Brain, Behavior, and Immunity 89 (2020) 339–349

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Brain, Behavior, and Immunity

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

Decreased motor impulsivity following chronic lithium treatment in male T

rats is associated with reduced levels of pro-inflammatory cytokines in the
orbitofrontal cortex
Wendy K. Adamsa,b, Dominique L. Levesquea, Paul J. Cockera, Sukhbir Kaura, Tamara S. Bodnarc,
⁎
Allan H. Youngb,1, Catharine A. Winstanleya,b,
a
Department of Psychology, Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, BC, Canada
b
UBC Institute of Mental Health, University of British Columbia, Vancouver, BC, Canada
c
Department of Cellular and Physiological Sciences, University of British Columbia, Vancouver, BC, Canada

A R T I C LE I N FO A B S T R A C T

Keywords: Lithium’s efficacy in reducing both symptom severity in bipolar disorder (BD) and suicide risk across clinical
Lithium populations may reflect its ability to reduce impulsivity. Changes in immune markers are associated with BD and
Five-choice serial reaction time task suicidality yet their exact role in symptom expression remains unknown. Evidence also suggests that lithium may
Impulsivity decrease levels of pro-inflammatory cytokines in the periphery and central nervous system, and that such
Neuroinflammation
changes are related to its therapeutic efficacy. However, issues of cause and effect are hard to infer from clinical
IL-1β
data alone. Here, we investigated the effects of chronic dietary lithium treatment on rats’ performance of the 5-
IL-6
RANTES Choice Serial Reaction Time Task (5CSRTT), a well-validated operant behavioural task measuring aspects of
Orbitofrontal cortex impulsivity, attention and motivation. Male Long-Evans rats received a diet supplemented with 0.3% LiCl
Prefrontal cortex (n = 13), or the equivalent control diet (n = 16), during behavioural testing. Blood and brain tissue samples
Striatum were assayed for a wide range of cytokines once any changes in impulsivity became significant. After 12 weeks,
chronic lithium treatment reduced levels of motor impulsivity, as indexed by premature responses in the
5CSRTT; measures of sustained attention and motivation were unaffected. Plasma levels of IL-1β, IL-10 and
RANTES (CCL-5) were reduced in lithium-treated rats at this time point. IL-1β, IL-6 and RANTES were also
reduced selectively within the orbitofrontal cortex of lithium-treated rats, whereas cytokine levels in the medial
prefrontal cortex and nucleus accumbens were comparable with control subjects. These results are consistent
with the hypothesis that lithium may improve impulse control deficits in clinical populations by minimising the
effects of pro-inflammatory signalling on neuronal activity, particularly within the orbitofrontal cortex.

1. Introduction range. Understanding how lithium acts in the brain to improve BD

Bipolar disorder (BD) is a highly debilitating mood disorder in
which subjects typically experience states of mania and depression in-
terspersed with symptom-free (euthymic) periods (Clark and Goodwin,
2004; Quraishi and Frangou, 2002; Strakowski et al., 2009). Lithium is
one of the most effective treatments for BD (Young and Hammond,
2007; Young, 2009), yet its precise mechanism of action remains elu-
sive (Alda, 2015). In addition to ameliorating the depressive symptoms,
lithium is particularly effective at reducing both the frequency and
severity of manic episodes. However, lithium treatment is associated
with a number of problematic side-effects and has a narrow therapeutic
symptoms could therefore lead to the development of efficacious and
novel treatments with an improved side effect profile (e.g. Barkus et al.,
2018). Animal experiments which assess the effects of lithium on key
symptoms of BD could make a valuable contribution in this regard.
During mania, patients often engage in maladaptive and impulsive
behaviours which can have devastating consequences; high impulsivity
is a significant risk factor for suicide, drug abuse and pathological
gambling, conditions which are all highly co-morbid with BD (Swann,
2009; 2010). One of the most widely recognised forms of impulsivity is
the inability to withhold from making a prepotent motor response, also
known as motor impulsivity, impulsive action, or behavioural

⁎
Corresponding author at: Dr. Catharine Winstanley, Department of Psychology, University of British Columbia, Djavad Mowafaghian Centre for Brain Health,
2215 Wesbrook Mall, Vancouver, BC V6T 1Z3, Canada.
E-mail address: cwinstanley@psych.ubc.ca (C.A. Winstanley).
1
Current address: Centre for Affective Disorders, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, UK.

https://doi.org/10.1016/j.bbi.2020.07.018
Received 22 April 2020; Received in revised form 30 June 2020; Accepted 13 July 2020
Available online 17 July 2020
0889-1591/ © 2020 Published by Elsevier Inc.
W.K. Adams, et al.
disinhibition (Evenden, 1999; Moeller et al., 2001). The five-choice
serial reaction time task (5CSRTT) was designed as a rodent analogue of
the continuous performance test used clinically to assess this form of
impulsivity (Rosvold et al., 1956), and which is sensitive to impulse
control deficits in BD (Strakowski et al., 2010; Strakowski et al., 2009;
Swann et al., 2004; Swann et al., 2003), plus its translational validity
has recently been further confirmed through successful back-translation
into humans (Sanchez-Roige et al., 2014; Voon et al., 2014; Young
et al., 2020). In the 5CSRTT, rats learn to detect and respond to a brief
visual stimulus which can occur in one of five spatial locations (Carli
et al., 1983). Responses made prematurely, before the light has been
presented, provide a reliable index of motor impulsivity, and are
mediated by regions of both medial and orbitofrontal cortex analogous
to those implicated in mediating high impulsivity in BD (Blumberg
et al., 1999; Cerullo et al., 2009; Clark et al., 2009). Acute dosing with
lithium can decrease premature responding in rats (Ohmura et al.,
2012), but the effects of chronic dosing regimens more closely modeling
clinical administration have yet to be reported (Keck et al., 1996).
There are many hypotheses regarding the mechanism underlying
lithium’s therapeutic effects. For example, data from in vitro and in vivo
experiments have demonstrated that lithium can alter neurotransmitter
receptor expression and related signal transduction cascades, ion
transport across the neuronal membrane, mitochondrial function, oxi-
dative stress, and hormonal and circadian regulation (Alda, 2015; Einat
et al., 2003; Lenox and Wang, 2003; Manji and Lenox, 1999; 2000;
Manji et al., 1995). It is increasingly recognised that immune signalling
molecules, such as cytokines and chemokines, can influence neuronal
functioning within the central nervous system, and recent data have
also highlighted a potentially important role for neuroinflammation in
both BD and the therapeutic response to lithium (Ayorech et al., 2015;
Dantzer et al., 2008; Lauterbach, 2016; Maddu and Raghavendra, 2015;
Nassar and Azab, 2014; Raghavendra et al., 2014). Somewhat in par-
allel, evidence is accumulating that implicates inflammatory processes
in other conditions characterised by high impulsivity, such as psy-
chostimulant addiction, alcoholism, suicide, and obesity (Brundin et al.,
2017; Cox et al., 2015; Crews et al., 2011; Lewitus et al., 2016). Fur-
thermore, some studies have observed that peripheral markers of neu-
roinflammation track the expression of impulsivity and related per-
sonality traits in healthy volunteers as well as clinical populations
(Juengst et al., 2014; Sutin et al., 2012). While the long-lasting in-
creases in impulsivity observed in a rat model of experimental trau-
matic brain injury strongly correlate with levels of pro-inflammatory
cytokines (Vonder Haar et al., 2019; Vonder Haar et al., 2016; Vonder
Haar et al., 2017), whether lithium’s ability to reduce impulsivity in
rats is associated with altered inflammatory status is unknown.
We therefore conducted the following experiments to test the hy-
potheses that a) chronic administration of lithium would reduce im-
pulsive responses in rats performing the 5CSRTT, and b) such im-
provements in impulse control would be associated with reductions in
brain and plasma levels of inflammatory cytokines and related signaling
molecules.
2. Materials and methods
2.1. Animals
Subjects were 32 male Long-Evans rats (Charles River Laboratories,
Saint-Constant, QC, Canada) weighing approximately 300 g upon ar-
rival. Animals were pair-housed in a temperature-controlled colony
room under a reverse 12 hr light–dark cycle (~22 °C; lights off at
8:00AM). For the first week, animals received free access to standard
chow and tap water to acclimate to the facility, after which they were
gradually food-restricted over a period of 7 days to the maintenance
amount of 14 g/day required for operant testing. In week three, animals
were placed on their respective custom-formulated lithium or control
diets (detailed in section below), on which they remained for the
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Brain, Behavior, and Immunity 89 (2020) 339–349
duration of the experiment. Behavioural training began two weeks later
(i.e. four weeks after arrival). All experimental procedures were con-
ducted in accordance with the standards of the Canadian Council of
Animal Care and were approved by the Animal Care Committee of the
University of British Columbia.
2.2. Lithium-supplemented diet
Rats were randomly assigned to receive either a custom-formulated
diet containing 0.3% lithium chloride (LiCl) or the equivalent control
diet (n = 16 per group; Teklad Custom Research Diets: TD.09574 0.3%
w/v LiCl Diet or TD.02027 Rodent Control Diet, Harlan Laboratories,
Indianapolis, IN, USA). The concentration of LiCl in the diet (2.91 g/kg)
was determined on the basis of pilot experiments indicating that serum
lithium levels fell within the clinical therapeutic range (0.5–1.5 mmol/
L; Levesque, 2013) in the 24 h following ingestion of the 14 g food
allowance. Both control and lithium-treated rats were provided free
access to 0.9% NaCl solution in addition to tap water to minimise ionic
imbalances resulting from the diuretic properties of lithium (Maletzky
and Shore, 1978), and to reduce renal toxicity associated with chronic
lithium treatment (Smith and Amdisen, 1983). During the initial ex-
posure period, two animals responded adversely to the lithium diet and
were humanely euthanised (group numbers at the start of 5CSRTT
training: n = 16 controls, n = 14 lithium-treated). Body weight was
frequently monitored throughout the experiment; instances of severe
weight loss were treated by boosting daily food allowances with the
control diet (~3–6 g), as appropriate. Animals were fed each day after
behavioural testing, and at approximately the same time on non-testing
days in efforts to maintain steady state levels of circulating lithium.
2.3. 5-Choice serial reaction time task (5CSRTT)
Behavioural testing took place in standard five-hole operant cham-
bers enclosed within ventilated, sound-attenuating cabinets (Med
Associates Inc., Fairfax, VT, USA), as previously described (Adams
et al., 2017). In brief, rats were trained to make a nose-poke response to
a 0.5 sec light stimulus randomly presented in one of five holes. Each
session comprised 100 trials and lasted up to 30 min. A nose-poke at the
food tray initiated a trial, after which a 5 sec inter-trial interval (ITI)
ensued before presentation of the stimulus light. A ‘correct response’
was rewarded with a sweetened food pellet (45 mg Dustless Precision
Pellets®, Bioserv, Frenchtown, NJ, USA). A response made in the ITI
before stimulus presentation (‘premature response’), an incorrect, or a
lack of response (‘omission’), were all punished with a 5 sec time-out,
during which the houselight was illuminated and no further trials could
be initiated. Other variables included repeated, ‘perseverative’ re-
sponding at the nosepoke holes, and response latencies.
Animals were trained for 4–5 sessions/week until they reached the
final training stage (i.e. 0.5 sec stimulus light duration), which required
37 sessions in this cohort (8 weeks of training). One lithium-treated rat
was excluded from analyses as it was unable to learn the task (final
group numbers: n = 16 controls, n = 13 lithium-treated). Chronic li-
thium exposure did not otherwise alter rats’ ability to learn the 5CSRTT,
as both groups completed a similar number of training sessions
(Control: 20.8 ± 1.7; Lithium: 22.8 ± 2.3). Behavioural data from
session 38 onwards were assessed for stability through repeated-mea-
sures analysis of variance (ANOVA) over the most recent 3–5 sessions
with Session as a within-subjects factor. Null effects of Session were
observed by session 45 in this cohort indicating stable behavioural
performance. Notably, while checking for behavioural stability, a sig-
nificant group difference began to emerge with our key dependent
variable of interest—percent premature responses. As such, upon
reaching stability, animals received one further 5CSRTT session before
being euthanised for tissue collection. Average data from sessions
44–46 are presented, corresponding to those that occurred in the 12th
week after the start of lithium treatment. Additional analyses of
W.K. Adams, et al.
premature responses from sessions 38–46 were conducted as described
in the next section.
2.4. Determining subgroups for brain tissue analyses
We aimed to identify any changes in inflammatory markers most
strongly associated with the lithium-induced reduction in premature
responding. To this end, samples were isolated from a subgroup of rats
that were most responsive to lithium’s anti-impulsive effects, as ob-
served over the last two weeks of testing during which a main effect of
lithium treatment was observed (i.e. sessions 38–46). The percentage
change in premature responding was calculated for each rat as follows,
with ‘final’ referring to mean % premature responses from sessions
44–46, and ‘initial’ referring to those from sessions 38–40: 100×(final-
initial)/initial. Percentage change scores were ranked for each group to
determine the 6 most lithium-responsive animals (i.e. those with the
greatest percentage decrease in impulsive responding), and the 6 most
stable control animals (i.e. those with percentage change scores around
the group median). Our goal was to enhance the signal-to-noise ratio of
our central cytokine assay, and minimise the impact of neurobiological
factors associated with changes in impulsivity that were driven by
causes other than lithium treatment. Blood samples from the full cohort
were subject to lithium analysis and cytokine discovery assays, whereas
brain samples were analysed for the subgroup of lithium-responders
and stable controls only.
2.5. Blood and brain tissue collection
Approximately 24 h after the last 5CSRTT session (< 24 h post-
feeding), trunk blood and brain tissue samples were collected following
rapid decapitation. The order in which samples were collected alter-
nated between lithium-treated and control rats to minimise any time of
day effects. Blood was immediately aliquoted into collection tubes for
the separation of serum or plasma (BD Microtainer® Tubes: SST
#365967 and K2 EDTA #365974, BD Diagnostics, Franklin Lakes, NJ,
USA) and placed on ice. Tubes were later centrifuged for 10 min (4500
g, 4 °C). Plasma aliquots were stored at −80 °C until use, whereas
serum aliquots were promptly analysed for lithium levels. Tissue pun-
ches from key brain regions implicated in the regulation of premature
responding on the 5CSRTT, namely the medial prefrontal cortex
(mPFC), orbitofrontal cortex (OFC), and nucleus accumbens (NAc)
(Dalley et al., 2004; Winstanley et al., 2006), were immediately wet-
dissected on a cold surface and frozen on dry ice. Brain samples were
stored at –80 °C until homogenised for cytokine assays.
2.6. Serum lithium analysis
On the day of tissue collection, serum aliquots from all animals were
delivered to the UBC Hospital (Rapid Response Laboratory, Vancouver,
BC, Canada) for the clinical determination of lithium concentrations
using flame atomic absorption spectrophotometry.
2.7. Multiplex cytokine assays
To assist in identifying molecular targets for brain tissue analysis,
plasma samples for all animals were outsourced for multiplex cytokine
assays (Rat 23-Plex Cytokine “Discovery Assay”; Eve Technologies,
Calgary, AB, Canada). All samples were run in duplicate. Six of the 23
cytokines assayed could not be reliably quantified (IL-5, IL-17, GM-CSF,
G-CSF, TNF-α, VEGF), whereas concentrations were obtained for the
following 17 analytes: IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12(p70),
IL-13, IL-18, MCP-1, MIP-1α, RANTES, Eotaxin, GRO/KC, IP-10, IFN-γ,
and Leptin. Plasma cytokine levels are reported as pg/mL.
Brain samples from the mPFC, OFC and NAc from the subgroups of
lithium-responsive and control rats were prepared for in-house cytokine
assays. Samples were defrosted in ice-cold lysis buffer (150 mM NaCl,
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20 mM Tris pH 7.5, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100), which
was prepared fresh with the addition of 1 complete mini protease in-
hibitor cocktail tablet (Roche Diagnostics, Indianapolis, IN, USA),
200 µL phosphatase inhibitors 2 and 3 (200 µL each; Sigma-Aldrich, St.
Louis, MO, USA), 100 µL 1 M NaF, and 40 µL PMSF (from 500 mM stock
in DMSO) per 10 mL volume. Samples were homogenised by ultra-
sonication and centrifuged for 15 min (16,110 g, 4 °C). Aliquots of
supernatant were removed for cytokine analyses and stored at –20 °C.
The remaining supernatant was subject to protein quantification using
the Pierce BCA Protein Assay Kit (#23227, Thermo Fisher Scientific,
Waltham, MA, USA), according to the manufacturer’s protocol. To ac-
count for limited volumes, samples were diluted 1:20 in working re-
agent and run in duplicate; values were averaged to give the total
protein content (mg/mL).
Brain homogenates were then subject to two electro-
chemiluminescent immunoassays. Levels of IL-1β, IL-4, IL-6, IL-10 and
TNF-α were simultaneously quantified using a custom 5-plex kit
(#K153A0H-2, Meso Scale Discovery, Rockville, MD, USA). RANTES
levels were detected using a customised single-plex assay, in which
capture antibodies from commercially-available antibody pairs (#900-
K72, Peprotech, Rocky Hill, NJ, USA) were printed in the wells of
standard-bind multiplex plates (#N45ZA-1 proprietary printing service,
Meso Scale Discovery); RANTES detection antibodies were derivatised
with electrochemiluminescent SULFO-Tag NHS-ester by the manu-
facturer, and 2.88 µL SULFO-TAG labeled Streptavidin was added with
the detection antibody. Samples were diluted 1:2 in diluent 42.
Immunoassays were otherwise run according to the manufacturer’s
standard protocols. Plates were read using a Sector Imager 2400 (Meso
Scale Discovery) and data were analysed using the Discovery
Workbench software v. 4.0 (Meso Scale Discovery). The lower limit of
detection (LLOD) for the assays varied by plate and analyte. The fol-
lowing LLOD ranges were observed (pg/mL): IL-1β, 3.65–5.40; IL-4,
0.0525–0.0682; IL-6, 3.04–4.57; IL-10, 0.281–0.705; TNF-α,
0.0933–0.164. The LLOD for RANTES was 0.0869 pg/ml. Tissue levels
were adjusted and reported as pg (cytokine)/mg of protein.
2.8. Statistical analyses
All behavioural and molecular data were assessed by ANOVA using
Systat 13 (Systat Software, Inc., Chicago, IL, USA), with the between-
subjects factor Group (2 levels: Control, Lithium), and repeated-mea-
sures where appropriate.
To determine whether rats differed with respect to weight gain as a
result of chronic lithium treatment, body weights (g) over the course of
the experiment were analysed with the within-subjects factor Time (14
levels: 14 experimental weeks, including ‘acclimation’ and
t = 0–12 weeks’ food-restriction/lithium diet).
5CSRTT variables were analysed separately, including the percen-
tage of trials in which correct, omitted or premature responses were
made; the total number of perseverative responses or trials completed;
and latencies for correct responses and reward collection, as described
previously (Adams et al., 2017; Adams et al., 2015). Changes in im-
pulsive responding over the testing period were analysed during data
collection using a rolling three-session bin. For simplicity, data are
presented here with the within-subjects variable ‘session block’ (3 le-
vels: mean responses over sessions 38–40, 41–43, and 44–46), for
comparison with % change scores used to determine subgroups.
5CSRTT variables expressed as percentages were arcsine transformed to
offset the influence of an artificially-imposed ceiling (i.e. 100%).
Cytokine data were analysed separately, with outlying data points
identified in the initial ANOVAs removed from analysis. All data were
log transformed to meet normality assumptions; untransformed data
are presented for clarity. All data are expressed as mean ± Standard
Error of the Mean (SEM). Differences were considered significant where
p < 0.05; differences where 0.05 < p < 0.1 are reported.
W.K. Adams, et al.
3. Results
3.1. Serum lithium levels and body weight
In serum samples obtained within 24 h of ingesting the last dose,
rats maintained on the lithium-supplemented diet showed circulating
lithium levels that fell within the therapeutic range for humans
(Fig. 1a). Average concentrations in lithium-treated rats were
0.78 ± 0.08 mM/L compared to negligible levels in controls
(0.01 ± 0.01 mM/L; Group: F(1,27) = 113.1, p < 0.001). Non-zero
lithium levels in samples from two control rats likely resulted from
cross-contamination during blood collection/processing.
Chronic lithium treatment led to significantly reduced body weight
over the course of the experiment (Fig. 1b; Group: F(1,27) = 44.6,
p < 0.001; Time: F(13,351) = 83.0, p < 0.001; Group × Time:
F(13,351) = 30.2, p < 0.001). Body weights did not differ between
groups in the free-feeding acclimation week nor after 2 weeks of food-
restriction, which included the first week of lithium exposure (Fig. 1b;
Acclimation, week ‘0′ and week ‘1′: all F-values ≤ 2.1, p-va-
lues ≥ 0.163). From the second week of lithium exposure until the end
of the experiment, lithium-treated rats weighed markedly less than
controls (Fig. 1b; weeks ‘2-12′: all F-values between 32.5 and 85.6, all
p-values < 0.001). Visual consideration of these data suggests that
lithium-treated rats lost more weight than controls in the second and
third week of lithium exposure, after which the rate of body weight gain
tended to parallel that of control animals.
3.2. 5CSRTT
After 12 weeks of chronic lithium treatment, rats showed reduced
rates of premature responding compared to controls, reflecting reduced
levels of motor impulsivity (Fig. 2a; 3a; Group- s38-40: F(1,27) = 3.14,
p = 0.09; s41-43: F(1,27) = 5.17, p = 0.031; s44-46: F(1,27 = 6.7,
p = 0.015). None of the other 5CSRTT behavioural variables were af-
fected by chronic lithium treatment, including the accuracy of stimulus
detection, response omissions or latencies, reward collection latencies,
perseverative responding and the total number of trials completed
(Fig. 2b-g; all F-values ≤ 1.2, p-values ≥ 0.285). Importantly, the
observation that response omissions and reward collection latencies
were unchanged in lithium-treated animals indicates that the reduction
in premature responses did not arise from an overall state of disen-
gagement with the task, or a general reduction in motor activity.
Further examination of impulsive responding during the testing
period was conducted to compare this behaviour in the subgroups of
rats selected as candidates for brain cytokine assays (subgroup A) to the
remaining animals (subgroup B) and the full experimental cohort.
Within subgroup A, premature responding was comparable across
groups in the first session block (s38-40), but significantly declined over
the subsequent sessions in lithium-treated animals (Fig. 3b,c; Group:
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Brain, Behavior, and Immunity 89 (2020) 339–349
Fig. 1. Chronic lithium administration via custom-
formulated chow leads to clinically-relevant serum
lithium levels and reduced body weight in healthy
male rats. Graphs show a) serum lithium con-
centrations obtained at the end of the experiment
(< 24 h after last dose) and b) weekly body weights
for control (n = 16) and lithium-treated (n = 13)
rats. ‘A’: free-feeding acclimation week; weeks
‘0–12′: food-restricted. Data are shown as
mean ± SEM. *p < 0.001 compared to controls.
F(1,10) = 1.3, p = 0.287; Session block: F(2,20) = 4.2, p = 0.030;
Group × Session block: F(2,20) = 5.6, p = 0.019). A very similar pat-
tern was observed when lithium-treated animals from subgroup A were
compared to controls from subgroup B (Group: F(1, 14) = 0.51,
p = 0.486; Session block: F(2, 28) = 3.87, p = 0.033, Group × Session
block: F(2, 28) = 4.3, p = 0.024), and the two subgroups of control rats
also did not differ across this time period (Group: F(1, 14) = 0.56,
p = 0.465; Session block: F(2, 28) = 1.12, p = 0.345, Group × Session
block: F(2, 28) = 0.1, p = 0.991). The control rats selected for the brain
analyses were therefore largely representative of the whole cohort.
Interestingly, the percentage change scores—used to isolate the
subset of most lithium-responsive rats—were not different between li-
thium-treated and control groups when data from the full cohort were
analysed (Fig. 3c; Group: F(1,27) = 0.4, p = 0.513), suggesting a sig-
nificant proportion of lithium-treated rats were not showing a decrease
in premature responding over this time period. As is clear from Fig. 3b,
these animals (subgroup B lithium) appeared to be showing consistently
low levels of motor impulsivity compared to control subjects, perhaps
indicative of a more pronounced response to lithium treatment, rather
than lithium resistance. Further a posteriori analyses across rolling three
session bins instead revealed that subgroup B lithium-treated rats
(n = 7) made significantly fewer premature responses while the task
was still being acquired (Fig. 3b: session 21–23, Group- all lithium-
treated rats vs controls: F(1, 27) = 2.53, p = 0.12, - lithium subgroup B
vs controls: F(1,21) = 5.77, p = 0.026; session 21–46, Group- all li-
thium-treated rats vs controls: F(1, 27) = 2.78, p = 0.11, - lithium
subgroup B vs controls: F(1, 21) = 7.96, p = 0.01). This posed some-
thing of a conundrum. It is well-documented that levels of premature
responding can vary considerably within healthy cohorts of rats, and
this can be associated with a range of neurobiological differences (e.g.
Belin et al., 2008; Caprioli et al., 2015; Dalley et al., 2007; Fernando
et al., 2012). Given the early emergence of this change, either these
animals were significantly more responsive to lithium, or they were
naturally less impulsive, and our experimental design did not permit us
from determining which was most likely. By combining behavioural
analyses with concurrent lithium treatment, our goal was to identify
lithium-induced changes in cytokine expression that were most likely
affecting impulse control by analysing brain samples at the same time-
point at which impulsivity significantly declined. Out of an abundance
of caution, we therefore focused our analyses on the brain tissue har-
vested from the six rats in subgroup A which showed the most robust
decline in premature responding in the time period immediately prior
to tissue collection. Serum lithium levels and patterns of body weight
changes in subgroup A were also comparable to that shown by the full
experimental cohort (Supplementary Figure S1). In addition, beha-
vioural performance for the other 5CSRTT variables by the subgroups
matched that of the full cohort (Supplementary Figure S2).
W.K. Adams, et al.
Fig. 2. Chronic lithium administration reduces motor impulsivity in healthy male rats after 12 weeks. Graphs show average behavioural responses in the 5CSRTT
over the final test sessions (44–46) for control (n = 16) and lithium-treated (n = 13) rats. Chronic lithium treatment selectively reduced levels of a) motor
impulsivity, without affecting b) attentional accuracy, c) omissions, d) response or e) reward collection latencies, f) perseverative responding or g) total trials
completed. Data are shown as mean ± SEM. *p = 0.015 compared to controls.
3.3. Plasma cytokines
Plasma samples from all animals were outsourced for a multiplex
cytokine assay to narrow down the list of potential immune markers for
brain tissue analysis. This “discovery assay” revealed that chronic li-
thium administration reduced circulating levels of IL-1β, IL-10,
RANTES and leptin in all lithium-treated rats compared to controls
(Table1; Group: IL-1β, F(1,27) = 9.2, p = 0.005; IL-10, F(1,27) = 10.7,
p = 0.003; RANTES, F(1,27) = 4.6, p = 0.041; Leptin, F(1,25) = 12.9,
p = 0.001). The full complement of lithium-treated rats also tended to
have reduced circulating levels of IL-2 (Table 1; Group: F(1,26) = 3.9,
p = 0.058). Plasma levels of these cytokines did not differ between
subgroups A and B (all Fs ≤ 2.224, all p’s ≥ 0.15). Within subgroup A,
differences between lithium and control-treated rats failed to reach
statistical significance, likely due to the lower statistical power re-
sulting from the smaller sample size (all Fs ≤ 4.47, all p’s ≥ 0.061).
Although the following cytokines and chemokines could be detected,
chronic lithium treatment was not found to alter their plasma levels
under the present experimental conditions: IL-1α, IL-4, IL-6, IL-
12(p70), IL-13, IL-18, MCP-1, MIP-1α, Eotaxin, GRO/KC, IP-10, IFN-γ
(Table 1; all F-values ≤ 2.8, p-values ≥ 0.105).
3.4. Brain cytokines
We therefore explored if any corresponding changes in IL-1β, IL-10
and RANTES levels in brain regions involved in impulse control were
associated with the anti-impulsive effects of chronic lithium treatment,
using tissue samples from subgroup A. IL-4, IL-6 and TNF-α were also
determined since these immune markers have been implicated in BD
and suicidality, and in the mechanism of action of lithium (Maddu and
Raghavendra, 2015; Munkholm et al., 2013; Munkholm et al., 2015).
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Brain, Behavior, and Immunity 89 (2020) 339–349
Leptin was not included, however, as reduced circulating leptin levels
likely resulted from body weight losses in lithium-treated rats, and
previous work in our laboratory indicates that leptin does not influence
premature responding in the 5CSRTT (Adams et al., 2018; Adams et al.,
2015)
Results from in-house cytokine assays of OFC, mPFC and NAc
samples from the subgroups of lithium-responsive and stable control
animals are shown in Fig. 4; the OFC was the only region in which
differences in immune marker content were revealed. Compared to
controls, the subgroup of lithium-responsive rats tended to show re-
duced levels of IL-1β, as well as significantly reduced levels of IL-6 and
RANTES, in the OFC (Fig. 4a,c,f; OFC–Group: IL-1β, F(1,10) = 3.7,
p = 0.082; IL-6, F(1,9) = 7.2, p = 0.025; RANTES, F(1,9) = 6.6,
p = 0.030). In the mPFC or NAc, tissue levels of these markers were
similar between the subgroups (Fig. 4a,c,f; mPFC, NAc–Group: IL-1β, all
F-values ≤ 1.9, p-values ≥ 0.198; IL-6, all F-values ≤ 1.5, p-va-
lues ≥ 0.249; RANTES, all F-values ≤ 0.1, p-values ≥ 0.723). Ad-
ditionally, there were no significant differences in the levels of IL-4, IL-
10 and TNF–α in the brain regions examined between lithium-re-
sponsive rats and controls (Fig. 4b,d,e; all regions–Group: IL-4, all F-
values ≤ 1.2, p-values ≥ 0.302; IL-10, all F-values ≤ 1.5, p-va-
lues ≥ 0.251; TNF–α, all F-values ≤ 2.0, p-values ≥ 0.190).
4. Discussion
Here we show that chronic, dietary administration of lithium for
12 weeks selectively reduced a measure of motor impulsivity in rats, in
keeping with the therapeutic use of lithium to treat BD. Plasma levels of
the immune-signalling molecules IL-1β, IL-10 and RANTES were se-
lectively reduced in lithium-treated animals at this time-point. Ex vivo
analyses of brain tissue taken from a subgroup of animals showing a
W.K. Adams, et al. Brain, Behavior, and Immunity 89 (2020) 339–349

Fig. 3. Change in impulsive responding over

the 5CSRTT testing period in all lithium-
treated and control animals compared to the
subgroups used for brain cytokine analysis.
Panels a and b show the average behavioural
responses in the 5CSRTT over all test ses-
sions. The shaded area contains data obtained
during acquisition of the task. A significant
difference between lithium-treated (n = 13)
and control rats (n = 16) emerged over ses-
sions 41–46 (panel a). When evaluating the
degree to which a decrease in premature re-
sponding was observed in each lithium-
treated rat over this time (panel c), it became
clear that some animals (Lithium-A, n = 6)
showed a significant shift, whereas others
(Lithium-B, n = 7) did not. A posteriori ana-
lyses show that the null effect in subgroup B
occurs because these animals had been
making significantly fewer premature re-
sponses since session 21 (panel b). Data are
shown as mean ± SEM. *p < 0.05,
***p < 0.001, compared to controls.

reduction in premature responding at the same time confirmed reduced regional specificity in lithium’s actions on immune signalling. These
levels of IL-1β (trend-level significance) and RANTES selectively within findings add to the growing body of literature suggesting an association
the OFC, as well as a local reduction in IL-6. No changes in cytokine between immune signalling in the frontal cortex and the expression of
levels were detected in the mPFC or NAc, suggesting some degree of impulsive behaviors (Vonder Haar et al., 2019; Vonder Haar et al.,

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W.K. Adams, et al. Brain, Behavior, and Immunity 89 (2020) 339–349

Table 1
Plasma cytokine levels in both the full cohort of control and lithium-treated rats, and in the subgroup used for brain cytokine analyses. * denotes group difference
relative to control at p < 0.05, ** at p ≤ 0.005, *** at p ≤ 0.001. # denotes a trend.
Control-All rats Lithium-All rats Control-Subgroup A Lithium-Subgroup A

IL-1α 109.5 ± 35.8 92.1 ± 23.2 119.1 ± 81.5 60.3 ± 28.0

IL-1β 7.7 ± 2.5 1.3 ± 0.9** 2.2 ± 1.1 0.8 ± 0.5
IL-2 22.4 ± 13.0 26.2 ± 6.7# 45.4 ± 39.6 20.0 ± 3.6
IL-4 9.5 ± 2.1 13.0 ± 2.1 10.7 ± 4.5 9.6 ± 2.7
IL-6 128.2 ± 39.6 157.7 ± 42.3 158.0 ± 99.5 103.5 ± 36.0
IL-10 198.0 ± 61.7 30.1 ± 16.9** 71.5 ± 36.8 14.9 ± 2.7#
IL-12(p70) 15.8 ± 2.4 18.2 ± 2.3 15.7 ± 5.4 15.8 ± 3.2
IL-13 56.2 ± 11.3 50.6 ± 11.2 46.2 ± 17.6 33.1 ± 13.4
IL-18 21.0 ± 3.9 13.2 ± 1.3 30.8 ± 10.3 11.6 ± 0.6
MCP-1 (CCL2) 99.3 ± 12.2 87.4 ± 10.4 69.5 ± 14.6 79.7 ± 5.0
MIP-1α (CCL3) 4.8 ± 0.6 4.0 ± 0.5 5.5 ± 1.4 4.0 ± 0.8
RANTES (CCL5) 28575.2 ± 7234.4 10407.4 ± 3857.7* 22670.5 ± 7068.3 11010.9 ± 6143.6
Eotaxin (CCL11) 6.9 ± 1.6 9.6 ± 2.5 3.7 ± 1.9 7.4 ± 2.2
GRO/KC (CXCL1) 318.7 ± 37.9 247.4 ± 39.9 277.7 ± 64.6 235.0 ± 51.9
IP-10 (CXCL10) 8.3 ± 1.5 9.1 ± 1.2 7.3 ± 2.4 8.0 ± 1.1
IFN-γ 18.1 ± 4.3 26.9 ± 5.5 21.8 ± 11.1 16.0 ± 3.9
Leptin 5161.0 ± 273.1 3293.1 ± 400.5*** 4780.2 ± 465.3 3087.0 ± 709.0#

Data are expressed as mean pg/mL ± SEM for control (n = 15–16) and lithium-treated (n = 11–13) rats. *p < 0.05, **p < 0.01, ***p ≤ 0.001, #p = 0.058
compared to controls.

2016; Vonder Haar et al., 2017).
Indeed, this is not the first time that either plasma or brain levels of
these cytokines have been associated with impulsivity. Of 23 cytokines
analysed in plasma through multiplex immunoassay, similar to the
approach taken here, RANTES was the only analyte to be both sig-
nificantly elevated in a group of male alcohol-dependent subjects and
also positively correlated with levels of impulsivity (Manzardo et al.,
2016). Elevated RANTES levels have previously been associated with
other psychiatric disorders including BD, schizophrenia, and major
depression (Oglodek et al., 2014; Stuart and Baune, 2014). Numerous
studies have also reported positive correlations between plasma levels
of IL-6 and impulsive behaviours (Coccaro et al., 2014; Isung et al.,
2014; Lindqvist et al., 2009; Munkholm et al., 2013; Sutin et al., 2010),
and elevated cerebrospinal fluid levels of IL-1β have been associated
with manic episodes in BD (Soderlund et al., 2011). As such, decreasing
central and peripheral levels of these cytokines would be expected to
track a decrease in impulsive behaviour, as observed here in response to
lithium administration.
Lithium was administered over many weeks in the diet, rather than
via injection, to avoid the feeling of malaise induced by this substance.
Indeed, pairing food rewards with injections of lithium chloride is a
highly effective way to devalue those foods or induce conditioned
aversion (O'Donnell and Gould, 2007). Animals were food-restricted
throughout behavioural testing, and rats generally consumed all the
diet placed in their home cages, yet animals in the lithium-treated
group consistently weighed less than rats fed the control diet. This
seemed to emerge within the first 2–3 weeks of diet exposure, before
5CSRTT testing began, during which time the rats may have been ac-
climating to the ingestion or taste of lithium. After this period, the rate
of weight gain across both groups was comparable. Although not ideal,
the difference in weights across the cohorts is unlikely to explain the
reduction in premature responding observed in the lithium-treated
group; we have shown that even pathologically obese rats show intact
baseline rates of premature responding in the 5CSRTT (Adams et al.,
2018). If anything, reduced consumption of chow in the home cage
should increase motivation for the sugar pellet rewards available during
the 5CSRTT, resulting in elevated premature responding, trials com-
pleted, accuracy of performance, and reduced omissions and response
latencies (Robbins, 2002). Lithium-treated rats showed none of these
behavioural effects. A previous report suggests that bolus lithium ad-
ministration can also decrease motor activity in rats, raising the con-
cern that the reduction in premature responding could simply reflect
general motor slowing (Cappeliez and White, 1981). The fact that
dietary lithium treatment did not concomitantly increase omissions or
response latencies, or decrease the total trials completed, argues
strongly against this conclusion.
Rats were also pair-housed for the duration of the experiment and
were not isolated during food consumption. It is therefore possible that
one animal in each cage consumed more or less of the allotted food.
However, the weight of cage-mates remained comparable over time,
and serum lithium levels were remarkably homogenous, suggesting that
dyads shared the available food relatively equitably. We also conducted
pilot experiments prior to initiating this behavioural study to ensure
that feeding animals in pairs resulted in similar serum levels of lithium
as when animals were fed in isolation (Levesque, 2013). Indeed, dosing
rats with 14 g of 2.91 g/kg lithium diet resulted in serum lithium levels
within the therapeutic range used to treat mood disorders
(0.5–1.5 mM/L). Although it is notoriously difficult to equate doses
across species due to differences in drug metabolism, the similarity in
serum levels and reduction in impulsive responses in both humans and
rats supports the hypothesis that the behavioural and physiological
changes observed in the rodent may be informative regarding the
beneficial effects of lithium in clinical populations.
To optimize the translational relevance of these findings, it would
have been useful to assess cytokine changes in a validated animal model
of BD or mania (Alda, 2015). Unfortunately, the underlying genetic or
neurobiological cause of BD remains unknown, rendering generation of
such an animal model difficult. Nevertheless, inhibition or knockdown
of the dopamine transporter, deletions of mitochondrial DNA, and
mutations of one of the key genes regulating circadian rhythms in mice
are able to recapitulate some of the core symptoms (Kato, 2017;
Kristensen et al., 2018; van Enkhuizen et al., 2015). Determining
whether lithium likewise reduces premature responding in these
models, and decreases pro-inflammatory cytokine expression in re-
levant brain circuits, would be a valuable next step. These models lar-
gely rely on the generation of transgenic mice, which makes the im-
plementation of complex operant behavioural testing paradigms more
challenging, but not impossible (e.g. van Enkhuizen et al., 2013).
However, the study of behavioural expression of impulsivity in other-
wise-healthy rats using well-validated tasks like the 5CSRTT has gen-
erated a wealth of knowledge regarding the neurobiological substrates
underlying impulse control in both healthy and diseased states (Dalley
et al., 2004; Pattij and Vanderschuren, 2008; Winstanley, 2011). Li-
thium may also have beneficial effects in reducing impulsivity and re-
lated behaviours (suicide, aggression) in healthy individuals, with
particularly prominent effects in males (Amado et al., 2005; Ishii et al.,

345
W.K. Adams, et al.
2015; Sheard et al., 1976; Shiotsuki et al., 2016). Determining the
mechanism through which chronic lithium decreases premature re-
sponding in this paradigm is therefore still a worthy goal. It will also be
important to determine whether lithium is similarly effective in female
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Brain, Behavior, and Immunity 89 (2020) 339–349
Fig. 4. Reduced levels of pro-inflammatory markers in the orbitofrontal cortex
are associated with reduced motor impulsivity following chronic lithium
treatment. Graphs show tissue content of a) IL-1β, b) IL-4, c) IL-6, d) IL-10, e)
TNF-α, and f) RANTES in the orbitofrontal and medial prefrontal cortices, and
nucleus accumbens (OFC, mPFC, NAc), of the subgroups of lithium-treated rats
and controls used for molecular analyses (n = 5–6 per group). Lithium-treated
rats that showed the greatest percentage decrease in premature responding in
the 5CSRTT showed reduced levels of a) IL-1β, c) IL-6 and f) RANTES in the
OFC. Data are expressed as mean ± SEM. *p < 0.05, #p = 0.082 compared
to controls.
animals. Although the clinical response to lithium in BD, and the in-
cidence of the disorder itself, is generally consistent across sex (Diflorio
and Jones, 2010; Viguera et al., 2000), there are marked differences in
the immune system across sex which may be particularly important to
the neuroimmune actions of lithium (Rainville and Hodes, 2019).
Logistical and financial constraints notwithstanding, we acknowl-
edge it would have been better to analyse brain samples from all li-
thium-treated rats, rather than just those from a subgroup, and that this
latter approach reflects a major limitation of the current study. As
summarised in the results, an argument could also be made that sub-
group B showed an earlier and more sustained response to lithium
treatment, and that brain samples from these animals should instead
have been prioritised for cytokine analysis. Rather than being re-
presentative of the response to lithium treatment in general, the brain
cytokine levels collected here from subgroup A may instead reflect
those of “late responders”. Alternatively, we could have selected a
subgroup of animals based on plasma cytokine response to lithium.
However, the major added benefit of combining behavioural testing
with lithium treatment was the possibility of identifying candidate
changes in cytokine levels, among the myriad possible, that were time-
locked with a reduction in impulsivity. By adhering to our original
experimental design, we may have therefore missed larger or more
widespread changes in cytokine expression present in the remaining
lithium-treated rats. Follow-up studies will be essential to substantiate
these effects, and determine if they replicate across multiple cohorts.
In the current study, chronic dietary lithium administration de-
creased circulating levels of IL-10, which is an anti-inflammatory cy-
tokine. This seems to contrast with the consensus view that lithium
treatment increases IL-10 production (see Nassar and Azab, 2014).
Perhaps notably, most of the studies supporting this conclusion have
used shorter durations of lithium treatment than used here. Relatively
acute administration (1–5 days) of lithium in vivo, or incubation of cells
isolated from whole blood cultures with lithium in vitro, lead to in-
creased basal production of IL-10 in samples from healthy volunteers
(Agrawal et al., 2013; Coant et al., 2011; Liu et al., 2011; Maes et al.,
1999; Rapaport and Manji, 2001; Szuster-Ciesielska et al., 2003). Acute
administration of lithium in mice in vivo also increased IL-10 produced
in response to an infectious agent (Tay et al., 2012; Zhang et al., 2009),
whereas incubating cultured rat cortical glia cells with lithium likewise
increased IL-10 levels in response to lipopolysaccharide challenge
(Green and Nolan, 2012). However, there is a relative paucity of studies
documenting the effects of chronic lithium administration on IL-10
expression in either humans or rats. Three months of chronic lithium
treatment in BD patients decreased the number of peripheral blood
lymphocytes producing IL-10 (Boufidou et al., 2004), whereas serum
levels of IL-10 were unfortunately undetectable after four weeks of li-
thium treatment in healthy volunteers and rapid-cycling bipolar pa-
tients (Rapaport et al., 1999). Nevertheless, given that lithium acts to
inhibit GSK3β, we would have predicted a rise in circulating IL-10.
Future studies will hopefully confirm whether the reduction in IL-10
observed here after 12 weeks of daily lithium treatment is reproducible
or an anomaly. Alternatively, a reduction in IL-10 following such long-
term administration may be secondary to reduced levels of pro-in-
flammatory cytokines.
Although we observed lower levels of IL-6 in the OFC of lithium-
W.K. Adams, et al.
treated rats, we could not detect any group differences in circulating IL-
6. This is reminiscent of other studies in humans in which plasma and
CSF levels of IL-6 do not correlate (Isung et al., 2014; Lindqvist et al.,
2009), and therefore do not specifically invalidate the significance of
the OFC finding. The fact that changes in RANTES, IL-1β and IL-6 were
restricted to the OFC and were not seen in the other brain regions
sampled, is also consistent with previous demonstrations that cytokine
expression varies significantly across area, both at baseline and in re-
sponse to different challenges (Bastos et al., 2018; Hayley et al., 2013;
Huang et al., 2008; Song et al., 2006). The behavioural response to local
up- or down-regulation of cytokine signalling also shows dramatic re-
gional selectivity, which may be related to differences in neuronal ac-
tivity (Hong et al., 2016; Schafer et al., 2012; Song et al., 2006). Given
the central role of the OFC in BD, the response to lithium, and the
regulation of impulsivity in preclinical models, it may therefore not be
surprising that the effects of lithium on cytokine levels were most
pronounced here. Nevertheless, although the changes we observed in
peripheral and central cytokine levels following lithium treatment are
consistent with a role for immune signaling molecules in mediating
lithium’s therapeutic effects, these data remain correlational. Demon-
strating that local infusions of IL-6, IL-1β, or RANTES/CCL5 impair
premature responding, or that preventing signalling through their re-
spective receptors can improve impulse control, would be critical to
proving that these cytokines play a causal role in regulating impulsivity
within the OFC.
Such experiments are not trivial to conduct because relatively little
is known regarding the mechanism or time course through which these
proteins act, either in concert or independently, to alter brain activity.
In the peripheral immune system, RANTES/CCL5 promotes inflamma-
tion through the recruitment of T cells, eosinophils, basophils and the
activation of natural-killer cells. IL-6 is an important factor in the
transition from innate to acquired immunity, inducing maturation of B-
cells into antibody-producing cells (Jones, 2005), whereas IL-1β is one
of the major drivers of the systemic response to infection, triggering the
recruitment of numerous other pro-inflammatory cytokines. However,
the roles these cytokines may play in maintaining healthy brain func-
tion in the absence of immune challenges is much less well-docu-
mented. RNA-Seq analyses indicate that microglia are the most likely
source of cytokine synthesis in the CNS, followed by astrocytes and
oligodendrocytes (Zhang et al., 2014). These data suggest changes in
cytokine levels likely alter neuronal function through microglial inter-
actions. mRNA for RANTES/CCL5, IL-1β, and IL-6 has been found in
rodent cortical neurons (Bandtlow et al., 1990; Lanfranco et al., 2017;
Ma et al., 2018; Ringheim et al., 1995), raising the possibility that
neurons themselves may contribute to cytokine signalling. However,
when the neuronal and glial cell responses to stimulation of the immune
system via the endotoxin lipopolysaccharide have been directly com-
pared, increases in IL-6 mRNA are much more pronounced in glial cells
(e.g. Ma et al., 2018; Qian et al., 2014), in keeping with the consensus
view that microglia mainly produce cytokines in the brain, at least in
response to major immune challenges.
While the current data cannot provide any insight into the me-
chanism through which cytokine levels may modulate cognitive func-
tion, previous studies suggest some compelling possibilities. Microglia
play a fundamental role in synapse formation and elimination, not just
during development, but also in the healthy adult brain (Parkhurst
et al., 2013; Trapp et al., 2007; Wake et al., 2009). Chemical signalling
between neurons and glial cells, particularly astrocytes, can also en-
hance or depress synaptic activity through gliotransmission (Gundersen
et al., 2015). It is therefore possible that changes in microglial or as-
trocytic regulation of synaptic plasticity in the OFC could result in
improved cognitive control and reduced impulsivity, although this re-
mains to be empirically determined.
Cytokines can also alter neuronal function through binding to
neuronally-expressed cytokine receptors and activating janus-asso-
ciated kinase/signal transducer and activator of transcription (Jak/
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Brain, Behavior, and Immunity 89 (2020) 339–349
STAT) pathways (Donegan et al., 2015; Qian et al., 2014). These Jak/
STAT pathways have been critically implicated in synaptic plasticity
within neurons (Nicolas et al., 2012; Yirmiya and Goshen, 2011). In-
deed, numerous studies suggest that IL-6 in particular could play a
critical role in the homeostatic control of learning and memory (see
Erta et al., 2012; Gruol, 2015 for review). Of perhaps particular re-
levance to the current findings, local inhibition of either IL-6 or Jak/
STAT activity within the rat OFC had similar effects on reversal lear-
ning—a form of cognitive flexibility—as a serotonin 5-HT2A receptor
antagonist, and in vitro data suggest that IL-6 may exert these effects
through directly modulating 5-HT2 receptor signalling (Donegan et al.,
2014; Donegan et al., 2015; Furr et al., 2012). 5-HT2A antagonism, and
theoretically by extension attenuating IL-6 activity, improves impulse
control on the 5CSRTT (Higgins et al., 2003; Winstanley et al., 2003a;
Winstanley et al., 2003b), similar to the effects of lithium reported here.
The serotonergic system has also been implicated in the pro-impulsive
effects of IL-1β (Hassanain et al., 2005; Hassanain et al., 2003;
Howerton et al., 2014). Although currently speculative, such work
suggests mechanisms through which the lithium-induced decreases in
orbitofrontal cytokine levels may feasibly decrease premature re-
sponding.
In sum, we have shown here that chronic lithium treatment can
selectively reduce motor impulsivity, and that this behavioural change
is accompanied by reductions in certain pro-inflammatory cytokine
levels in the orbitofrontal cortex, a key brain region involved in reg-
ulating this type of impulsive response. Although the link between cy-
tokine levels and behaviour reported here is purely correlational, nu-
merous studies indicate that these cytokines can impact neuronal
function, leading to the generation of testable hypotheses whereby local
expression of these proteins may contribute to lithium’s beneficial ef-
fects. Collectively, our findings therefore support the hypothesis that
lithium attenuates impulsivity through an anti-inflammatory me-
chanism. Improved understanding of how immune signalling molecules
alter cognition may offer hope for a new generation of therapeutics
targeting neuro-immune interactions in psychiatric disorders.
Acknowledgements
This work was supported by a grant awarded to CAW from the Coast
Capital Savings Depression Research Fund through the UBC Institute of
Mental Health. CAW received salary support through the Michael Smith
Foundation for Health Research and the Canadian Institutes of Health
Research New Investigator Program during this time. WKA was sup-
ported by philanthropic donations provided by Paul and Diane Erickson
and the Gillespie Family. In the past three years, CAW has consulted for
Shire, and been retained as an expert witness by Hogan Lovells LLP, on
unrelated matters, and received due compensation. The authors con-
firm they have no other conflicts of interest or financial disclosures to
make. We gratefully acknowledge Prof. Joanne Weinberg and Prof.
Timothy J. Kieffer for access to their resources to conduct cytokine
assays.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.bbi.2020.07.018.
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