ISSN 1062-3590, Biology Bulletin, 2017, Vol. 44, No. 10, pp. 1252–1256. © Pleiades Publishing, Inc., 2017.

Original Russian Text © M.V. Kropotkina, E.V. Kuznetsova, N.Yu. Feoktistova, 2016, published in Povolzhskii Ekologicheskii Zhurnal, 2016, No. 3, pp. 263–270.

Seasonal Changes in the Hormonal Response

of Male Eversmann’s Hamsters (Allocricetulus eversmanni,
Cricetinae, Rodentia) to Olfactory Signals from Conspecific Females
M. V. Kropotkina*, E. V. Kuznetsova, and N. Yu. Feoktistova
Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences, Moscow, 119071 Russia
*e-mail: marriyashka@yandex.ru
Received June 11, 2016

Abstract—Reproductive success demands both the male and female reaching a particular physiological sta-
tus, which is usually stimulated in rodents by certain olfactory signals. The hormonal response of male Evers-
mann’s hamster (Allocricetulus eversmanni) to the natural excretions (urine, midventral gland secretion
(MVGS) of conspecific females was studied during several seasons of the year, with the animals kept in con-
ditions of natural lighting and temperature. In the fall, no reliable changes in the testosterone level in males
in response to any signal presented were noted. In the winter and spring, a reliable (P < 0.05) increase in the
testosterone level was caused by the female MVGS odor. In the summer, a similar reaction only to conspecific
female urine (P < 0.05) was noted. Moreover, in the summer and fall, males showed a reliable decrease in the
cortisol level in response to the odor of female urine (P < 0.05). In the winter and spring, a similar result was
caused by exposure to both odors. The progesterone level in males reliably (P < 0.05) increased upon exposure
to both female urine and MVGS in all seasons of the year, except for the female MVGS in the summer. There-
fore, the decreased cortisol level in combination with the simultaneously increased testosterone and proges-
terone levels allow the males of the species under study to achieve their maximum reproductive success,
despite the high degree of their intraspecific aggression.

Keywords: Allocricetulus eversmanni, urine, midventral gland secretion, testosterone, progesterone, cortisol
DOI: 10.1134/S1062359017100077

INTRODUCTION not with the animal itself (Rozhnov, 2011). Restric-

The olfactory analyzer is phylogenetically the most
ancient sensory system (Wilson, 1970; Tamar, 1976). It
has now been established that the genes encoding olfac-
tory receptors account for up to 5% of the total genome
of the vertebrate, which makes the superfamily of genes
of olfactory receptors one of the largest and demon-
strates their exceptional importance for mammals-
macrosmatics (Buck, 2004). Chemical signals can
inform about population density and, accordingly, reg-
ulate of both territorial relations and the reproductive
process (Shilov, 1977, 2002). Odor signals are excep-
tionally important for nocturnal rodents, especially
those solitary, as they allow animals to avoid unwanted
contact with other animals, but provide necessary
information from the scent marks of other individuals.
The use of chemical signals has both positive and
negative aspects. The advantages of chemical commu-
nication for nocturnal animals are determined by the
fact that chemical signals are easily perceived by recip-
ients in the dark and are energetically cheap, because,
as natural metabolites are often used, they are retained
for a long time in the external environment, allowing
individuals to have contact only with the mark, and
tions are also imposed by the physical properties of the
environment, the distance from the sender to the
recipient, the chemical arsenal of the sender, the phys-
iological parameters of the signal formation, etc.
(Sachs, 1999). However, with respect to the majority
of representatives of the subfamily Cricetinae, the pos-
itive aspects of chemical communication clearly pre-
vail over the negative ones due to the fact that direct
contacts in the populations of these animals are often
associated with aggressive interactions.
Chemical signals regulate the production of sex ste-
roids through the main and additional olfactory sys-
tems. Testosterone and its metabolites increase sexual
motivation and stimulate spermatogenesis and sexual
behavior (Wilson, 1999). Progesterone is a precursor of
the corticosteroids and testosterone, at the same time
suppressing the synthesis of the latter (intramuscular
injection) (Jeyaraj et al., 2001). Finally, chemical sig-
nals of females cause a whole chain of biochemical and
hormonal reactions, stimulating courtship and ensuring
successful mating (Kotenkova, 2014).
The chemical composition of olfactory signals and
their perception depend on the hormonal state of the

1252
 SEASONAL CHANGES IN THE HORMONAL RESPONSE
donor individuals and recipients, which, in turn,
depends on the duration of the photoperiod (season of
year) (Zucker et al., 1980; Powers et al., 1985; Ferkin
et al., 1994). For instance, during the spring and sum-
mer adult male and female produce odors that are
attractive to opposite-sex conspecifics. A high level of
testosterone in seasonally breeding animals provokes
the interest of males in the smell of females and the
development of sexual behavior, while its low levels
eliminate interest and suppress sexual behavior (Bron-
son, 1988, 1989). Thus, the role of chemical signals
during the year may be different, and the nature of the
response to scent signals can determine the mutual
relations between individuals (Zucker et al., 1980; Fer-
kin et al., 1994; Leonard and Ferkin, 1999; Feokti-
stova, 2008; Kropotkina, 2012). The hormonal male
responses to the females odor signals allows make
assumptions about the nature of the relationship
between individuals of different sexes in the different
seasons of the year. It is especially important for spe-
cies that are difficult to observe in nature. In particu-
lar, a representative of the genus Allocricetulus—Evers-
mann’s hamster—is an example of such a species and
it is widely distributed: from the Middle and Lower
Volga to the Irtysh River in the east and to northern
Xinjiang in the south (Sludskii et al., 1977). However,
there is few data on its biology. There are contradic-
tory reports that the species hibernates in the Trans-
Ural region, whereas in the Lower Volga region, it is
active and reproduces in winter (Shchepot’ev, 1959;
Flint et al., 1970; Vorontsov, 1982).
Eversmann’s hamster has never been kept in labo-
ratory until now. However, there are a few works
devoted to its behavior in nature in the spring and
summer (Ryurikov et al., 2003, 2005). It has been
shown, for example, that adult animals occupy indi-
vidual home range from 0.1 to 7.3 hectares. Individual
home range of females do not overlap with each other,
but the males cover, as a rule, the sections of several
females and intersect with other males. Each individ-
ual uses several burrows, with one burrow used for
resting by several individuals alternately.
During breeding, adult males actively search for
receptive females (Ryurikov et al., 2005), while both
males and females leave scent marks both from urine
and from rubbing the MVGS onto the substrate (our
observations).
At the Severtsov Institute of Ecology and Evolu-
tion, Russian Academy of Sciences, Eversmann’s
hamster was bred in laboratory conditions. It was
established that in the fall–winter period, it is charac-
terized by an irregular hypothermia with a drop in
temperature to 5–6°C and bouts of sleep of no more
than two days (Ushakova et al., 2010; Klevezal’ et al.,
2015). However, not all individuals demonstrate win-
ter hypothermia: some remain active, which confirms
the possibility of winter reproduction in this species.
BIOLOGY BULLETIN Vol. 44 No. 10 2017
1253
Owing to this, it seems important to identify the
features of perception of excretions and hormonal
responses of male Eversmann’s hamsters to olfactory
signals from female conspecifics in different seasons of
the year when kept under natural light and tempera-
ture conditions.
MATERIALS AND METHODS
Ten males from the first generation of Eversmann’s
hamsters caught in the Saratov Trans-Volga region
and constituting the “Collection of Live Mammalian
Wild Species of Rodents and Insectivores” of the
Severtsov Institute of Ecology and Evolution, Russian
Academy of Sciences, were used in the experiment.
The animals were kept singly, in an unheated room,
protected from rain and snow, in cells measuring 21 ×
16 × 14 cm, in natural light and temperatures condi-
tions (Moscow). Cotton wool was used as the nesting
material, and wood shavings were used as a substrate.
As the diet of Eversmann’s hamster in nature is rich in
protein (Flint and Golovkin, 1961), in addition to
grain fodder and fresh vegetables, the animals were fed
meat, cottage cheese, and eggs.
On the same day, only one excretion or water
(control exposure) were presented to the recipient
animals, the experiment was conducted no more
often than once a week. Excretion were obtained
from at least two donors (in order to minimize the
effect of individual odor characteristics) and stored
in a refrigerator (2–4°C) for a day before the presen-
tation. To collect the urine, the donor hamsters were
placed in metabolic cells with a mesh f loor, and the
MVGS was collected with a cotton swab and trans-
ferred to a plastic tube with 1 mL of distilled water.
To avoid the effect of the daily rhythms of hormone
secretion, the exposure of excretions and blood taking
were performed at the same time of day (from 10 to
11 a.m.). Filter paper with a diameter of 4 cm coated
with 100 μL of the donor excretion was placed into the
cell with the test animal. After a 30-minute exposure,
0.4–0.5 mL of blood was taken from each test animal
from the hyoid vein. This procedure took no more
than 30 s and did not cause a stress reaction in the ani-
mals, which could have led to the release of cortisol in
the blood (Graievskaya et al., 1986). The blood serum
was separated by centrifuging at 6000 rpm and stored
at a temperature of ‒18°C until the measurements
were taken. The results were processed in May–June
2016. To determine the concentrations of hormones
(testosterone, progesterone, and cortisol), heteroge-
neous enzyme-linked immunosorbent assays using an
iMark plate spectrophotometer (Bio-Rad) with com-
mercial sets of reagents by Immunotekh (Moscow,
Russia) were used.
The statistical assessment of the reliability of differ-
ences was carried out using the Wilcoxon matched-
pairs test and the Statistica 8.0 program.
1254 KROPOTKINA et al.

Table 1. Hormonal response of male Eversmann’s hamster to exposure to excretions of conspecific females, ng/mL
Exposure Cortisol Testosterone Progesterone
Fall
Control 84.58 ± 10.29 1.9 ± 0.50 0.13 ± 0.04
Urine 10.57 ± 2.94* 7.06 ± 2.10 0.35 ± 0.05*
MVGS 41.84 ± 22.53 5.21 ± 2.34 0.43 ± 0.14*
Winter
Control 49.17 ± 9.58 3.4 ± 1.10 0.16 ± 0.01
Urine 16.17 ± 5.19* 8.25 ± 2.46 0.23 ± 0.02*
MVGS 16.27 ± 4.14* 10.17 ± 2.85* 0.35 ± 0.08*
Spring
Control 49.32 ± 12.22 6.5 ± 1.40 0.11 ± 0.01
Urine 11.94 ± 2.89* 13.04 ± 2.18 0.27 ± 0.05*
MVGS 15.88± ± 2.72* 11.44 ± 1.35* 0.19 ± 0.01*
Summer
Control 18.73 ± 3.92 4.41 ± 0.63 0.11 ± 0.01
Urine 11.96 ± 1.46* 10.44 ± 2.59* 0.31 ± 0.05**
MVGS 12.65 ± 4.22 9.30 ± 2.25 0.19 ± 0.02
The reliability of differences from the control: * P < 0.05, ** P < 0.01.

RESULTS AND DISCUSSION winter period can be explained by better, compared

The nature of changes in testosterone, progester-
one, and cortisol in the blood of Eversmann’s ham-
sters in response to exposure to urine and MVGS of
conspecific females in different seasons of the year is
presented in Table 1.
In the fall period, no significant changes in the tes-
tosterone levels in the blood of males were recorded in
response to any of the signals presented. In winter and
spring, a reliable increase in testosterone (P < 0.05)
was caused only by the MVGS of females. In the sum-
mer, a similar response was recorded only for urine
exposure (P < 0.05). The level of cortisol decreased
significantly in response to the odor of female urine in
the summer and fall periods (P < 0.05); in winter and
in spring, a reliable decrease in the concentration of
cortisol was caused by both excretions investigated.
The presentation of urine and MVGS of conspecific
females significantly increased (P < 0.05) the proges-
terone levels in males in all seasons of the year, except
for the presentation of MVGS in the summer period.
Thus, the chemical signals capable of inducing a
reliable rise in testosterone levels in male Eversmann’s
hamsters were present both in urine and MVGS of
conspecific females, just as has been noted for a num-
ber of other species of the semifamily Cricetinae: the
Djungarian (Feoktistova, 2008; Feoktistova and Naid-
enko, 2006) and Chinese hamsters (Potashnikova and
Feoktistova, 2014).
The appearance of the chemical signals that sig-
nificantly increase the level of testosterone in the
blood plasma of males, in the MVGS of females in the
with urine, preservation of this secretion on the sub-
strate (owing to the fat basis). Consequently, the infor-
mation in it reaches the recipient individuals (i.e.,
males) more effectively, preparing them for encoun-
ters with females. Thus, already in winter, with an
increase in the length of the light day, males begin to
be interested in females (Kuznetsova et al., 2014).
In the summer period, only the smell of female urine
causes a significant increase in the testosterone level
(P < 0.05) in male conspecific animals. Thus, we can
suggest that the most active breeding in nature in the
investigated species occurs in the spring, as it is during
this season that all the males give the most pronounced
hormonal response to all the chemicals studied.
As was shown by the studies of V.S. Gromov and
V.V. Voznesenskaya (2013), the correlation of the tes-
tosterone levels, aggressiveness, and parental care
depend largely on the spatio-ethological structure of a
species. In some species the increased level of testos-
terone intensifies aggressive motivations and weakens
parental care, in others it can even stimulate the man-
ifestation of caring for the young on the part of the
father (Gromov, 2013; Gromov and Voznesenskaya,
2013). Progesterone in males can also stimulate paren-
tal care. For example, in male Campbell’s hamster of
the “western” phylogroup, the level of progesterone
before the birth of the brood and immediately after the
appearance of the young is higher than at other times
(Wynne-Edwards, 2003; Schum and Wynne-
Edwards, 2005).
Thus, a decrease in the level of cortisol (under the
influence of the smell of individuals of the opposite

BIOLOGY BULLETIN Vol. 44 No. 10 2017

 SEASONAL CHANGES IN THE HORMONAL RESPONSE
sex) in combination with a simultaneous increase in
the levels of testosterone and progesterone allows the
males of the species under study to achieve maximum
reproductive success, despite the high degree of intra-
specific aggression.
ACKNOWLEDGMENTS
This study was supported by the Russian Science
Foundation, project no. 16-14-10269.
REFERENCES
Bronson, F.H., Seasonal regulation of reproduction in
mammals, in Physiology Reproduction, Knobil, E. and
Neill, J., Eds., New York: Raven Press, 1988, vol. 2,
pp. 1831–1872.
Bronson, F.H., Mammalian Reproductive Biology, Chicago:
Univ. Chicago Press, 1989.
Buck, L.B., Olfactory receptors and odor coding in mam-
mals, Nutr. Rev., 2004, vol. 62, no. 11, pp. 184–188.
Feoktistova, N.Yu. and Naidenko, S.V., Hormonal response
to conspecific chemical signals as an indicator of seasonal
reproduction dynamics in the desert hamster, Phodopus rob-
orovskii, Russ J. Ecol., 2006, vol. 37, no. 6, pp. 426–430.
Feoktistova, N.Yu., Khomyachki roda Phodopus. Siste-
matika, filogeografiya, ekologiya, fiziologiya, povedenie, khi-
micheskaya kommunikatsiya (Hamsters of the Genus Pho-
dopus: Systematics, Phylogeography, Ecology, Physiology,
Behavior, and Chemical Communication), Moscow: Tov.
Nauch. Izd. KMK, 2008.
Ferkin, M.H., Sorokin, E.S., Renfroe, M.W., and John-
ston, R.E., Attractiveness of male odors to females varies
directly with plasma testosterone concentration in meadow
voles, Physiol. Behav., 1994, vol. 55, pp. 347–353.
Flint, V.E. and Golovkin, A.N., Essay on the comparative
ecology of hamsters of Tuva, Byul. MOIP. Otd. Biol., 1961,
vol. 66, no. 5, pp. 57–75.
Flint, V.E., Chugunov, Yu.D., and Smirin, V.M., Mlekopi-
tayushchie SSSR (Mammals of the USSR), Moscow: Mysl’,
1970.
Graievskaya, B.M., Surov, A.V., and Meshersky, I.G., The
tongue vein as a source of blood in the golden hamster,
Zeitschrift Versuchstierkunde, 1986, vol. 28, no. 1, pp. 41–
43.
Gromov, V.S., Zabota o potomstve u gryzunov: fiziologiches-
kie, etologicheskie i evolyutsionnye aspekty (Care of Young in
Rodents: Physiological, Ethological and Evolutionary
Aspects), Moscow: Tovar. Nauch. Izd. KMK, 2013.
Gromov, V.S. and Voznesenskaya, V.V., Care of young,
aggressiveness, and secretion of testosterone in male
rodents: a correlation analysis, Biol. Bull. (Moscow), 2013,
vol. 40, no. 5, pp. 463–470.
Jeyaraj, D.A., Maran, R.R.M., Aruldhas, M.M., and Gov-
indarajulu, P., Progesterone induced modulation of serum
hormonal profiles in adult male and female rats, Endocrine
Res., 2001, vol. 27, nos. 1–2, pp. 223–232.
Klevezal’, G.A., Feoktistova, N.Yu., Shchepotkin, D.V.,
and Surov, A.V., Features of recording hibernation on the
BIOLOGY BULLETIN Vol. 44 No. 10 2017
1255
surface of incisors of hamsters of the genus Allocricetulus,
Zool. Zh., 2015, vol. 94, no. 2, pp. 259–272.
Kotenkova, E.V., Comparative analysis of ethological and
physiological mechanisms of precopulative reproductive
isolation in rodents, Usp. Sovrem. Biol., 2014, vol. 134,
no. 5, pp. 488–518.
Kropotkina, M.V., Seasonal patterns of behavioral and hor-
monal responses of female Phodopus hamsters to the excreta
of conspecific males, in Aktual’nye problemy ekologii i evoly-
utsii v issledovaniyakh molodykh uchenykh (Actual Problems
of Ecology and Evolution in the Research of Young Scien-
tists), Moscow: Tovar. Nauch. Izd. KMK, 2012.
Kuznetsova, E.V., Kropotkina, M.V., Feoktistova, N.Yu.,
and Surov, A.V., Seasonal changes in body weight, levels of
sex steroids, and cortisol in male hamsters of the genus
Allocricetulus, Povolzh. Ekol. Zh., 2014, no. 4, pp. 529–536.
Leonard, S.T. and Ferkin, M.H., Prolactin and testosterone
mediate seasonal differences in male preference for the
odors of females and the attractiveness of male odors to
females, in Advances in Chemical Signals in Vertebrates,
Johnston, R.E., Muller-Schwarze, D., and Sorensen, P.W.,
Eds., New York: Kluwer Acad. Plenum Publ., 1999,
pp. 437–443.
Potashnikova, E.V. and Feoktistova, N.Yu., Seasonal char-
acteristics of hormone response of male Chinese hamsters
(Cricetulus b. griseus) to chemical signals of conspecific
females, Sens. Sist., 2014, vol. 28, no. 1, pp. 93–98.
Powers, J.B., Bergondy, M.L., and Matochik, J.A., Male
hamster sociosexual behaviors: effects of testosterone and its
metabolites, Physiol. Behav., 1985, vol. 35, no. 4, pp. 607–
616.
Rozhnov, V.V., Oposredovannaya khemokommunikatsiya v
sotsial’nom povedenii mlekopitayushchikh (Mediated Chemo-
communication in Social Behavior of Mammals), Moscow:
Tovar. Nauch. Izd. KMK, 2011.
Ryurikov, G.B. and Surov, A.V., On the causes of isolation
of ranges of gray and Eversmann hamsters in the Trans-
Volga region, in Bioresursy i bioraznobrazie ekosistem Povo-
lzh’ya: materialy mezhdunar. soveshch. (Bioresources and
Biodiversity of Ecosystems of the Volga Region: Proc. Int.
Conf.), Saratov: Izd. Sarat. Univ., 2005, pp. 181–185.
Ryurikov, G.B., Surov, A.V., and Tikhonov, I.A., Evers-
mann hamster (Allocricetulus eversmanni) in the Saratov
Trans-Volga region: ecology and behavior in nature, Povolzh.
Ekol. Zh., 2003, no. 3, pp. 251–258.
Sachs, B.D., Airborne aphrodisiac odor from estrous rats:
implication for pheromonal classification, in Advances in
Chemical Signals in Vertebrates, Johnston, R.E., Muller-
Schwarze, D., and Sorenson, P.W., Eds., New York: Kluwer
Acad. Plenum Publ., 1999, pp. 333–342.
Schum, J.E. and Wynne-Edwards, K.E., Estradiol and pro-
gesterone in parental and non-parental hamsters (Phodo-
pus) becoming father: conflict with hypothesized roles,
Hormones Behavior, 2005, no. 47, pp. 410–418.
Shchepot’ev, N.V., The winter activity of Eversmann ham-
ster, Priroda (Moscow, Russ. Fed.), 1959, no. 7, p. 113.
Shilov, I.A., Ekologo-fiziologicheskie osnovy populyatsion-
nykh otnoshenii u zhivotnykh (Ecological and Physiological
Bases of Population Relations in Animals), Moscow: Izd.
MGU, 1977.
1256 KROPOTKINA et al.
Shilov, I.A., Population homeostasis, Zool. Zh., 2002,
vol. 81, no. 9, pp. 1029–1047.
Sludskii, A.A., Bekenov, A., Borisenko, V.A., Grachev, Yu.A.,
Ismagilov, M.I., Kapitonov, V.I., Strautman, E.I., Fedo-
senko, A.K., and Shubin, I.G., Mlekopitayushchie Kaza-
khstana (Mammals of Kazakhstan), Alma-Ata: Nauka
KazSSR, 1977, vol. 1, part 2.
Tamar, G., Osnovy sensornoi fiziologii (Fundamentals of
Sensory Physiology), Moscow: Mir, 1976.
Ushakova, M.V., Feoktistova, N.Yu., Petrovskii, D.V.,
Gureeva, A.V., Naidenko, S.V., and Surov, A.V., Features
of hibernation of Eversmann hamster (Allocricetulus evers-
manni Brandt, 1859) from the Saratov Trans-Volga region,
Povolzh. Ekol. Zh., 2010, no. 4, pp. 415–422.
Vorontsov, N.N., Fauna SSSR. Mlekopitayushchie. Nizshie
khomyakoobraznye (Cricetidae) mirovoi fauny (Fauna of the
USSR. Mammals. Lower Cricetidae (Cricetidae) of the
World Fauna), Leningrad: Nauka. Leningr. otd., 1982.
Wilson, E.O., Chemical communication within animal
species, in Chemical Ecology, Sondheimer, E. and Sime-
one, J.B., Eds., New York: Academic Press, 1970, pp. 133–
155.
Wilson, J., The role of androgens in male gender role
behavior, Endocrine Rev., 1999, vol. 20, no. 5, pp. 726–737.
Wynne-Edwards, K.E., From dwarf hamster to daddy: the
intersection of ecology, evolution, and physiology that pro-
duces paternal behavior, Adv. Study Behav., 2003, vol. 32,
pp. 207–261.
Zucker, I., Johnston, P.G., and Frost, D., Comparative,
physiological and biochronometric analyses of rodent sea-
sonal reproductive cycles, Progr. Reprod. Biol., 1980, vol. 5,
pp. 102–103.
Translated by N. Smolina
BIOLOGY BULLETIN Vol. 44 No. 10 2017