S566
ations in mice. A 14 mg/ml dose of clozapine did not show
any changes in sensorimotor gating, while 3.5 and 7 mg/ml
dose of clozapine significantly increased the PPI. Thus, this
dosage range can be used in future studies requiring chronic
icv delivery of clozapine.
References
[1] Horacek, J., Bubenikova-Valesova, V., Kopecek, M.,
Palenicek, T., Dockery, C., Mohr, P., Höschl, C., 2006.
Mechanism of action of atypical antipsychotic drugs and
the neurobiology of schizophrenia. CNS drugs 20 (5), 389–409.
[2] Pardridge, W., 2012. Drug transport across the blood-brain bar-
rier. J Cereb Blood Flow Metab 32 (11), 1959–1972.
[3] Raja, M., 2011. Clozapine safety, 35 years later. Curr Drug Saf
Jul 6 (3), 164–184.
doi: 10.1016/j.euroneuro.2018.11.839
P.824 PPP1r1b gene and others in the dorsal striatum
of mice with movement disturbances
D. Smagin ∗, A.G. Galyamina, I.L. Kovalenko,
N.N. Kudryavtseva, V.N. Babenko
The Federal Research Center Institute of Cytology and Ge-
netics, Siberian Branch of Russian Academy of Sciences,
Novosibirsk, Russia
Background: Dopaminergic neurons play critical role in mul-
tiple brain functions and have been implicated in several
neurologic and psychiatric disorders. Nigrostriatal dopamin-
ergic system is the main player in the regulation of motor
activity. One well-known target for the actions of dopamine
is DARPP32 which is expressed by Ppp1r1b gene. This protein
is critical for dopamine dependent striatal synaptic plastic-
ity. The paper aimed to study the differentially expressed
dopaminergic genes in male mice with differences in psy-
chomotor activity. In particular, we addressed the Ppp1r1b
observed gene network context along with causes and con-
sequences of ultrahigh Ppp1r1b gene expression in the stria-
tum. Another task was to investigate the regulatory molecu-
lar mechanisms/factors governing Ppp1r1b expression rate.
Methods: Disturbances in motor behavior in male mice
are induced by positive and negative social experience
in daily agonistic interactions [1,2]. Three groups were
studied: depressive males who were defeated after 21 days
and displayed hypoactivity (low locomotion, immobility,
depressiveness); aggressive males with repeated positive
fighting experiences during 21 days which demonstrated
hyperactivity and stereotypic behaviors; the controls – mice
without consecutive experiences of agonistic interactions.
RNA-Seq data was obtained from the collected samples
of the dorsal striatum using standard NGS protocol. The
Cufflinks program was used to estimate the gene expression
levels in FPKM units. We also used supervised gene clus-
tering to assess genes expression co-variaton rate in the
targeted genes subsets.
Results: In the analysis of molecular changes in the dorsal
striatum the Ppp1r1b gene expression was hundred times
greater in the striatum than in other brain regions. In
depressive mice, decreased expression of the dopaminergic
Abstracts
Сomt, Drd1, Drd2, Ppp1r1b and Snca genes and enhanced
expression of the Sncb, and Slc6a3 were shown. In aggres-
sive mice increased Drd4 gene and decreased Th gene were
found. The highly expressed genes contain strong regulation
related ctcf binding sites and CpG islands. Based on our
data gene expression profiles we observed highly correlated
cluster of striatonigral genes network (11 genes), centered
on Drd1 dopamine receptor gene, and a separate expanded
cluster of striatopallidal Drd2 mediated gene network (38
genes) including Ppp1r1b gene. It was ascertained via GO
annotation that striatonigral network genes identified are
responsible for calcium signaling. The striatopallidal gene
cluster is enriched for genes involved in locomotor be-
havior. Given the Ppp1r1b gene maintains the outstanding
expression within the network, it’s quite plausible it drives
the expression of connected genes in a coordinated manner.
Conclusion: In the analysis of molecular changes in the
dorsal striatum, the Ppp1r1b gene expression was hundred
times greater in the striatum than in other brain regions.
It has been suggested that this gene is specific and can
play a key role in regulating neurochemical changes and,
as a consequence, contribute to an understanding of the
possible mechanisms of impaired motor activity in various
diseases, accompanied by changes in dopaminergic activity.
It is believed that, as a target for dopamine, Ppp1r1b gene
can serve as a therapeutic site in the treatment of many
psychoneurological disorders.
Disclosure statement: Supported by Russian Science Foun-
dation (grant No 14-15-00063) and Budget project (No
0324-2018-0016).
References
[1] Kudryavtseva, N.N., Smagin, D.A., Kovalenko, I.L., Vishnivet-
skaya, G.B., 2014. Repeated positive fighting experience in
male inbred mice. Nat Prot 9 (11), 2705–2717.
[2] Smagin, D.A., Galyamina, A.G., Kovalenko, I.L., Babenko, V.N.,
Tamkovich, N.V., Borisov, S.A., Tolstikova, T.G., Kudryavt-
seva, N.N., 2018. Altered expression of neurotransmitter genes
in the dorsal striatum of male mice with psychomotor distur-
bances. Zh Vyssh Nerv Deiat Im I P Pavlova 68 (2) in press.
doi: 10.1016/j.euroneuro.2018.11.840
P.825 Involvement of GABAA receptor gamma 2 sub-
unit in the anxiety-like behaviour and cognitive dys-
function in pentylenetetrazole-kindled rats
Z. Gáll 1,∗, K. Kelemen 2, S. Koncz 1, I.I. Toth 1, I. Mihály 2,
M. Kolcsár 1
1
University of Medicine and Pharmacy of Tirgu Mures, Phar-
macology and Clinical Pharmacy, Tirgu Mures, Romania
2
University of Medicine and Pharmacy of Tirgu Mures, De-
partment of Physiology, Tirgu Mures, Romania
Objective: Anxiety and cognitive dysfunction frequently
remain untreated comorbidity of epilepsy [1]. GABAA
receptor positive allosteric modulators are non-specific
anxiolytic agents, and neither as anticonvulsants are not
efficient. Moreover, these drugs induce cognitive dysfunc-
Abstracts
tion in patients [2]. Subtype-selective modulators can
be synthesized, which may produce effective anxiolysis
without sedation [3]. This study aimed to investigate the in-
volvement of GABAergic inhibition in anxiety-like behavior
and cognitive function in pentylenetetrazole (PTZ)-kindled
rats. The expression of GABAA receptor gamma2 subunit
was studied along with behavioral changes during the
PTZ-kindling and in the fully-kindled state.
Methods: Male adult Wistar rats (n = 12) underwent win-
PTZ kindling protocol [4]. The control group (n = 8) was
administered saline injections. Animals were tested twice
(in the latent period and after the kindling procedure)
on a battery of behavioral tests including open-field (OF),
elevated plus maze (EPM), marble burying (MB), and radial-
arm maze (RAM) tests. In each test, the anxiety-related
behaviors were monitored and analyzed using Noldus Etho-
Vision XT software. At the end of the experiment, the brain
of the animals was fixed in 4 % paraformaldehyde. Double
immunofluorescence staining was performed on 60 μm thick
brain slices using anti-GABAAgamma2 antibody and DAPI.
Images were acquired by Leica TCS-SP8 confocal laser
scanning microscope and analyzed using a semiquantitative
fluorescence imaging technique.
Results: The histological analysis revealed a significant de-
crease of GABAA receptor gamma2 subunit expression in the
stratum oriens and radiatum of CA1 subregion of the hip-
pocampus in case of the kindled group compared to the con-
trol (p<0.001). A similar pattern of changes was found in the
hippocampal CA3 region, however, no differences between
the subregions were observed. The locomotor activity of
the animals in the OF did not show any significant difference
neither between the groups nor between the trials. The
time spent in the center zone decreased at the second test-
ing in case of both groups. In the EPM, the number of entries
and the time spent in the open-arm did not show a signifi-
cant difference between the groups at any trial. In the MB
test, the fully-kindled animals showed an increased activity
and interacted more with marbles than in the latent period.
The number of marbles buried or dislocated, the number of
entries and time spent in the zone delimited by marbles sig-
nificantly increased (F(1,42) = 7.736, p<0.05), but the kin-
dling did not significantly modify marble burying compared
to controls. In the RAM test, all animals learned to collect
the baits. The total number of errors did not differ signif-
icantly between the groups. Neither the working memory
nor the reference memory of the animals did not decrease.
Conclusion: The reduction of GABAA receptor gamma2
subunit expression in the kindled animals, and the loss of
GABAergic inhibition correlates with a decreased seizure
threshold and increased severity of seizures, but the
anxiety-state of the kindled animals or their cognitive per-
formance did not differ from the controls. So, the observed
decrease of GABAA receptor gamma2 subunit expression
did not cause any behavioral changes, which suggests that
PTZ-kindling model of epilepsy do not reflect the human
psychiatric comorbidities of epilepsy.
References
[1] Mula, M., 2013. Treatment of anxiety disorders in epilepsy: an
evidence-based approach. Epilepsia 54, 13–18.
S567
[2] Jembrek, M.J., Vlainic, J., 2015. GABA receptors: pharmaco-
logical potential and pitfalls. Curr Pharm Des 21, 4943–4959.
[3] Atack, J.R., 2011. GABAA receptor subtype-selective modula-
tors. I. α2/α3-selective agonists as non-sedating anxiolytics.
Curr Top Med Chem 11, 1176–1202.
[4] Davoudi, M., Shojaei, A., Palizvan, M.R., Javan, M., Mirnajafi-
Zadeh, J., 2013. Comparison between standard protocol and a
novel window protocol for induction of pentylenetetrazol kin-
dled seizures in the rat. Epilepsy Res 106, 54–63.
doi: 10.1016/j.euroneuro.2018.11.841
P.827 Does physical activity associated with chronic
food restriction alleviate anxiety like behavior?
M. Méquinion 1, S. Eddarkaoui 2, D. Blum 2, J. Vignau 3,
V. Tolle 4, O. Viltart 4,∗
1
Monash University, Dept Physiology-, Clayton- Melbourne,
Australia
2
INSERM, UMR-S 1172 JPArc, Lille, France
3
CHRU, Psychiatry, Lille, France
4
INSERM, U894- Center of Psychiatry and Neuroscience,
Paris, France
Background: Anorexia nervosa (AN) is an eating disorder
characterized by excessive weight loss, persistent food
restriction and inappropriate physical activity in regard
to their energy homeostasis. This disease affects mostly
the young women. Despite its complex etiology, metabolic
and endocrine aspects are important to consider as key
pathophysiological determinants of AN. The comorbidity
with depression and/or anxiety disorders might contribute
to the “chronification” of the disease [1].
Objectives: We aim here to investigate in a rodent
model mimicking numerous physiological AN alterations
[2] whether a) chronic food restriction is associated with
anxiety-like behavior and b) physical activity plays a role in
regulating the level of anxiety.
Methods: For this purpose we first exposed young female
mice (2 per cage) to a chronic food restriction (FR) com-
bined or not with access to a running wheel (W). The mice
(n = 6/group) were compared to a group of mice fed ad
libitum without (AL) or with a wheel running activity (ALW).
We tested the anxious-like behavior of all these mice by
submitting them after two weeks of protocol to the fol-
lowing tests: hyponeophagia, marble burying test, elevated
plus maze (EPM), open field (OF), light and dark box. To be
in similar feeding conditions, all mice received the same
amount of food the evening before the test (1g) and one
hour before the beginning of the test (1g). Then the last
day, mice were submitted to the restraint test during 30
min to measure the HPA axis reactivity. All tests have been
performed during one week between 8:00 to 11:00 am.
Plasma corticosterone concentration was measured with
an ELISA kit (AssaysPro). Data were analyzed using ANOVA
followed by a Fisher post-hoc test.
Results: FRW and FR mice showed a stable 25 % decrease
of the body weight. The behavior of FRW mice was similar
to those of AL and ALW mice in the OF and the EPM while
FR mice showed reduced of anxious-like behavior in these
tests. The FRW mice displayed the lowest latency to reach