Blocking oxytocin receptors inhibits vaginal marking to male odors in female Syrian hamsters

In Syrian hamsters (Mesocricetus auratus), precopulatory behaviors such as vaginal scent marking are essential for attracting a suitable mate. Vaginal marking is dependent on forebrain areas implicated in the neural regulation of reproductive behaviors in rodents, including the medial preoptic/anterior hypothalamus (MPOA-AH). Within MPOA-AH, the neuropeptide oxytocin (OT) acts to facilitate copulation (lordosis), as well as ultrasonic vocalizations towards males. It is not known, however, if OT in this area also facilitates vaginal marking. In the present study, a specific oxytocin receptor antagonist (OTA) was injected into MPOA-AH of intact female Syrian hamsters to determine if oxytocin receptor-dependent signaling is critical for the normal expression of vaginal marking elicited by male, female, and clean odors. OTA injections significantly inhibited vaginal marking in response to male odors compared with vehicle injections. There was no effect of OTA on marking in response to either female or clean odors. When injected into the bed nucleus of the stria terminalis (BNST), a nearby region to MPOA-AH, OTA was equally effective in decreasing marking. Finally, the effects of OTA appear to be specific to vaginal marking, as OTA injections in MPOA-AH or BNST did not alter general locomotor activity, flank marking, or social odor investigation. Considered together, these results suggest that OT in MPOA-AH and/or BNST normally facilitates male odor-induced vaginal marking, providing further evidence that OT generally supports prosocial interactions among conspecifics.     

The role of vaginal stimulation for the acquisition of conditioned place preference in female Syrian hamsters

This study investigated the importance of vaginal stimulation on conditioned place preference by sexual behavior in female Syrian hamsters. Hormonally primed female hamsters given vaginal masks and topical lidocaine prior to conditioning sessions were no different in their place preference conditioning compared to female controls with no vaginal mask or lidocaine. Control females not provided a sexually active male did not show any preference for either compartment of the conditioning apparatus. These results suggest that stimuli, in addition to vaginal stimulation, are effective in producing a place preference during the mating experience in female hamsters.     

Olfactory bulbectomy depresses ultrasound production and scent marking by female hamsters

Female hamsters were observed for rates of ultrasound production, vaginal marking and flank marking before and after bilateral olfactory bulbectomy, unilateral bulbectomy or sham operations. Unilaterally bulbectomized and sham-operated subjects maintained preoperative levels of each behavior. In contrast, females with bilateral olfactory bulbectomies showed consistent decreases in rates of vocalization and scent marking. These results emphasize the dependence on an intact olfactory system of several courtship or agonistic behaviors that affect hamster reproduction by functioning as noncontact communications. Together with the results of previous studies suggesting that lordosis by female hamsters is unaffected by olfactory bulbectomy, these data also suggest differences in the brain and sensory mechanisms that control distinct types of reproductive behavior.     

Paradigm for comparing sexual behavior and scent marking in male gerbils

Gerbils are popular subjects for research on social behavior and olfactory communication, yet little has been reported about their sexual behavior, apparently because researchers have found it difficult to elicit mating in stiuations where it can be observed and quantified. Here we describe a paradigm for eliciting complete copulatory behavior in male gerbils. We also report on the relationship between scent marking and sexual behavior and indicate how several environmental variables affect these behaviors. Our data indicate that (1) scent marking and sexual behavior are related, i.e., males that display sexual activity mark more often than males that do not mate; (2) male gerbils are more likely to copulate late in the light phase of the light: dark cycle than early in the dark phase; (3) housing males in pairs or groups facilitates copulatory behavior; and (4) marking frequencies as assessed in the sex arenas are positively correlated with, but higher than, marking scores obtained in the openfield.     

Mating preference and sexual reinforcement in female rats

Intact female rats chose between sexually active males and inactive males or females in a runway-choice apparatus. In either a discrete choice or continuous choice format, sexually receptive female rats demonstrated a preference for sexually active males over the other two incentive animals. No preference was demonstrated when females were sexually non-receptive. The results demonstrate that, for sexually receptive female rats, copulation can have reinforcing effects distinct from the reinforcing effects of more general social and exploratory factors.     

Lesions that functionally disconnect the anterior and posterodorsal sub-regions of the medial amygdala eliminate opposite-sex odor preference in male Syrian hamsters (Mesocricetus auratus)

In many rodent species, such as Syrian hamsters, reproductive behavior requires neural integration of chemosensory information and steroid hormone cues. The medial amygdala (MA) processes both of these signals through anatomically distinct sub-regions; the anterior region (MeA) receives substantial chemosensory input, but contains few steroid receptor-labeled neurons, whereas the posterodorsal region (MePD) receives less chemosensory input, but contains a dense population of steroid receptors. Importantly, these sub-regions have considerable reciprocal connections, and the goal of this experiment was therefore to determine whether interactions between MeA and MePD are required for male hamsters' preference to investigate female over male odors. To functionally disconnect MeA and MePD, males received unilateral lesions of MeA and MePD within opposite brain hemispheres. Control males received either unilateral lesions of MeA and MePD within the same hemisphere or sham surgery. Odor preferences were measured using a 3-choice apparatus, which simultaneously presented female, male and clean odor stimuli; all tests were done under conditions that either prevented or allowed contact with the odor sources. Under non-contact conditions, males with asymmetrical lesions investigated female and male odors equally, whereas males in both control groups preferred to investigate female odors. Under contact conditions, all groups investigated female odors longer than male odors, although males with asymmetrical lesions displayed decreased investigation of female odors compared to sham males. These data suggest that MeA–MePD interactions are critical for processing primarily the volatile components of social odors and highlight the importance of input from the main olfactory system (MOS) to these nuclei in the regulation of reproductive behavior. More broadly, these results support the role of the MA in integrating chemosensory and hormone information, a process that may underlie social odor processing in a variety of behavioral contexts.     

Estrogen and progesterone interaction in the regulation of scent marking in the female Mongolian gerbil (Meriones unguiculatus)

Ovariectomized female gerbils were treated with either 0, 20, 40, 80 or 160 μg estradiol benzoate (EB) followed in 36 hr by either 0, 250, 500 or 1000 μg progesterone. The animals were tested for ventral scent marking nine hr after progesterone treatment. Treatment with 20, 40 or 80 μg EB resulted in a significant increase in marking in the ovariectomized female for each dose of progesterone used. One hundred and sixty μg EB was not effective while 80 μg was the most effective dose at each dose of progesterone. There were no differences among the 80 μg EB groups in mean marking frequency as long as progesterone was present. Neither estrogen alone nor progesterone alone was effective in stimulating a significant level of marking. An additional sample of ovariectomized females were maintained for four weeks on either 2.5, 5, 10, 20 or 40 μg EB and tested for marking each week 45 hr after EB administration. On the fifth week 250 μg progesterone was given each week 36 hr following EB with behavioral testing nine hr later. Such a regime was followed for four weeks. Marking remained at zero levels on weeks one through four. EB + progesterone on weeks five through eight, however, resulted in a significant increase in marking in animals receiving 10 μg EB (mean = 9.0 ± 3.5), 20 μg EB (mean = 6.6 ± 2.5) and 40 μg EB (mean = 9.2 ± 4.2). It was concluded that both estrogen and progesterone were essential for the support of marking in this sample of low-marking females and that since there were no reliable differences between groups receiving different doses of progesterone, estrogen was the primary stimulus controlling marking behavior, with progesterone perhaps playing a modifying role.     

Regulation of scent marking in the female Mongolian gerbil Meriones unguiculatus

The role of estrogens and progesterones in the control of scent marking in the female Mongolian gerbil (Meriones unguiculatus) has remained unclear. Two experiments were conducted which attempted to assess the ovarian contribution to the regulation of this behavioral pattern. In Experiment 1, ten groups of ovariectomized females were treated with one or a combination of steroid hormones which had been selected on the basis of being known secretory products of the ovaries of other rodent species. Only testosterone propionate or a combination of testosterone propionate and estrogen were effective in stimulating marking. Both androgens and estrogens supported gland growth whereas only estrogens supported uterus growth. In a second experiment, androstenedione, a primary precursor to testosterone and estrogen, was administered to intact and ovariectomized females for a period of four weeks. Each female received a weekly marking test following treatment. Intact females responded with a rapid and significant increase in marking frequency whereas ovariectomized and intact control females continued to mark at control levels. It was concluded that the conversion of androstenedione or related hormones into behaviorally active compounds during period of high marking (e.g., as during late gestation and lactation and lactation) is at least one means by which marking is regulated in the female gerbil.     

Re-feeding and the restoration of odor attractivity, odor preference, and sexual receptivity in food-deprived female meadow voles

Food-deprived meadow voles were used to test predictions of two hypotheses associated with the recovery of sexual behaviors following re-feeding. Specifically, we tested between the body weight set point and metabolic fuels hypotheses. To do so, we determined whether the body weight of previously food-deprived female voles had to return to pre-food deprivation levels before they would recover their sexual behaviors. The body weight set point hypothesis predicts that food-deprived females that were re-fed will recover their sexual behavior after they return to their original body weight. In contrast, the metabolic fuels hypothesis predicts that food-deprived females that were re-fed will recover their sexual behavior before they return to their original body weight. That is, when the females are in positive energy balance. To distinguish between these two hypotheses, female voles were food deprived for 24 h, which is sufficient to inhibit all three components of sexual behavior. The food-deprived females were then supplied ad libitum food for 0 h, 24 h, 48 h, 72 h, or 96 h and weighed. Females were then tested for their sexual behaviors (odor attractivity, odor preference or proceptivity, sexual receptivity). Re-feeding for 48 h was sufficient to restore odor attractivity, 72 h was sufficient to restore odor preferences for opposite sex conspecific odors, and 96 h was sufficient to restore sexual receptivity to those similar to that of females that were not food deprived. The time-points that the behaviors were recovered were prior to voles recovering their initial body weight. Thus, the data support the metabolic fuels hypothesis.     

X-ray kinematics analysis of vaginal scent marking in female Syrian hamsters (Mesocricetus auratus)

Vaginal marking is a stereotyped scent marking behavior in female Syrian hamsters used to attract male hamsters for mating. Although the modulation of vaginal marking by hormones and odors is well understood, the motor control of this proceptive reproductive behavior remains unknown. Therefore, we used X-ray videography to visualize individual bone movements during vaginal marking. Kinematic analyses revealed several consistent motor patterns of vaginal marking. Despite exhibiting a diversity of trial-to-trial non-marking behaviors (e.g. locomotor stepping), we found that lowering and raising the pelvis consistently corresponded with coordinated flexion and extension cycles of the hip, knee, and tail, suggesting that these movements are fundamental to vaginal marking behavior. Surprisingly, we observed only small changes in the angles of the pelvic and sacral regions, suggesting previous reports of pelvic rotation during vaginal marking may need to be reconsidered. From these kinematic data, we inferred that vaginal marking is primarily due to the actions of hip and knee extensor muscles of the trailing leg working against gravity to support the weight of the animal as it controls the descent of the pelvis to the ground. The cutaneous trunci muscle likely mediates the characteristic flexion of the tail. Interestingly, this tail movement occurred on the same time scale as the joint kinematics suggesting possible synergistic recruitment of these muscle groups. These data therefore provide new targets for future studies examining the peripheral control of female reproductive behaviors.     

Recognition of competitors by male golden hamsters

Golden hamsters, like many animals, form dominant/subordinate relationships after aggressive encounters. We examined whether behavioral responses by males that won or lost fights would differ toward familiar and unfamiliar male stimulus animals. In Experiment 1, male winners or losers of fights explored an arena containing a confined stimulus animal that was either familiar or novel and had either won or lost a fight. Compared to dominant males, losers spent less time in proximity to stimulus males and investigated them less. Losers also displayed higher levels of stretch-attend postures (indicative of risk assessment) than winners, and they showed more escape and locomotion in response to familiar winners than to unfamiliar winners, indicating recognition of the male that they had lost to. In Experiment 2, losers scent marked less to the odors of a familiar winner than to those of an unfamiliar winner. Thus, male hamsters appear to use familiarity with a former opponent's odors to adaptively regulate their responses to variations in social threat.     

Partner preference: Assessing the role of the female goat

For domestic ruminants, the male is thought to have the control over mate choice. The aim of this study was to assess the potential role the female goat plays in choosing a sexual partner. Approach frequency and proximity time to various social or sexual stimuli in a Y-maze apparatus were used as partner preference assessment parameters. Ovariectomized females were either induced into estrus (n = 8) or induced into anestrus (n = 8) and exposed to various goal stimuli — intact unfamiliar females (estrous and diestrous), intact males, and neutral (empty) in six separate trials. Total arm entries and time spent within one body length of the goal (proximity time — used to calculate preference scores) were recorded for subjects. With a male present, anestrous subjects had significantly more total arm entries than estrous subjects. Estrous subjects showed a significant preference for the male goal (M) versus the estrous female (ES), diestrous female (DS), and empty goals, respectively. Estrous subjects also showed a preference for the ES and DS goals when one goal was left empty. Anestrous subjects showed a preference for the goal containing ES, DS, and M compared to empty goals. They also showed a preference for ES vs. DS. The Y-maze is a tool that can be used to assess sexual and social partner preferences in female goats. The role the female plays in choosing a partner may be much greater than previously considered.     

The effects of neonatal androgenization of female hamsters on adult preference for female hamster vaginal discharge

The effects of neonatal exposure of hamsters to androgen on the development of an adult preference for female hamster vaginal discharge (FHVD) were investigated. Three groups treated on Days 2-3 postpartum (females administered testosterone propionate (TP) in a peanut oil base, females administered oil alone, and males administered oil alone) were combined factorially with four sequential adult conditions (intact, post-gonadectomy, post-gonadectomy plus TP therapy, and post-gonadectomy) to produce twelve experimental conditions. Statistical analysis indicated that neonatal oil females did not show a significant FHVD preference in any adult condition. Neonatal TP females displayed a significant FHVD preference only in the post-gonadectomy plus TP condition. Males retained a significant FHVD preference in all the adult conditions, although the preference shown in the two adult hormonal conditions (intact and post-gonadectomy plus TP) was significantly higher than the preference shown in the two nonhormonal conditions.     

X-ray kinematics analysis of vaginal scent marking in female Syrian hamsters (Mesocricetus auratus)

Vaginal marking is a stereotyped scent marking behavior in female Syrian hamsters used to attract male hamsters for mating. Although the modulation of vaginal marking by hormones and odors is well understood, the motor control of this proceptive reproductive behavior remains unknown. Therefore, we used X-ray videography to visualize individual bone movements during vaginal marking. Kinematic analyses revealed several consistent motor patterns of vaginal marking. Despite exhibiting a diversity of trial-to-trial non-marking behaviors (e.g. locomotor stepping), we found that lowering and raising the pelvis consistently corresponded with coordinated flexion and extension cycles of the hip, knee, and tail, suggesting that these movements are fundamental to vaginal marking behavior. Surprisingly, we observed only small changes in the angles of the pelvic and sacral regions, suggesting previous reports of pelvic rotation during vaginal marking may need to be reconsidered. From these kinematic data, we inferred that vaginal marking is primarily due to the actions of hip and knee extensor muscles of the trailing leg working against gravity to support the weight of the animal as it controls the descent of the pelvis to the ground. The cutaneous trunci muscle likely mediates the characteristic flexion of the tail. Interestingly, this tail movement occurred on the same time scale as the joint kinematics suggesting possible synergistic recruitment of these muscle groups. These data therefore provide new targets for future studies examining the peripheral control of female reproductive behaviors.     

Effects of intracerebral implants of steroid hormones on scent marking in the ovariectomized female gerbil (Meriones unguiculatus)

The effects of intracerebral implants of steroid hormones on scent marking in the female gerbil (Meriones unguiculatus) were studied in two experiments. In Experiment 1 various steroids were implanted alone or in combination into the preoptic and anterior hypothalamic area of ovariectomized females. Unilateral implants of testosterone + estrogen, estrogen, estrogen + progesterone, testosterone and testosterone + progesterone stimulated a significant level of marking when compared to controls. Experiment 2 utilized bilateral implants of estrogen dissolved in paraffin in order to explore the distribution hormone sensitive areas in the brain which might be important in the regulation of scent marking in the female gerbil. Pellets of estrogen-paraffin were implanted stereotaxically into either the anterior hypothalamus, preoptic area, septum, hippocampus, thalamus, amygdala or anterior olfactory nucleus of ovariectomized females. Total dosage of hormone implanted was 8.2–8.4 μg. A significant level of marking resulted in animals receiving implants into the anterior hypothalamus, preoptic area and septum when compared to controls. Marking appeared at about the same rate in each of these groups; however, the level of marking attained differed. By the last trial, anterior hypothalamic implanted animals were marking significantly more often than animals in either the preoptic or septum groups. Although there was no evidence of ieakage from the brain, the data suggested that some rapid diffusion of hormone, largely restricted to the brain, was taking place or that the three areas were differentially responsive to the hormone. The data do indicate that some localization of function does exist with respect to regulation of scent marking in the female.     

Scent marking and mate choice in the golden hamster

Test groups consisting of three males and one female were adapted to home areas within a 244 X 122 cm enclosure for a 4-day period. Subsequently, agonistic interactions and patterns of scent marking were recorded daily for 30 min over the duration of the female's 4-day estrous cycle. Alpha males flank marked significantly more than the two subordinates. The former accounted for over 95% of flank gland marking by males on the two days prior to the female's period of sexual receptivity. The female showed increased frequencies of both flank gland and vaginal marking over the three nonreceptive days of the cycle with the highest frequency of marking occurring 24 hr prior to sexual receptivity. During the two days prior to receptivity, the female both vaginal marked and flank marked significantly more in and about the home area of the alpha male. We conclude that the female hamster shows mate choice prior to the onset of sexual receptivity and that both vaginal marks and flank marks are employed to facilitate mating with the alpha male.     

The role of the medial preoptic area in appetitive and consummatory reproductive behaviors depends on sexual experience and odor volatility in male Syrian hamsters

In Syrian hamsters (Mesocricetus auratus), the expression of reproductive behavior requires the perception and discrimination of sexual odors. The behavioral response to these odors is mediated by a network of ventral forebrain nuclei, including the medial preoptic area (MPOA). The role of MPOA in male copulatory behavior has been well-studied, but less is known about the role of MPOA in appetitive aspects of male reproductive behavior. Furthermore, many previous studies that examined the role of MPOA in reproductive behavior have used large lesions that damaged other nuclei near MPOA or fibers of passage within MPOA, making it difficult to attribute post-lesion deficits in reproductive behavior to MPOA specifically. Thus, the current study used discrete, excitotoxic lesions of MPOA to test the role of this nucleus in opposite-sex odor preference and copulatory behavior in both sexually-naïve and sexually-experienced males. Lesions of MPOA eliminated preference for volatile, opposite-sex odors in sexually-naïve, but not sexually-experienced, males. When males were allowed to contact the sexual odors, however, preference for female odors remained intact. Surprisingly, lesions of MPOA caused severe copulatory deficits only in sexually-naïve males, suggesting previous reports of copulatory deficits following MPOA lesions in sexually-experienced males were not due to damage to MPOA itself. Together, these results demonstrate that the role of MPOA in appetitive and consummatory aspects of reproductive behavior varies with the volatility of the sexual odors and the sexual experience of the male.     

Effects of vomeronasal organ removal on individual odor discrimination, sex-odor preference, and scent marking by female hamsters

Removal of the vomeronasal organ (VNX) did not eliminate the ability of female hamsters to discriminate between individual male's flank gland or urine odors in a habituation/discrimination task nor did it impair preference for male odors over female odors from a distance. Vomeronasal organ removal did reduce overall levels of investigation of flank gland odor in the habituation/discrimination task. Although VNX females did not show severe impairments in the frequency of either flank or vaginal marking in response to odors, they did show an abnormal pattern of marking. VNX females, unlike shams, did not flank mark more to female odors than to male odors, nor did they vaginal mark more to male odors than to female odors. Thus, the vomeronasal organ in female hamsters appears to be important for differences in scent marking toward male and female odors, but is not essential for discrimination of individual odors or for preferences for male over female odors.     

Juvenile play conditions sexual partner preference in adult female rats

Rats can display a conditioned partner preference for individuals that bear an odor previously associated with sexual reward. Herein we tested the possibility that odors associated with the reward induced by social play in prepubescent rats would induce a conditioned partner preference in adulthood. Two groups of 31-day-old, single-housed female rats were formed, and were given daily 30-min periods of social play with scented females. In one group, almond scent was paired with juvenile play during conditioning trials, whereas lemon scent functioned as a novel odor in the final test. The counterbalanced group received the opposite association. At age 42, females were tested for play partner preference with two males, one almond-scented and one lemon-scented. In both groups females displayed a play partner preference only for males scented with the paired odor. They were ovariectomized, hormone-primed, and at age 55 were tested for sexual partner preference with two scented stud males. Females displayed a sexual preference towards males scented with the paired odor as observed with more visits, solicitations, hops and darts, intromissions and ejaculations. These results indicate that olfactory stimuli paired with juvenile play affects later partner choice for play as well as for sex in female rats.     

A competitive effect of androgen signaling on male mouse attraction to volatile female mouse odors

Olfaction plays an important role in animal communication. We hypothesized that males recognize the attractive volatile odors attributed to female reproductive ability. We measured the period during which a male mouse spent sniffing volatile odors from a sham-operated female mouse or an ovariectomized mouse without visual or tactile contact. Intact male mice spent more time sniffing volatile odors from proestrous, estrous or metestrous females than from ovariectomized females. There was no difference in castrated male mice. To investigate the involvement of sexual hormone in this behavior, castrated male mice were treated with 17 alpha-estradiol (E), dihydrotestosterone (DHT), or both. E-treatment did not affect sniffing behavior. Regardless of the estrous stages, DHT-treated castrated males spent less time sniffing the volatile odors from sham-operated than from ovariectomized female mice. Both E- and DHT-treated castrated males spent less time sniffing the volatile odors from proestrous or estrous females than from ovariectomized females. These results suggest that neither androgen nor estrogen is sufficient for reproducing male attraction to volatile female mouse odors, and that androgen signaling has a competitive effect against the attraction.