiments were
Austin, Austin, Texas. conducted 7 days after any surgical procedure.
Address correspondence to Clement Hamani, M.D., Ph.D., Neuroimaging
Research Section, Centre for Addiction and Mental Health, 250 Col- Forced Swim Test and Electrical Stimulation
lege Street, Toronto, Ontario M5T 1R8, Canada; E-mail: clement. On the first day of testing, rats were individually placed for 15
hamani@uhn.on.ca. min in a cylinder (25 cm diameter, 60 cm tall) filled with 25°C ⫾
Received Mar 26, 2009; revised Aug 13, 2009; accepted Aug 14, 2009. 1°C water to a depth of 40 cm. Thereafter, DBS-treated animals

0006-3223/10/$36.00 BIOL PSYCHIATRY 2010;67:117–124

doi:10.1016/j.biopsych.2009.08.025 © 2010 Society of Biological Psychiatry
118 BIOL PSYCHIATRY 2010;67:117–124
received continuous stimulation for 4 hours. On the following
day, rats in this group underwent 2 hours of stimulation followed
by a 5-min swimming session. This was divided into 5-sec
segments and the predominant behavior of the animals (immo-
bility, swimming, or climbing) in each segment was blindly
scored (maximum of 60; 1 point per segment) (18,19). In the FST,
low immobility scores are associated with an antidepressant-like
effect.
Stimulation was conducted with a handheld device (St. Jude
Medical model 3510, Plano, Texas) at 100 ␮A, 130 Hz, and 90
␮sec. These settings were chosen, as they would generate a
charge density similar to that observed in patients undergoing
DBS (26). The time frame of stimulation was selected based on
previous literature showing that one to two doses of antidepres-
sants after swimming on day 1 and one dose before swimming
on day 2 reduce immobility (22). In this context, we used twice
as much stimulation on day 1 (4 hours) than on day 2 (2 hours).
Imipramine (15 mg/kg IP, Sigma-Aldrich, Oakville, ON, Can-
ada) or saline was injected 1 hour and 5 hours after swimming on
day 1 and 2 hours before swimming on day 2, as previously
described (22).
Open Field Test
Two days after the FST, animals received stimulation for 4
hours on day 1 and 2 hours on day 2. Thereafter, locomotor
activity was assessed for 30 min in a square .49 m2 open field
(Med Associates, St. Albans, Vermont) with infrared photo beams
placed along the walls of the equipment. Crossing of the beams
provided counts of motor activity.
Radiofrequency Lesions, Ibotenic Acid, and Muscimol
Injections
For radiofrequency (RF) lesions, probes with .125 mm in
diameter (.75 mm of exposed tip) were stereotaxically implanted
in the vmPFC and a radiofrequency generator (Radionics, Burl-
ington, Massachusetts) was used to deliver 20 mA for 15 sec.
Control animals had probes inserted into the target but were not
lesioned.
For ibotenic acid (IBO) lesions, 2.5 ␮g of ibotenic acid (Sigma
Aldrich) was dissolved in .5 ␮L of .1% ascorbic acid and
stereotaxically injected into the vmPFC over 48 sec with an
infusion pump. The needle was left in place for 4 additional
minutes, after which the procedure was repeated in the contralat-
eral side. Control animals were injected with .1% ascorbic acid.
Animals in the IBO ⫹ DBS group had IBO lesions and vmPFC
electrodes implanted during the same surgical procedure.
Animals receiving muscimol were bilaterally implanted with
guide cannulae 1 mm above the target. Three .5-␮L injections of
either 50 ng of muscimol or saline (control animals) were
administered over 2 min through a needle protruding 1 mm from
the tip of the guide cannula. This drug concentration was chosen
based on previous studies in which vmPFC muscimol injections
induced a positive response in animals undergoing controllable
stress and influenced serotonin release by raphe neurons (9).
Animals received muscimol or saline immediately following the
first swimming session, 5 hours later, and 1 hour before the
second swimming session.
5,7-Dihydroxytryptamine Injections and Serotonin Depletion
Selective serotonin (5-HT) depletion was achieved through
the injection of 5,7-dihydroxytryptamine (5,7-DHT) into the
dorsal and median raphe nuclei. A 30-gauge needle was first
lowered into the median raphe (AP ⫺7.8, L ⫾ 0, D 8.8 mm) and
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C. Hamani et al.
.2 ␮L of a solution containing 4 ␮g of 5,7-DHT creatinine sulfite
(Sigma Aldrich) in .1% ascorbic acid was injected over 4 min.
After an additional 4 min, the needle was raised by 2.2 mm and
the process repeated in the dorsal raphe nucleus (27). Control
animals received .1% ascorbic acid. During the same surgical
procedure, animals were either implanted with vmPFC elec-
trodes or had holes drilled into their skull, as described above.
Behavioral experiments using 5-HT depleted animals were con-
ducted 14 days after surgery for the animals to fully recover from
the 5,7-DHT injections.
N-(2-Chloroethyl)-N-Ethyl-2-Bromobenzylamine Injections
and Norepinephrine Depletion
Animals had vmPFC electrodes implanted or holes drilled
into the skull (control animals). On the following day, they
received N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) (50
mg/kg IP) for selective norepinephrine (NE) lesioning or saline.
Microdialysis
In a single procedure, electrodes were bilaterally inserted into
the vmPFC and a microdialysis cannula implanted in the right
dorsal hippocampus (AP ⫺3.7, middle lateral [ML] 2.4, D ⫺5.1
mm) (27). Five days later, a microdialysis probe (CMA/12-2 mm
membrane, CMA Microdialysis, Solna, Sweden) was inserted into
the target and perfused with Ringer’s solution at a constant flow
rate of 1.5 ␮L/min (microinjection pump, BAS, West Lafayette,
Indiana). Following an equilibration period (3 hours), dialysate
samples were collected every 30 min for 2 hours in vials
containing 5 ␮L of perchloric acid solution (.05 N). Baseline
levels were defined as an average of these four samples. Animals
then received stimulation for 1 hour, while sham-treated control
animals had electrodes implanted but were not stimulated.
Dialysate samples continued to be collected for another 180 min.
Details on the monoamine assay and sample analysis have been
previously described (28).
Novelty-Suppressed Feeding (NSF)
Animals were initially habituated to a food reward (chocolate
pellets), which was not provided on the day before the experi-
ments. Ventromedial PFC DBS was carried out for 4 hours on day
1 and 2 hours on day 2. Immediately after stimulation was
discontinued on day 2, animals were placed in a novel Plexiglas
cage (50 ⫻ 22 ⫻ 22 cm) with white walls, which contained a blue
platform with food reward on top. Latency to consume the
reward was measured (sniffing or simply touching the food was
not scored).
Learned Helplessness
Animals were initially placed in sound-attenuated operant
boxes (Med Associates), where they received .8 mA inescapable
shocks. Both the duration (1.5 to 60 sec) and the interval
between shocks (1 to 30 sec) were programmed to result in a
total shock exposure of 25 min per animal. On the following day,
rats were given 15 trials of escapable footshocks (.8 mA, fixed
intertrial interval of 24 sec). These were accompanied by a bright
light, placed directly above a lever; the footshocks could be
terminated by a single lever press. Deep brain stimulation was
applied for 4 hours after inescapable shocks on day 1 and for 2
hours before escapable shocks on day 2.
Sucrose Consumption in Animals Predisposed to Helplessness
Rats predisposed to helplessness (bred at the University of
Texas at Austin) were used in our study because of their lower
sensitivity to sucrose as a reward compared with normal animals
C. Hamani et al. BIOL PSYCHIATRY 2010;67:117–124 119

(29). After 2 hours of stimulation, baseline assessments were
conducted in an operant chamber (Med Associates) with animals
being exposed for 30 min to identical bottles containing a 5%
sucrose solution or water. A light beam passing in front of the tips
of the bottles registered the time spent drinking from each bottle.
Two days later, animals underwent inescapable footshocks (60
10-sec trials of .6 mA separated by 10 to 110 sec), followed the
next day by a session of escapable footshocks (30 trials of up to
30 sec of .6 mA separated by 10 to 50 sec). Deep brain
stimulation was applied for 2 hours before each shock session.
On the following day, a new assessment of sucrose consumption
was carried out in the same operant chambers after 2 hours of
stimulation.
Histology
To assess electrode (Figure 1A) and cannula placement, as
well as location and size of lesions, brains were stained with
cresyl violet. In 5,7-DHT and DSP-4 experiments, the extent and
specificity of 5-HT and NE depletion were evaluated with high
performance liquid chromatography (HPLC) in brain samples
extracted from the frontal cortex, hippocampus, striatum, and
hypothalamus (30) (Figure S1 in Supplement 1).
Statistical Analyses
One- or two-way analyses of variance (ANOVAs) were used
to compare data when three or more independent groups were
considered. For experiments using only two samples, we used
independent or paired Student t test. Statistical significance was
set at p ⫽ .05.
Results
Outcome of vmPFC DBS in the FST
Animals treated with vmPFC DBS (n ⫽ 13) showed a 45%
decrease in immobility scores compared with control animals
(n ⫽ 12; p ⫽ .004; Figure 1B). A similar antidepressant-like
response was not observed after striatal stimulation (n ⫽ 6;
Figure 1C), suggesting that the effects of DBS had some target
specificity. As a positive control, imipramine-treated rats (n ⫽ 8)
had a 55% reduction in immobility scores (p ⫽ .002; Figure 1D),
a magnitude of response similar to that in the DBS group.
To rule out the possibility that reduced immobility in the FST
was due to an overall increase in locomotor activity, animals that
had vmPFC stimulation and control animals were tested in an
open field. No differences were found between groups (p ⫽ .15;
Figure 1E), suggesting that the outcome of DBS in the FST was
not related to a simple locomotor effect.
vmPFC Radiofrequency Lesions, Ibotenic Acid Lesions, and
Muscimol Injections
Because a potential mechanism of DBS is target inactivation
(31), we tested whether the behavioral effects of stimulation

Figure 1. Outcome of ventral medial prefrontal cortex (vmPFC) deep brain stimulation (DBS) in the forced swim test (FST). (A) Schematic representation of
coronal brain sections showing the location of the tip of the electrodes implanted in animals receiving stimulation (light gray circles). Numbers to the right
denote distance from the bregma (reprinted with permission from The Rat Brain in Stereotaxic Coordinates, 3rd ed. by Paxinos and Watson, Copyright Elsevier
[1997]). (B) During behavioral experiments, rats were treated with vmPFC DBS at 130 Hz, 90 ␮sec, and 100 ␮A. For scoring, the predominant behavior
(immobility, swimming, or climbing) during the 5 min of the FST was recorded every 5 sec (maximal score of 60). Animals receiving vmPFC DBS had a
significant reduction in immobility, the hallmark of an antidepressant-like response, compared with control animals (p ⫽ .004). (C) Caudate-putamen (CPu)
DBS did not lead to a similar antidepressant-like outcome, suggesting that the effects of stimulation were target specific. (D) Animals treated with imipramine
(IMI) showed a 55% reduction in immobility compared with saline-treated control animals (p ⫽ .002), a magnitude of response that was similar to the one
observed with DBS. (E) The effects of vmPFC DBS were not likely due to changes in locomotor activity, as performance in these animals was similar to control
animals in an open field. In all figures of the panel, numbers in parenthesis represent the number of animals per group. IL, infralimbic cortex; PL, prelimbic
cortex. *Statistically significant.

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120 BIOL PSYCHIATRY 2010;67:117–124 C. Hamani et al.

Figure 2. Outcome of vmPFC lesions in the FST. Photomicrographs of animals that underwent bilateral radiofrequency (A) or ibotenic acid vmPFC lesions (D).
Note that in both cases, most of the infralimbic cortex, as well as portions of the ventral prelimbic cortex, were severely compromised. While radiofrequency
lesions were characterized by a substantial loss of tissue, ibotenic acid injections often led to significant gliosis. Neither these ablative techniques (B,E) nor the
bilateral injection of muscimol into the vmPFC (C) were associated with a significant antidepressant-like outcome. When animals bearing vmPFC IBO lesions
received stimulation at the same site, however, a significant reduction in immobility scores was observed, compared with IBO-treated nonstimulated rats (p ⬍
.001) or control animals (p ⫽ .02) (D). In all panels, numbers in parenthesis represent the number of animals per group. Arrows in (D) represent the transition
between normal and gliotic tissue after IBO injections. Scale bars in (A) and (D) ⫽ 1 mm. FST, forced swim test; IBO, ibotenic acid; vmPFC, ventral medial
prefrontal cortex. **Statistically significant compared to controls and nonstimulated IBO-treated animals.

could be reproduced by lesioning the vmPFC. Rats received
either bilateral RF lesions to ablate local axons and neuronal cell
bodies (n ⫽ 12) or IBO injections to injure predominantly cell
bodies (n ⫽ 7). In all animals treated with either technique, most
of the IL, and in some cases part of the ventral prelimbic cortex
(vPL), were injured (Figure 2A,D). Neither group had a signifi-
cant antidepressant-like response in the FST (Figure 2B,E),
although RF-lesioned animals showed a 23% nonsignificant
decrease in immobility compared with control animals (p ⫽ .2).
Similar results were observed in animals receiving bilateral
vmPFC muscimol injections. In this group, a 25% nonsignificant
reduction in immobility scores was recorded (p ⫽ .09) (Figure
2C).
We then took advantage of the somewhat selective pattern of
IBO lesions to investigate whether the modulation of potentially
spared vmPFC fibers could contribute to the effects of DBS.
Similar to the response in nonlesioned animals receiving stimu-
lation, IBO-treated rats given vmPFC DBS (n ⫽ 6) had a 47%
decrease in immobility compared with control animals (p ⫽ .02)
(Figure 2E).
Effects of 5-HT and NE Depletion on the Antidepressant-Like
Response of vmPFC DBS in the FST
To begin addressing potential neurochemical substrates for
the efficacy of vmPFC DBS, we studied the effects of stimulation
after 5-HT and NE depletion. We found that the antidepressant-
like response of vmPFC DBS in the FST was completely abol-
ished in rats bearing serotonin- depleting lesions. Reduced
immobility was observed in the vmPFC DBS ⫹ vehicle group
(n ⫽ 6, p ⫽ .02) but not in the vmPFC DBS ⫹ 5,7-DHT treated rats
(n ⫽ 12) (Figure 3A). In contrast, NE depletion did not influence
outcome. No differences were found between animals that had
vmPFC ⫹ saline (n ⫽ 5) or vmPFC ⫹ DSP-4 (n ⫽ 6) (Figure 3B).
In both groups, immobility scores were lower than in saline-
treated control animals (p ⫽ .03, p ⫽ .06, respectively; Figure
3B).
As the antidepressant-like response of vmPFC DBS in the FST
seemed to depend on the integrity of the 5-HT system, we
assessed whether stimulation could influence 5-HT release with
microdialysis. We found that vmPFC DBS for 1 hour (n ⫽ 5)
induced a fourfold increase in 5-HT release (Figure 3C). Return to
baseline levels was only observed 150 min after stimulation was
discontinued, suggesting that 5-HT was still increased when
behavioral assessments were conducted (Figure 3C).
Antidepressant-Like Effects in Other Animal Models
We next carried out preliminary experiments in other animal
models to assess whether stimulation could influence additional
aspects of depressive-type states. We first tested the effects of
DBS on NSF. Though primarily suited to evaluate anxiety, we
chose to use this test, as it is also susceptible to antidepressant
medications (32,33). We found that latency to feed in the
stimulated group was 45% lower than in control animals (p ⫽
.05), suggesting that DBS was associated with an anxiolytic-like
effect (Figure 4A). In contrast, no differences were found in the
latency for an escape response between animals receiving stim-
ulation and control animals in a learned helplessness (LH)
paradigm (p ⫽ .1) (Figure 4B). In animals predisposed to
helplessness, while control animals had a 66% decrease in
sucrose consumption compared with baseline (p ⫽ .01), a 38%

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C. Hamani et al. BIOL PSYCHIATRY 2010;67:117–124 121

Figure 3. Antidepressant-like effects of vmPFC DBS require the integrity of the serotonergic but not the noradrenergic system. (A) Antidepressant-like effects
of vmPFC DBS were not observed in animals given raphe microinjections of 5,7-DHT, a toxin for 5-HT neurons. Decrease in immobility scores only occurred in
animals treated with DBS and raphe vehicle injections (p ⫽ .02 compared with control animals). (B) In contrast, NE depletion did not influence outcome. No
differences were found between animals that had vmPFC ⫹ saline or vmPFC ⫹ DSP-4. (C) vmPFC DBS for 1 hour (horizontal bar) was associated with a fourfold
increase in the levels of hippocampal 5-HT as assessed with microdialysis. A return to baseline levels was only observed 150 min after stimulation was
discontinued. In all figures, numbers in parenthesis represent the number of animals per group. Veh ⫽ ascorbic acid vehicle injections. 5-HT, serotonin;
5,7-DHT, 5,7-dihydroxytryptamine; DBS, deep brain stimulation; DSP-4, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine; NE, norepinephrine; vmPFC, ventral
medial prefrontal cortex. *Statistically significant.

nonsignificant decrease was observed in animals receiving
vmPFC DBS (p ⫽ .2; Figure 4C). This provides preliminary
evidence suggesting that vmPFC DBS may influence hedonic
states in rodents.
Discussion
We found that vmPFC DBS induced a clear antidepressant-
like response in the FST, a well-established paradigm to assess
antidepressant activity in rodents (16 –20). In addition to different
classes of medications, interventions such as electroconvulsive
therapy and vagus nerve stimulation have all shown antidepres-
sant-like effects in this model (16 –21,24,34). While the predictive
validity of the FST is strong, a common criticism to its face
validity refers to the short time frame needed for antidepressant-
like responses to emerge. In this context, it may be interesting to
note that acute responses to DBS have been reported in patients
as soon as SCG electrodes are turned on (e.g., an increase in
energy, a reduction in the sensation of void, among others) (1).
This suggests that the dynamics for an antidepressant response
after DBS and regular medications, which often require weeks
for an adequate response, may not be exactly the same.
In our study, stimulation parameters were chosen to approx-
imate those used in clinical practice. Taking into account elec-
trode diameter and exposed surface, it was estimated that 100 to
300 ␮A would be necessary to deliver a charge density per mm2
of tissue comparable with that in patients with depression
receiving DBS (1). We decided to use the lowest intensity in this
range to minimize current spread and compensate for the smaller
brain volume in rats. We are exploring the effects of different
stimulation parameters in the FST and have preliminary evidence
suggesting that the most effective settings are indeed similar to
those used in clinical practice (e.g., current intensity of 100 to 300
␮A, 90 ␮sec, 130 Hz). Also to be addressed in further detail in our
study is target specificity and the potential current spread to
adjacent structures. Bearing in mind the configuration of our
electrodes and the dimensions of the rat vmPFC, it is likely that
both IL and vPL were somewhat influenced by stimulation in
most animals. In this context, while DBS in distant regions (e.g.,
striatum) did not induce an antidepressant-like effect, we could
not assess potential outcome differences related to stimulation of
different subregions within the vmPFC. We chose to cover a
larger area of the vmPFC based on clinical studies showing that
SCG electrodes in patients with depression were not only
implanted in BA 25 but also in BA 32 and subgenual BA 24 (35).
In addition, the vmPFC in rodents is thought to represent a
functional unit distinct from the dorsal medial prefrontal cortex
(36,37) (though differences between prelimbic cortex and IL
stimulation have been reported in experimental models of fear
extinction [38]). Further studies using electrodes with a smaller
exposed surface will be needed to explore whether outcome in
the FST may change because of target variability within the
vmPFC.

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122 BIOL PSYCHIATRY 2010;67:117–124
Figure 4. Outcome of vmPFC DBS in other animal models relevant to de-
pression. (A) Anxiolytic-like effect of vmPFC DBS in the novelty-suppressed
feeding test. Note the significantly lower latency to eat a reward in animals
receiving stimulation compared with control animals (p ⫽ .05). (B) Effects of
stimulation in a learned helplessness paradigm. No differences were found
between animals receiving DBS and control animals in the latency to escape
the shocks. Naïve rats that were not subjected to inescapable shocks had a
significantly lower escape latency compared with both control animals and
the DBS group (p ⬍ .001). (C) Sucrose consumption in congenitally helpless
animals before and after footshock stress. At baseline, no differences were
found across groups. After stress, however, while control animals had a 66%
reduction in sucrose consumption compared with baseline (p ⫽ .01), a
nonsignificant 38% decrease was observed in the DBS group (38%; p ⫽ .2). In
all figures, numbers in parenthesis represent the number of animals per
group. DBS, deep brain stimulation; vmPFC, ventral medial prefrontal cor-
tex. *Statistically significant.
Radiofrequency lesions were extensively used in functional
neurosurgery before DBS. Indeed, it was the similarity in out-
come between these two therapies in the field of movement
disorders that led to the initial thinking that the functional
inactivation of stimulated targets could be one of the mecha-
nisms of DBS (39). With this in mind, we explored the effects of
vmPFC RF lesions in the FST. In contrast to most clinical
applications of DBS, vmPFC lesions and stimulation were not
associated with a similar magnitude of behavioral response. In
addition, significant responses in the FST were not observed after
either vmPFC IBO lesions or muscimol injections. That being
said, the 25% nonsignificant decrease in immobility scores after
RF and muscimol suggests that target inactivation may have
contributed to some extent to effects of DBS. Clearly, however,
additional mechanisms were involved, considering that the re-
duction in immobility scores was consistently higher after stim-
ulation (i.e., in the order of 45%). An intriguing observation was
that the outcome after IBO ⫹ DBS was similar to that of DBS
alone. Since one of the most commonly proposed mechanisms of
DBS is to drive local axonal projections (31,40), these results
raise the possibility that the modulation of fibers near the
electrodes could, at least in part, be involved in the antidepres-
sant-like effects of stimulation. As efferents from target regions
degenerate after IBO injections, preserved afferents to the IL/vPL
and en-passant fibers might be implicated. Further work is
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C. Hamani et al.
needed to elucidate the nature of pathways and neural circuitry
involved in the mechanisms of DBS.
Another possibility would be that plastic changes taking place
after IBO (41,42), including the reorganization of fiber pathways
innervating the lesioned site (43), could have contributed to our
results. This seems unlikely, however, considering the relatively
short time frame between lesions and behavioral tests in our
study (i.e., 1 week would probably not be enough for the
formation of new functional circuits within the injured site). In
addition, we recorded a similar magnitude of response in animals
treated with IBO ⫹ DBS and DBS alone. Whether plasticity after
IBO had played a major role, results should have been different
in lesioned and nonlesioned animals (as no plastic changes
would occur in this latter group). As vmPFC fibers were the
common element to be potentially influenced by stimulation in
both groups, we suggest that the modulation of these pathways
would comprise a more likely explanation to our findings.
Also, to be considered is the possible involvement of glial
cells, which are preserved after IBO lesions (41,44). This is of
interest, as glial ablation in the prefrontal cortex has been
associated with depressive-like symptoms in rodents (44). Fur-
ther, a reduction in the number of glial cells has been suggested
as one of the potential mechanisms for a smaller gray matter
volume in the SCG of patients with familial depression (45).
Whether the modulation of glial cells plays a role in the
antidepressant-like effects of stimulation remains to be deter-
mined.
As an initial step in assessing potential neurochemical sub-
strates for the effects of DBS, we chose to investigate cat-
echolaminergic systems for several reasons. Both 5-HT and NE
are involved in antidepressant responses in humans and in
animal models. Antidepressants with serotonergic and noradren-
ergic activity, including selective 5-HT reuptake inhibitors (SSRIs)
and selective NE reuptake inhibitors (SNRIs), decrease immobil-
ity in the FST (18,22,34). The vmPFC has direct projections to the
raphe and locus coeruleus and modulates neuronal activity in
both structures (6,46 –50). Infralimbic cortex stimulation in non-
human primates and rodents increases 5-HT release in various
brain regions (51,52). We found that the effects of vmPFC DBS
were completely abolished after 5-HT but not after NE depletion.
This is similar to what has been reported with antidepressant
medications. While 5-HT depletion per se does not reduce
immobility scores, it abolished the antidepressant-like effects of
SSRIs (53).
To study stimulation-induced 5-HT release, we chose the
hippocampus as an initial target, as this structure has been
extensively involved in the pathophysiology of depressive states
(54 –56). We found that vmPFC stimulation at parameters used in
our behavioral tests induced a significant increase in 5-HT levels.
These results do not necessarily imply that the antidepressant-
like effects of DBS in the FST were due to an increase in 5-HT
release, though this is a possibility. The current findings only
show that the effects of vmPFC DBS depend on the integrity of
the serotonergic system and that hippocampal 5-HT levels were
significantly increased after stimulation.
In the final part of our study, we investigated whether the
effects of vmPFC DBS were restricted to the FST or could be
extended to other animal models. Our preliminary findings
suggest some selectivity of the procedure, with stimulation
leading to an anxiolytic effect in the NSF and influencing sucrose
consumption in animals predisposed to helplessness. In contrast,
no significant effect was observed in a LH paradigm. Of note, we
only tested a limited number of stimulation settings in a single LH
C. Hamani et al.
paradigm. It would be important to characterize the effects of
different stimulation parameters (e.g., current intensity, pulse
width, frequency), in other LH paradigms (e.g., shuttle box
responses), within a different time frame (e.g., DBS being
provided for a few days). Also, to be explored is whether
stimulation may interfere with controllable stress. In rodents,
vmPFC muscimol injections have been shown to disrupt control
over stressful situations and influence 5-HT release by the dorsal
raphe (9).
Aside from anxiety and helplessness, we also explored
whether DBS could influence hedonic states in rats predisposed
to helplessness (29). Animals receiving stimulation before stress
did not have an increase in sucrose consumption. This was not
unexpected, as antidepressants also do not increase responsivity
to sucrose in naïve rats (57,58). In our study, a positive outcome
with DBS was only observed after the animals received a series
of footshocks. While control animals had a significant reduction
in sucrose intake, this was milder and nonsignificant in the
DBS-treated group. In the future, it will be important to corrob-
orate these findings in other models of anhedonia, such as
chronic unpredictable stress (44).
In summary, this initial series of studies indicates that vmPFC
stimulation affects several depressive-like symptoms in rats. In
the FST, DBS induced a clear antidepressant-like response that
was dependent on the integrity of serotonergic systems and
likely involved the modulation of fiber tracks in the vicinity of the
electrodes. Whether similar mechanisms play a role in the
antidepressant mechanisms of DBS in humans has to be ad-
dressed in the future. The current results do suggest, however,
that vmPFC DBS in rats may provide a useful model for investi-
gating mechanisms involved in antidepressant DBS effects.
This work was supported in part by funds from the Ontario
Mental Health Foundation.
We thank J.W. Chambers, S. Guirguis, S.J. Harrison, B.
Raczynska, S. Li, and S. Isabella for the technical help.
CH is a consultant for St. Jude Medical. AML is a consultant
for St. Jude Medical, Medtronic, and Boston Scientific. The other
authors report no biomedical financial interests or potential
conflicts of interest.
Supplementary material cited in this article is available
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