Effects of ventromedial nucleus lesions on the display of lordosis behavior in the male rat. Interactions with facilitory effects of male urine

Previous observations showed that exposure to the odor of male urine prior to mating could enhance the display of lordosis behavior in male rats feminized with ovarian hormones. This study was performed to determine in feminized male rats whether the control of lordosis behavior by the olfactory system was mediated by the ventromedial nucleus (VMN) of the hypothalamus. Male rats were orchidectomized (ORCH) as adults and primed with 25 micrograms estradiol benzoate (EB) and 150 micrograms progesterone (P) 40 hr apart. Lordosis behavior was tested 9 +/- 1 hr after P injection. VMN lesions were shown to completely suppress the display of lordosis behavior as compared to sham VMN operated and dorsomedial nucleus (DMN) lesioned animals. Exposure of feminized rats to the odor of male urine by 9 +/- 1 hr before mating significantly increased the proportion of ORCH rats that displayed lordosis behavior in response to male mounts. This effect was abolished by VMN lesions but was maintained in the sham VMN operated and DMN lesioned animals. These results were discussed in the light of the present knowledge on the neuroendocrine and olfactory structures which mediate lordosis behavior in the male rat.     

Hormonal control of the perception of the olfactory signals which facilitate lordosis behavior in the male rat

This study was designed to evaluate in the male rat the hormonal requirements for the facilitation of feminine behavior by the odor of male urine. Wistar rats from the WI and WII strains in our colony were orchidectomized (ORCH) as adults. A first group was given a single dose of 75 micrograms estradiol benzoate (EB) and tested for lordosis behavior 48 hr later. Exposure to the odor of male urine by 9 +/- 1 hr before the behavioral session did not increase the number of animals showing lordosis behavior as compared to non exposed controls. A second group of WI rats was given 0.5 micrograms EB every day for 4 to 8 days. A similar number of animals displayed lordosis behavior irrespective of whether they were exposed to the odor of urine before testing. A third group of WI rats was injected with 75 micrograms EB and 1 mg progesterone (P) 39 hr apart. Exposure to the odor of urine during estrogen treatment remained ineffective but significantly increased the number of animals showing lordosis behavior when performed at the time of P injection. A last group of WII rats was given 25 micrograms EB and 100 micrograms or 150 micrograms P 39 hr apart. Although uncapable as such to facilitate lordosis behavior the dose of 100 micrograms P rendered the animals responsive to the odor of urine. It was concluded that (1) the perception by feminized males of olfactory signals from the male was dependent on P; (2) an interaction between hormonal and sensory mechanisms was involved in the facilitation of lordosis behavior in the male rat.     

Sexual and olfactory preference in noncopulating male rats

In some species including rats, mice, gerbils, and rams, apparently normal males fail to copulate when repeatedly tested with receptive females. These animals are called "noncopulators (NC)," and the cause of this behavioral deficit is unknown. It has been shown that NC rats do not have hormonal alterations or deficits in the mechanisms that control penile function. The present study was designed to examine (Experiment 1) whether NC male rats prefer receptive females to sexually active males. In addition, the olfactory preference for bedding soiled from estrous or for anestrous bedding was investigated. These tests were performed in NC and copulating (C) male rats when the subjects were intact, gonadectomized (GDX), or GDX and treated with high doses of testosterone propionate (TP). Our results demonstrate that NC rats do not display sexual behavior even after high TP treatment. While C male rats have a clear preference for receptive females as opposed to a sexually active male, NC rats do not. In all hormonal conditions, the preference shown by NC rats for estrous bedding was significantly reduced in comparison to that seen in C rats. TP treatment in NC rats did not modify either partner or odor preference. In Experiment 2, we evaluated if NC rats are feminized and if it could be easier to induce feminine-like behavior by hormone treatment with estradiol benzoate (EB) or with EB plus progesterone (P) (EB+P). Odor preference for estrous or male bedding under these hormonal conditions was also compared. No differences between NC and C rats were found in feminine sexual behavior. In the olfactory test, we found that NC rats prefer odors from receptive females as opposed to male odors, but this preference is reduced compared to that of C rats. Males treated with EB or EB+P show no preference for female odors. These results demonstrate that treatment with EB or EB+P does not increase feminine sexual behavior in NC rats.     

LiCl aversive conditioning has transitory effects on pheromonal responsiveness in male house mice (Mus domesticus).

Appetitive and aversive experiences influence whether odors elicit precopulatory behavior from male rodents. A role for aversive experience in odor-elicited reproductive behaviors had been demonstrated for hamsters and rats, but similar work on house mice had not been performed. Four experiments examined whether lithium chloride (LiCl) aversive conditioning would alter two precopulatory behaviors (ultrasonic vocalizations and olfactory preference) that male house mice normally exhibit to female urine. Lithium chloride was used to aversively condition male house mice to either female urine odor, female urine in drinking water, the female herself, or a novel odor. Independent tests of taste aversion establishment were also conducted. In these experiments, LiCl aversive conditioning produced robust taste aversions to water adulterated with either female urine or a novel odorant/tastant (isoamylacetate), but only transitory decrements in odor-elicited, male-typical precopulatory behaviors. We conclude that aversive conditioning is unlikely to be a significant factor affecting male mouse precopulatory behavior.     

Enhanced synaptic responses in the piriform cortex associated with sexual stimulation in the male rat

Male rats that copulate to ejaculation with female rats bearing an odor show a learned preference to ejaculate selectively with females that bear the odor. This conditioned ejaculatory preference reflects an association between the odor and the reward state induced by ejaculation. Although little is known about the neuronal mechanisms that mediate this form of learning, convergence of genitosensory and olfactory inputs occurs in both hypothalamic and cortical regions, notably within primary olfactory (piriform) cortex, which may be involved in the encoding or storage of the association. The present study contrasted the ability of genital investigations, mounts, intromissions, ejaculations, and a sexually conditioned olfactory stimulus, to enhance evoked synaptic field potentials in the piriform cortex. Rats in the Paired group underwent conditioning trials in which they copulated with sexually receptive females bearing an almond odor. Rats in the Unpaired control group copulated with receptive females bearing no odor. Responses in the piriform cortex evoked by electrical stimulation of the olfactory bulb were recorded in male rats as they engaged in different aspects of sexual behavior, and were also recorded after conditioning, during exposure to cotton swabs bearing the almond odor. The monosynaptic component of responses was increased during intromission and ejaculation, and the late component of responses was increased during anogenital sniffing and mounting (with or without intromission). However, no differences in the amplitudes of evoked responses were found between the Paired and Unpaired groups, and no differences in synaptic responses were found during presentation of the odor after conditioning. These data indicate that short-term alterations in synaptic responsiveness occur in piriform cortex as a function of sexual stimulation in the male rat, but that responses are not significantly altered by a conditioned odor.     

The critical role of familiar urine odor in diminishing territorial aggression toward a castrated intruder in mice

Sensory chemo-signals conveying information on sex and familiarity are important to the manifestation of aggressive behaviors in male mice. In this study, we examined the role of familiarity conveyed by urine odor in the induction of aggressive behavior using a resident-intruder paradigm. First, an intact ICR male mouse (resident) was grouped with a castrated DBA mouse (cage-mate) and a female ICR mouse to allow the resident mouse to establish its territory. The resident male showed vigorous aggression, not only toward intact male DBA intruders, but also toward unfamiliar castrated DBA mice (UFC). In contrast, the aggression was markedly reduced toward its castrated DBA cage-mate. Next, to reveal how residents discriminate their cage-mates from unfamiliar intruders, we examined whether urine odor affected this familiarity-related aggression. When part of the body surface of a UFC was swabbed with the urine of a resident's cage-mate, the resident attacked the UFC much less often. These results suggest that the information about familiarity conveyed by urine odor plays an important role in controlling the territorial aggression of a resident male mouse toward castrated intruders.     

Urinary testosterone levels in the male blind mole rat (Spalax ehrenbergi) affect female preference

This study investigated the sexual attraction of female blind mole rats to four groups of male mole rats: (a) intact males raised in captivity; (b) intact males trapped in the field; (c) captive males injected with testosterone; (d) captive castrated males. In the first part we measured blood testosterone, androstenedione, and dihydrotestosterone (DHT) levels, by radioimmunoassay; and urine testosterone levels, measured by GC-MS. The second part examined the relationship between urine testosterone levels in males and their attractiveness to females. Higher blood and urine testosterone levels were found in the field animals and in those injected with testosterone compared to captive intact or castrated animals: urine testosterone levels in the two other groups were not detectable. Blood androstenedione levels were also higher in the field animals and in those injected with testosterone compared to captive intact or castrated mole rats. Blood dihydrotestosterone levels were not detectable in all four experimental groups. Female mole rats chose to spend a longer period of time next to males with high blood and urine testosterone levels and high blood androstenedione levels than next to those with lower levels of these hormones. Because courtship and sexual behavior are influenced both by high levels of blood and urine testosterone and high levels of blood androstenedione, we suggest that the low levels of courtship and other sexual behavior in captive mole rats may be related to the lack of female attraction to these males, which display low levels of all three parameters.     

Corticosterone increases depression-like behavior, with some effects on predator odor-induced defensive behavior, in male and female rats

This experiment examined the effect of repeated corticosterone injections on anxiety and depression-like behavior in male and female rats. Rats received either corticosterone or vehicle injections for 21 consecutive days prior to behavioral testing in the forced swim, open-field, and predator odor tests. The corticosterone injections significantly increased depression-like behavior in the forced swim test in both male and female rats but had no significant effect on anxiety in the open-field test. In the predator odor test, the corticosterone injections significantly increased a subset of defensive behaviors in the male rats. These results suggest that repeated exposure to corticosterone increases depression-like behavior, with some effects on anxiety, and that male rats may be more affected than female rats by this manipulation.     

Ovarian hormones and their role in aggression inhibition among male mice

The role of ovarian steroids in the inhibition of isolation-induced aggression (defined here as fighting or threatening behavior) in male mice was studied, as well as the possibility of interspecies action of mammalian pheromones. Male mice were paired and tested for aggressiveness after being smeared with urine from female mice, rats, rabbits, and humans. Urine from intact and sexually mature animals caused a decrement in aggression, while ovariectomized or sexually immature animals did not produce an aggression inhibiting urine. Urine from ovariectomized animals given injections of estradiol (0.1 mg/day) for 5 days, and other estrogens caused non-aggression. Heavy perfume did not stop aggression, supporting the theory that non-aggression is not the result of simple masking of male odors. Progesterone (1 mg/day for 5 days) was ineffective in inhibiting aggression. An interspecies action of this aggression inhibiting pheromone appears to exist among mice, rats, rabbits, and to some degree, humans.     

Androgen effects on responsiveness to aggression and stress-related odors of male mice

The effect of male gonadal hormones on responsiveness to the aggression and stress-related odors of male mice was examined. The initial experiment indicated that intact male mice would avoid an area of an open field that had been spotted with the urine of aggressive male donors, while castration of the subjects eliminated the response. Hormone replacement was effective in reinstating the aversion, clearly demonstrating the androgen-dependent nature of the response. A second experiment determined that the effects of castration do not generalize to another type of aversive odor, namely the alarm odors of castrate donors. That is, both intact and castrate males exhibited a pronounced aversion to the urine odors of castrates that had been subjected to a prolonged period of stress. These results suggest that gonadal hormone effects on olfactory responsivity are somewhat specific, and more interestingly, that the mechanism behind the effects of gonadal hormones on rodent aggression may lie in their influence on the nature of the response to the relevant olfactory stimuli.     

Development of sex differences in the response of spiny mouse pups to adult male odors

Using a three-choice preference test, olfactory-mediated investigatory activity in response to adult male urine odor was examined in a precocially active rodent, the spiny mouse (Acomys cahirinus) aged between 3-26 days. Temporally related sex differences were seen in the time spent in the presence of the odors of father's or unfamiliar adult male's urine, or distilled (control) water. Neither male nor female pups discriminated between odors from the father and strange adult males. After the first olfactory test, when the pups were aged between four and six days, male pups strongly preferred to stay in the vicinity of urine odors of adult males, whereas female pups avoided odors of adult males and remained in the enclosure with the control odor source. To our knowledge this is the first time that such a behavioral sex difference related to olfaction has been shown to occur in young rodent pups. We suggest that the sexually dimorphic response of the pups is associated with the development of later sex differences in behavior.     

Attraction of male guinea pigs to conspecific urine

The attraction of sexually experienced male guinea pigs to urine from a variety of sources was tested. Using a two-choice procedure, males were shown to be more attracted to urine from intact, nonreceptive females, urine from adult males castrated within 2 days of birth and urine from ovariectomized females, than to urine from intact adult male guinea pigs. Urine from intact males was preferred to urine from nonreceptive female Galea musteloides. Urine from males castrated as adults was not discriminated from urine from sham operated controls when collected 4 days postoperatively. However, castrate urine collected 14 days or greater postoperatively was preferred to the control urine. Urine from males and females aged 8–10 days was equally attractive but by age 18–20 days female urine was preferred to male urine. Further tests indicated female urine is preferred over male urine when only olfactory cues are available and that mixtures of male urine and female urine are preferred to male urine alone or diluted with water.     

Ultrasonic vocalizations and aggressive behavior in male rats

The aim of the present study was to know whether 22-28-kHz vocalizations have any communicatory role in the regulation of aggressive behavior in male rats of the Wistar strain. In pairs of intact rats 22-28-kHz vocalizations showed a positive correlation with the extent of aggressive behavior. The pattern of aggressive behavior during ultrasonic vocalizations was different from that just before and just after the vocalizations. However, surgically deafened rats were less active in aggressive behavior and more active in ambulatory activity in the open field than the controls. Muted rats were not different from the controls in both aggressive behavior and ambulatory activity. The present result that the deprivation of ultrasonic signals failed to increase aggressive behavior does not support the classical hypothesis that ultrasonic vocalizations inhibit the initiation of aggressive behavior. It is concluded that ultrasounds emitted during aggressive encounters may have little communicative value in male rats.     

Urinary odors and size protect juvenile laboratory mice from adult male attack

For a portion of their ontogeny, the juveniles of many mammalian species appear resistant to aggressive attack by adult male conspecifics. The possibility that urinary odors and small size contribute to this immunity from attack was investigated with C57/Bl, A/J, and DW/J laboratory mice. The results of the 1st of 6 experiments were that juvenile mice of both genders evoked great curiosity from aggressive resident male adults, but juveniles were rarely attacked. In Experiments II and III, exchanging urine between juveniles and adult males suggested that juveniles possess a distinctive odor. In Experiments IV–VI, dwarf adults, normal-sized adults, and juveniles were placed with aggressive adult male residents. The dwarf males were attacked, but not as severely as normal-sized male intruders (Experiment IV). The dwarf males, however, evoked more aggressive behavior than juveniles (Experiment V), and juveniles with the odors from either normal-sized or dwarf adult males were attacked more readily than non-odorized juveniles (Experiment VI). These data suggest that both odor and size contribute to the juveniles' immunity from attack.     

Sexual impairment of inexperienced male rats following pre- and postpuberal olfactory bulbectomy.

Olfactory bulbectomy of sexually inexperienced male rats prevented the occurrence of sexual behavior in most of the operated animals. This effect was observed in rats bulbectomized before puberty as well as after puberty. Bulbectomized rats which had heterosexual experience prior to the operation did not deviate from intact rats. Males living in cohabition with intact males prior to the operation showed only minor deficits in their mating performances. It was concluded that the olfactory lobe while of only minor importance for maintenance of mating once sexual behavior has been initiated, plays an important role in initiation of sexual behavior of the male rat. Furthermore, since treatment with testosterone of prepuberally bulbectomized rats did not stimulate the animals to sexual activity it was concluded that the sexual impairment following bulbectomy of isolated males presumably is not due to an impaired production of gonadal secretions.     

Morphine treatment and reproductive condition alter olfactory preferences for pup and adult male odors in female rats

Administration of morphine sulfate (MS) to pregnancy-terminated and postpartum lactating female rats disrupts both maternal behavior and postpartum aggression. Since the display of these behaviors may be heavily dependent on olfactory cues provided by the stimulus animals (rat pups and adult male rats, respectively), we examined whether MS was affecting the perception of the olfactory stimuli, and whether olfactory perception was modified by reproductive condition. In Experiment 1, lactating rats had their pups removed and were injected with MS (5.0 mg/kg, sc.) or saline. 60 min later they were placed into a two-choice apparatus, one side of which contained bedding soiled by neonates and the other clean bedding. Time spent on each side was recorded for a total of 5 min (300 s; chance = 150 s). Saline-treated mothers spent significantly more time on the pup-odor side, whereas MS-treated females spent significantly less. In Experiment 2, lactating females were treated with MS or saline and exposed to male odors (soiled bedding). MS significantly increased time spent on the side with male odors; when treated with saline, time spent was significantly reduced. Thus, in lactating rats. MS creates an aversion for pup odors while reducing the female's normal aversion toward male odors. In Experiment 3, ovariectomized (ovx) virgin females expressed neither an aversion nor a preference for the odor of pups following saline administration. After MS treatment, however, the virgins showed a distinct preference for pup odors. When exposed to male odors in Experiment 4 ovx virgins showed a marked preference for male odors after MS treatment, and neither a preference nor an aversion after saline. Experiment 5 examined pup odor preferences in intact virgins, early (Day 7), middle (Day 14), late-pregnant (Day 21), and prepartum (Day 22) rats. The pup odor preferences of virgin, Day 7, Day 14, and Day 21 pregnant rats were not different and generally were at chance levels. Day 22 pregnant females exhibited a marked preference for pup odors compared to chance levels, as well as compared to the other four groups. These findings suggest that opiates and endogenous opioids may regulate olfactory preferences and that alterations in this system may underlie normal behavioral changes toward conspecifics prepartum as well as during lactation.     

Sexually dimorphic enhancement by estradiol of male urinary odor detection thresholds in mice

We asked whether sex and adult estrogen exposure influence the detection thresholds for urinary odors used by mice to guide their social behaviors. Gonadectomized (GDX) male and female mice were trained on a two-choice food-motivated task to determine detection thresholds for male urinary odors. There was no significant sex difference in the detection of these odors by GDX subjects without hormone replacement. However, during treatment with estradiol benzoate (EB), GDX females, but not GDX males, showed an enhanced ability to detect these odors. To investigate a possible mechanism for this effect, the authors measured GDX females' odor-sampling behavior (sniffing) by monitoring intranasal pressure transients during performance of the urinary odor detection task with and without EB treatment. Under both hormone conditions, females decreased their sniffing frequency as the urinary odor concentration decreased, with this decrease being significantly greater while GDX females received EB. Thus, estradiol enhanced detection thresholds for male urine in a sex-specific manner, and this enhanced sensitivity in females was correlated with altered odor-sampling behavior.     

Mediation of male mouse urine marking and aggression by the vomeronasal organ

The effects of vomeronasal organ removal (VNX) on male mouse urine marking and aggressive behaviors were investigated. In three different stimulus conditions VNX male marking rates were about half that of sham-operated males. Aggressive behavior was tested by pairing males with male-urine-swabbed castrate males. Only 1 of the 12 VNX males displayed normal levels of fighting behavior and 6 did not initiate any fights during the aggression tests. These results indicate that normal male aggressive and urine marking behaviors are dependent on the presence of an intact vomeronasal system for their expression.     

Sexual motivation is demasculinized, but not feminized, in prenatally stressed male rats

Sexual motivation and copulation in male rats are associated with dopamine release in the nucleus accumbens. Demasculinized copulatory behavior has been demonstrated in prenatally stressed adult male rats. We have previously reported that approximately 80% of prenatally stressed male rats do not exhibit copulation and that no significant changes in nucleus accumbens dopamine release are seen during exposure to estrous females. In the present study, we investigated whether prenatal stress affects sexual motivation in these animals as adults. Pregnant Wistar rats were subjected to immobilization stress for two hours daily from day 15-19 of gestation. The prenatally stressed male offspring at the age of 3 months were allowed contact with receptive female rats for a 30 min period per week for 10 weeks; then, between the age of 5 and 6 months, their sexual motivation and copulatory activity were measured. Sexual motivation was measured in terms of sexual partner preference. The number of visits and the duration of each visit to an estrous female (stimulus female) or to a sexually active male rat (stimulus male) were recorded. Compared with control males, prenatally stressed male rats showed a significantly lower number of visits and a shorter duration of each visit to stimulus females. Prenatally stressed males showed no preference for male or female stimulus rats in terms of the number of visits and the duration of each visit, whereas control rats showed a significantly higher number of visits and duration of visits to female stimulus rats than male stimulus rats. A significant decrease in copulatory activity was observed in the prenatally stressed male offspring compared with control male rats, with most of the prenatally stressed males failing to show copulation. In vivo microdialysis experiments were performed on the nucleus accumbens with concurrent observation of sexual behavior. The prenatally stressed rats that did not exhibit copulation showed no significant changes in nucleus accumbens dopamine release during exposure to a stimulus male behind a wire-mesh barrier and the amount of dopamine release remained at the basal levels during actual physical contact. These results, combined with those of our previous report, indicate that sexual motivation in prenatally stressed male rats is demasculinized, but not feminized.     

Neuroendocrine control of urine-marking behavior in male rats

Sexually experienced Wistar male rats were used to investigate (a) urine voiding in the presence of nearby estrous females and the control of such voiding by (b) steroid hormones and (c) peripheral nerves supplying the genitourinary system. The first experiment showed that males always have a low rate of urine voiding that is significantly increased when a receptive female is around. Thus, it is suggested that an airborne scent from the female stimulates the olfactory system of males, triggering urine emission to transmit sex-related messages, i.e., male rats display the well-known urine-marking behavior of mammals. The number of urine marks and sniffing to females decreased after castration, and were restored after exogenous treatment with testosterone or estradiol. The proposed hypothesis is that airborne scents from the female activate the aromatization process in nuclei of the olfactory pathway of the male, evoking a cascade of neuronal responses that finish in urine marking. Peripheral nerves supplying the genitourinary system are the viscerocutaneous branch of the pelvic nerve (Vc) and the hypogastric (Hg). Data showed that both nerves are important for the central control of urine storage and voiding. Transection of Vc almost blocked urine marking, while Hg lesion increased the number of marks. Thus, it is discussed that Vc is the most important nerve in charge of voiding the bladder, and that Hg is important for continence.