Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice
Aim: To study the effect of new vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and Lewis lung carcinoma metastasis in the anxious adult male mice of C57Bl/6J strain. This type of receptors was thought to act as potential targets mediating the effect of negative psychoemotio...
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nasplib_isofts_kiev_ua-123456789-1386602025-02-23T18:58:23Z Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice Amikishieva, A.V. Ilnitskaya, S.I. Nikolin, V.P. Popova, N.A. Original contributions Aim: To study the effect of new vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and Lewis lung carcinoma metastasis in the anxious adult male mice of C57Bl/6J strain. This type of receptors was thought to act as potential targets mediating the effect of negative psychoemotional state on tumor progression. Methods: Anxiety-like psychoemotional state of the animals was produced using chronic social conflict model. Used behavioral tests were elevated plus-maze, social interaction test and open field test. Tumor cells were administrated on background of double or sixfold SSR149415 injections and the number of metastases in the lung were calculated 17 days later. Results: SSR149415 reduced the anxiety-like behavior measured in the elevated plus-maze and social interaction tests and did not affect locomotor activity in the open field test. Double and sixfold administration of the compound to such mice before and after inoculation of the tumor cells produced no effect on the metastasis rate. Conclusion: vasopressin V1b receptor is involved in the mediation of anxious behavior of animals but is not involved in the mechanism underlying the influence of negative psychoemotional state on Lewis lung carcinoma metastasis. The authors are grateful to Dr. Claudine SerradeilLe Gal (Sanofi Eventis Recherche & Developpement, France) for kind provision of V1b receptor antagonist. 2011 Article Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice / A.V. Amikishieva, S.I. Ilnitskaya, V.P. Nikolin, N.A. Popova // Experimental Oncology. — 2011. — Т. 33, № 3. — С. 126-129. — Бібліогр.: 25 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/138660 en Experimental Oncology application/pdf Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Original contributions Original contributions Amikishieva, A.V. Ilnitskaya, S.I. Nikolin, V.P. Popova, N.A. Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice Experimental Oncology |
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Aim: To study the effect of new vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and Lewis lung carcinoma metastasis in the anxious adult male mice of C57Bl/6J strain. This type of receptors was thought to act as potential targets mediating the effect of negative psychoemotional state on tumor progression. Methods: Anxiety-like psychoemotional state of the animals was produced using chronic social conflict model. Used behavioral tests were elevated plus-maze, social interaction test and open field test. Tumor cells were administrated on background of double or sixfold SSR149415 injections and the number of metastases in the lung were calculated 17 days later. Results: SSR149415 reduced the anxiety-like behavior measured in the elevated plus-maze and social interaction tests and did not affect locomotor activity in the open field test. Double and sixfold administration of the compound to such mice before and after inoculation of the tumor cells produced no effect on the metastasis rate. Conclusion: vasopressin V1b receptor is involved in the mediation of anxious behavior of animals but is not involved in the mechanism underlying the influence of negative psychoemotional state on Lewis lung carcinoma metastasis. |
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Article |
| author |
Amikishieva, A.V. Ilnitskaya, S.I. Nikolin, V.P. Popova, N.A. |
| author_facet |
Amikishieva, A.V. Ilnitskaya, S.I. Nikolin, V.P. Popova, N.A. |
| author_sort |
Amikishieva, A.V. |
| title |
Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice |
| title_short |
Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice |
| title_full |
Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice |
| title_fullStr |
Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice |
| title_full_unstemmed |
Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice |
| title_sort |
effect of vasopressin v1b receptor antagonist, ssr149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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2011 |
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Original contributions |
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| citation_txt |
Effect of vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and lewis lung carcinoma metastasis in mice / A.V. Amikishieva, S.I. Ilnitskaya, V.P. Nikolin, N.A. Popova // Experimental Oncology. — 2011. — Т. 33, № 3. — С. 126-129. — Бібліогр.: 25 назв. — англ. |
| series |
Experimental Oncology |
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| fulltext |
126 Experimental Oncology 33, 126–129, 2011 (September)
EffEct of VasoprEssin V1b rEcEptor antagonist,
ssr149415, on anxiEty-LikE bEhaVior and LEwis Lung
carcinoma mEtastasis in micE
A.V. Amikishieva*, S.I. Ilnitskaya, V.P. Nikolin, N.A. Popova
Institute of Cytology and Genetics of Siberian Department of Russian Academy of Sciences, Novosibirsk,
Russian Federation 630090
Aim: To study the effect of new vasopressin V1b receptor antagonist, SSR149415, on anxiety-like behavior and Lewis lung carci-
noma metastasis in the anxious adult male mice of C57Bl/6J strain. This type of receptors was thought to act as potential targets
mediating the effect of negative psychoemotional state on tumor progression. Methods: Anxiety-like psychoemotional state of the
animals was produced using chronic social conflict model. Used behavioral tests were elevated plus-maze, social interaction test and
open field test. Tumor cells were administrated on background of double or sixfold SSR149415 injections and the number of me-
tastases in the lung were calculated 17 days later. Results: SSR149415 reduced the anxiety-like behavior measured in the elevated
plus-maze and social interaction tests and did not affect locomotor activity in the open field test. Double and sixfold administration
of the compound to such mice before and after inoculation of the tumor cells produced no effect on the metastasis rate. Conclusion:
vasopressin V1b receptor is involved in the mediation of anxious behavior of animals but is not involved in the mechanism underlying
the influence of negative psychoemotional state on Lewis lung carcinoma metastasis.
Key Words: anxiety, Lewis lung carcinoma, metastasis, vasopressin receptor, SSR149415
The effect of psychoemotional state on tumor pro-
gression is an established phenomenon although its
underlying mechanisms remain elusive. Three decades
ago it was demonstrated that rearing of animals in over-
crowded conditions and subsequent activation of the
stress system of organism can significantly change the
incidence of tumors and the number of tumor metastatic
nodules [22]. The study of two types of transplantable
tumors carried out in mice showed that animals with
anxiety-like behavior induced by rearing in stressful
conditions have twice as much of experimental me-
tastases as intact mice [10, 16], while anxiolytic drug
diazepam reduces the number of metastases [9].
Hypothalamic hormone vasopressin potentiating
the action of corticoliberin can increase the secretion
of the adrenocorticotropic hormone and subserves
significantly to the formation of negative stress-
induced psychoemotional states [13, 23]. Distribution
of vasopressinergic fibers and receptors in the limbic
structures of the brain suggests the involvement of this
neuropeptide in emotional responses to stress [11].
The animals with depression semiotics and patients
with major depression were shown to have an in-
creased plasmatic level of vasopressin. In both cases
administration of antidepressants normalizes the
hormone level [4, 12]. Rats selected for high anxiety
behavior exhibit innate elevated expression of vaso-
pressin mRNA and hypothalamic receptor binding [18].
Exogenous intraventricularly administered vasopres-
sin produces anxiogenic effect while its antagonists
alleviate the symptoms of anxiety [2, 7]. Clinically ef-
fective anxiolytic drugs reduce the expression of the
vasopressin precursor gene [23].
In the experiments with Lewis lung carcinoma (LLC)
metastasis, administration of vasopressin was shown
to increase the number of metastatic nodules in the
lungs of mice [1], similarly as it occurs in animals with
an impaired psychoemotional state. Still it is so far not
clearly how the mechanisms of vasopressin action
on metastasis and psychoemotional state interrelate.
Physiological action of vasopressin is known
to be mediated by receptors of three types: V1a, V1b,
and V2. The receptors of the former two types are
widespread in the CNS structures [8, 20] and are in-
volved in anxious behavior [5]. Receptors of V1a type
are also present on unstriped muscles of the brain
vessels and V1b receptors — on corticotroph pituitary
cells to implement the effect of vasopressin on the
stress axis of organism [21]. Receptors of both types
are sensitive to emotional stress. Anxiety-like behav-
ior in animals correlates with enhanced V1-receptor
binding in the CNS [12]. Addition of antisense mRNA
of V1a receptor to mice results in a pronounced allevia-
tion of anxiety-like behavior and impairment of social
recognition. Chronic immobilization stress increases
V1a mRNA level in rat brain [17].
Recently synthesized specific vasopressin V1b-
receptor antagonist, SSR 149415 [7] provides the
opportunity for distinguishing the contribution of re-
ceptors of this type to increasing of tumor metastasis
in animals with pronounced anxiety-like behavior,
which has become the purpose of this study.
matEriaLs and mEthods
Animals. Male C56B1/6J strain mice, weigh-
ing 25–28 g, housed in standard conditions of the
vivarium of the Institute of Cytology and Genetics,
Siberian Branch of the Russian Academy of Sciences
Received: May 23, 2011.
*Correspondence: E-mail - amik@bionet.nsc.ru
Fax - 7 383 333 12 78
Abbreviations used: CNS – central nervous system; EPM - el-
evated plus-maze; LLC – Lewis lung carcinoma.
Exp Oncol 2011
33, 3, 126–129
Experimental Oncology 33, 126–129, 2011 (September) 127
were used. All experimental procedures with animals
were carried out in accordance with the international
rules established by the European Community Council
Directive (86/609/EEC).
Anxious pattern of mice behavior was produced
using a 10-day chronic social conflict model [15].
Behavior Evaluation Methods
The elevated plus-maze (EPM). This method
is used to measure anxiety-like behavior of the mice
placed in unfamiliar environment [24]. The four-minute
experimental sessions were recorded with video
camera. The conventional behavioral measures were
recorded with the use of the “Mouse” software pack-
age (Institute of Cytology and Genetics, SB RAS). The
most significant anxiety-related measures included
time spent in the central area of the maze, time in the
closed arms and time in the open arms and area (sum
of central area and open arms times), and latent period
of closed-arm entries. Exploratory behavior measures
were the number of dipping postures with head below
the level of the maze and stretching postures.
The “Partition” social interaction test. This test
is used to obtain quantitative measures of sociability
(communicativeness) of the animals relative to the
level of anxiety [6]. The behavior of the mice in anxiety-
like state was assessed in response to a known aggres-
sive male and a strange male placed by turns in the
neighboring compartment of the home cage across
a perforated transparent partition. During the 6-minute
test periods (3+3), the number of approaches and time
at the partition were recorded for the test male and the
derived measure — average time at the partition per
approach — was calculated as an indirect measure
of communicative motivation of the mice.
Open field test. This test is used to evaluate lo-
comotor and exploratory activities. Three-minute test
sessions were recorded with video camera. The be-
havior of the mice was recorded and calculations were
made to find the distance, average speed, immobility
time, number of upright postures and explorations
of the setting holes as well as the time in the central
and peripheral zones of the setting.
Oncological Experiments. Lewis lung carcinoma
cells were inoculated into the tail vein (0.5 ml of sus-
pension, 170 thousands cells/mouse). Seventeen days
later the mice were decapitated; the lungs were fixed
in 10% formalin solution. The number of metastatic
nodules was calculated under a microscope with 8-fold
magnification. Two experiments were performed:
double administration of SSR149415 — a day be-
fore and on the day of the tumor cell inoculation;
sixfold administration of SSR149415 — once a day
during 6 days. Tumor cells were inoculated on the
fourth day of SSR149415 injections.
Pharmacology. Vasopressin V1b receptor an-
tagonist, SSR149415, (10 mg/kg, suspended in 0.6%
methylcellulose and 0.5% Tween solution) was admin-
istrated once 1 hour before the behavioral tests. Also,
in the day of tumor cell inoculation, the compound was
administrated 1 hour before LLC cell injection too. Since
the drug is known to be equally active at intraperitoneal
and oral administration [7], both methods were used
in our experiments. The vehicle in equal volumes was
administered by the same methods to control mice.
Statistical Processing. The results were analyzed
with the use of Student’s t-criterion using STATISTIKA
6.0 software package. Each experimental group con-
sisted of 11–15 animals.
rEsuLts
Behavioral tests
Elevated plus-maze. Intraperitoneal adminis-
tration of SSR149415 significantly reduced the level
of anxiety-like behavior in animals (Table 1). The mice
spent almost twice as less time in the EPM closed arms
and thrice as much time in the open areas as compared
to control. The latent period for entries into the closed
arms also grew significantly. The time of staying in the
center of the maze increased 2.5 times versus the con-
trol, which could be interpreted as a reduced speed of
“decision making” on the backdrop of the drug action.
Exploratory activity measures — dipping postures with
heads below the maze and stretching postures showed
no reliable difference from the control values mainly
due to a considerable scattering of data.
Table 1. Effect of vasopressin receptor antagonist, SSR149415, on the be-
havior of male mice with anxiety-like behavior in elevated plus-maze test
Anxious animals
+ vehicle (15)
Anxious animals +
SSR149415 (13)
Time in closed arms, sec 195.53 ± 13.96 109.92 ± 25.11 **
Latent period of closed arm entry, sec 19.20 ± 7.31 78.46 ± 25.50 *
Time in open arms, sec 5.60 ± 2.73 29.69 ± 18.06
Time in the center of the maze, sec 38.00 ± 13.34 97.77 ± 22.90 *
Time in open areas, sec 43.60 ± 13.76 127.46 ± 25.85 **
Number of dipping postures 2.87 ± 0.82 5.46 ±1.14
Number of stretching postures 0.73 ± 0.41 2.69 ± 1.00
Note. *р<0.05; **p<0.01 compared to animals with anxiety, administra-
tion of the vehicle.
“Partition” social interaction test. Oral adminis-
tration of SSR149415 produced no effect on the main
measure of communicativeness — time spent by the
mice at the partition exploring the smell of a male
placed in the neighboring compartment (Table 2).
Nonetheless, the drug administration to male mice
with anxiety-like symptoms in the situation with unfa-
miliar male behind the partition doubled the derived
measure — the time at the partition per approach —
as compared to control mice suggesting an enhance-
ment in mice motivation for communication with
a conspecific and reduction of anxiety-like behavior
following the administration of SSR149415.
Table 2. Effect of vasopressin receptor antagonist, SSR149415, on the be-
havior of male mice with anxiety-like behavior in the social interaction test
Anxious animals
+ vehicle (15)
Anxious animals +
SSR149415 (11)
Familiar male:
Number of interactions (N)
Time of interactions (T), sec
Average interaction time (T/N), sec
4.23 ± 0.43 3.90 ± 0.85
41.31 ± 7.73 38.70 ± 6.12
9.69 ± 1.59 12.44 ±2.30
Unfamiliar male:
Number of interactions 4.13 ± 0.75 3.09 ± 0.77
Time of interactions, sec 67.47 ± 12.17 69.73 ± 10.79
Average interaction time, sec 17.37 ± 1.72 34.76 ± 8.15 *
Note. *р<0.05; **p<0.01 compared to animals with anxiety-like behavior
receiving the vehicle.
128 Experimental Oncology 33, 126–129, 2011 (September)
Open field test. Oral administration of SSR149415 did
not affect locomotor or exploratory activity of animals
with anxiety-like behavior in the test (Table 3).
Table 3. Effect of vasopressin receptor antagonist, SSR149415, on the
behavior of male mice with anxiety-like behavior in open field test
Anxious animals
+ vehicle (15)
Anxious animals +
SSR149415 (11)
Passage distance, cm 290 ± 37 308 ± 42
Immobility time, sec 49.3 ±10.4 62.4 ± 9.6
Speed (distance/time in motion, cm/sec) 2.2 ± 0.2 2.5 ±0.3
Time in the central area, sec 50.2 ± 10.2 36.0 ± 7.4
Time in peripheral area, sec 130.8 ± 10.1 142.2 ±7.6
Number of holes explored 6.4 ± 1.1 5.3 ± 1.1
Number of vertical postures 3.1 ± 0.7 2.7 ± 0.7
metastasis
As a rule, the number of LLC metastatic nodules
in the lungs positively correlates with the level of anxiety
in animals. In our experiments this correlation was reli-
ably reproduced in the second experiment (Table 4).
Table 4. Effect of vasopressin receptor antagonist, SSR149415, on me-
tastasis of Lewis lung carcinoma in the lungs of male mice with anxiety-
like behavior
Number of meta-
static nodules
Experiment 1: Intact animals + vehicle 9.9 ± 1.7 (14)
Animals with anxiety-like behavior + vehicle 12.8 ± 1.34 (15)
Animals with anxiety-like behavior + SSR149415 14.54 ± 1.93 (13)
Experiment 2: Intact animals + vehicle 8.2 ± 1.3 (11)
Animals with anxiety-like behavior + vehicle 14.7 ± 2.2 (15) *
Animals with anxiety-like behavior + SSR149415 15.0 ± 2.2 (14)
Note *р<0.05 as compared with intact animals receiving the vehicle.
In the first experiment, after double intraperitoneal
administration of SSR149415 to mice with anxiety-like
behavior the number of metastatic nodules did not
change as compared with administration of vehicle
to control group.
In the second experiment, after six oral administra-
tions of SSR149415 the number of metastatic nodules
did not differ from control values.
discussion
Possible influence of emotional state of an indi-
vidual on the course of tumor progression has been
of interest exclusively for practicing psychologists
[3, 25]. In the literature under discussion is whether
and to what degree psychotherapy can influence the
treatment of patients with tumors. The opinions on this
topic seem to be equally divided. We failed to find any
biochemical studies aiming to elicit the correlation
between psychoemotional state of patients (includ-
ing the activity of hypothalamus-pituitary-andrenal
axis and vasopressinergic systems) and the efficacy
of anticancer therapy.
The studies of the last decade demonstrated that
vasopressin is a principal regulator of emotional be-
havior. Studies in humans support the co-activation
of the central vasopressinergic system at anxiety
and depression disorders [23] while clinically effec-
tive anxiolytic drugs and antidepressants reduce the
expression of the vasopressin gene in the CNS [4].
It is vasopressin that provokes hyperactivation of the
hypothalamus-pituitary-adrenal axis under chronic
emotional stress by acting on pituitary corticotrophs
through V1b receptors [14].
The leading role of vasopressin in behavioral
phenomena of anxiety and depression and in stress-
induced diseases was also demonstrated on animal
models [18]. The Brattleboro rates deficient for vaso-
pressin gene demonstrate elements of depression-like
behavior [19]. In our experiment with mice a single
subcutaneous administration of vasopressin caused
behavioral reactivity of animals in the open field test [1].
In our experiments anxiety-like behavior was
formed during 10 days by means of daily 10-minute
agonistic interactions with a stronger male who was
kept during the rest of the day in the neighboring
compartment of the cage behind a perforated trans-
parent partition. This procedure is sufficient to form
a stable anxiety-like state in experimental animals
[15]. Administration of V1b receptor antagonist,
SSR149415, resulted in a visible reduction of anxiety
level in the elevated cross-maze test and an increase
of motivation in animals with anxiety-like behavior for
social interaction with a strange male in the partition
test. Therefore, in our experiments we reproduced the
results confirming the anxiolytic effect of the drug [7].
After injection of vasopressin antagonist no changes
in locomotor activity were recorded.
Tumor cells were inoculated 1–2 days after the last
fighting. Administration of SSR149415 was made in the
1st experiment a day before and tumor cell administra-
tion day; in the 2nd experiment — 3 days before, during
carcinoma cells inoculation day and two next days.
The hours and days immediately after inoculation are
thought to be a sensitive period for tumor cells dis-
semination and future growth of metastatic nodules.
During this period, double and sixfold administration
of SSR149415 did not change the increased intensity
of Lewis lung carcinoma cells metastasis in animals
with anxiety-like behavior irrespective of the route
of drug administration. These results provide more
assurance to claim that the previously found enhanc-
ing effect of a single vasopressin administration on the
process of malignant metastasis is mediated not
by V1b receptors. In other words, the V1b receptors
are not involved in tumor cell dissemination and growth
and the hormone action is not due to its psychotropic
behaviorally inhibiting effect.
In this situation V1a receptors preserve their appeal
to researchers as possible mediators of vasopressin
effect on metastasis. Receptors of this type are located
in different regions of the brain and blood vessels and
together with V1b receptors are involved in behavioral
phenomena [20]. The latter site of action of the vaso-
pressinergic hormone can influence dissemination
of tumor cells by luminal narrowing of lungs vessels
under the effect of vasopressin.
acknowLEdgmEnts
The authors are grateful to Dr. Claudine Serradeil-
Le Gal (Sanofi Eventis Recherche & Developpement,
France) for kind provision of V1b receptor antagonist.
Experimental Oncology 33, 126–129, 2011 (September) 129
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