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.     

Facilitory effects of male urine on feminine behavior in the male rat: Androgen-dependency

Feminine behavior in the male rat can be enhanced by exposing ORCH animals primed with estradiol benzoate and progesterone to the odor of urine collected from intact male adult congeners [20]. The present study provides evidence that this pheromonal effect is androgen dependent. A higher proportion of ORCH feminized rats displayed lordosis behavior following exposure to the odor of urine for either intact male rats or from ORCH rats supplemented with testosterone propionate than did ORCH feminized rats exposed to the odor of urine originating from non injected ORCH males.     

Olfactory environment and early mating behavior in the cyclic female rat

The objectives of this study were to examine the effects of olfactory cues from the male and of olfactory bulb removal on early mating behavior in sexually inexperienced diestrous female rats primed with estrogen. Four-day cyclic rats isolated from the male were given either 2.5 micrograms or 10 micrograms estradiol benzoate (EB) and presented to stimulated males in the late afternoon of diestrus 2 between 18:00 and 19:00 for a 10 min sexual behavioral session. Dose-dependent effects of estrogen were observed since 10 micrograms EB significantly increased the proportion of females displaying early mating as compared with those given 2.5 micrograms EB. Olfactory bulb removal prior to estrogen treatment caused a rise in the number of females which mated early with respect to the non bulbectomized controls. Exposing the females to bedding soiled with male urine on diestrus 2 at 10:00 did not affect early mating behavior. By contrast the olfactory stimuli became efficient when 2.5 micrograms EB treated females were given 10 micrograms progesterone (P) by the time of exposure to male urine. The results were discussed with respect to the role played by the olfactory system in the control of lordosis behavior throughout estrous cycle in female rats. P was concluded to be involved in the perception of the olfactory signals from the male which facilitate early mating behavior in diestrous female rats.     

Sexual behavior of male and female septal lesioned rats

Gonadectomized male and female rats were given septal lesions (SL) or sham surgery at approximately 60 days of age. After 3 weeks lordosis behavior tests were initiated. Females were tested after daily injections of 2 μg estradiol benzoate (EB) for 3 days, while males were tested after EB only (2 μg×3 days), and after EB plus progesterone (Prog). The mean lordosis quotients (LQ) of septal lesioned female rats were significantly higher than those of sham operated controls. No increase in lordosis responding was seen in male rats with either EB alone or EB+Prog. Following an additional 3 week interval without steroid treatment masculine behavior tests began. All animals received a pretest and were tested again on Day 4, 7, 11 and 15 daily tesosterone propionate (150 μg/day) treatment. No alterations in masculine sexual behavior (relative to that of controls) were found in either male or female septal lesioned rats. It is concluded that the increased hormone sensitivity is specific for lordosis behavior, at least when the SL are given in adulthood.     

Intrahypothalamic implants of noradrenergic antagonists disrupt lordosis behavior in female rats

There is considerable experimental evidence that hormonal activation of lordosis in female rats involves norepinephrine (NE) neurotransmission. However, no clear picture has emerged regarding either: 1) the neural sites at which NE influences lordosis, or 2) the NE receptor subtype(s) mediating NE effects on lordosis. To address these two issues, the behavioral effects of antagonists with relative specificity for alpha 1, alpha 2, or beta adrenergic receptors were examined. Drugs were administered via bilateral crystalline implants directly into the ventromedial nucleus of the hypothalamus (VMN) or medial preoptic area (MPOA) of ovariectomized female rats primed for 48 hr with 3 micrograms of estradiol benzoate (EB) and given 200 micrograms of progesterone (P) 3.5-4 hr before testing. When applied to the VMN 1 hr before the P injection, the alpha 1 receptor antagonist prazosin reduced lordosis behavior in 86% of animals but in only 10% of rats when applied to the MPOA. However, prazosin did not inhibit lordosis when implanted into the VMN just prior to EB administration. Yohimbine, an alpha 2 receptor antagonist with low affinity for alpha 1 receptors, also suppressed lordosis in 41% of animals with VMN implants and in 37% of rats with MPOA implants. By contrast, the alpha 2 antagonist idazoxan, which has little activity at alpha 1 receptors, did not significantly affect estrous responding when implanted into either the VMN or MPOA. VMN implants of the beta receptor antagonists propranolol and pindolol reduced lordosis behavior in 50% and 86% of rats, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in spontaneous,odor modulated and shock induced behavior patterns following discrete olfactory system lesions

Three groups of male hooded rats were preoperatively matched and then sham operated (SHAM), bilaterally lateral olfactory tract lesioned or anterior olfactory nucleus/anterior commissure lesioned. In one experiment, spontaneous behavior patterns emitted in an exploration field containing different odors were quantified during satiated and food deprived conditions. In other experiments, flinch and jump thresholds to electric shock and running patterns in an appetitively motivated straight alley experiment were measured. All lesioned animals were hyperactive and typically froze and groomed less than SHAMs while sniffing and rearing patterns differentiated the lesioned and SHAM groups in several ways. All animals had similar flinch and jump thresholds and also emitted similar types of responses to electric shock. In the straight alley, odors from normal and stressed rats had little effect on the running time of the experimental animals while a faint cat odor strongly inhibited SHAM running behavior. Responses to a novel chemical odor (trimethylpentane) and to changes in the alley floor or electric shock applied to the floor were minimal in all animals. The results were discussed against a background of other olfactory system lesions and some evidence separating the effects of olfactory cues from nonolfactory lesion effects was presented. Finally, problems associated with tests for olfactory discrimination were discussed. A need for more rigorous and specific discrimination tests in behavioral studies involving anosmia was emphasized.     

Estradiol benzoate facilitates lordosis and ear wiggling of 4- to 6-day-old rats

The effects of exogenous and endogenous steroids on components of female sexual behavior of neonatal male and female rats were investigated. In Experiment 1, 4-day-old rats were treated with 0, 0.1, 1.0, 10, or 100 micrograms/10 g body weight estradiol benzoate (EB) and were tested 44 hr later. In Experiment 2, male rats castrated within 24 to 48 hr of birth were compared with sham operated controls and castrates given steroid replacement. The results indicated that most 6-day-old pups will display lordosis and ear wiggling; therefore, the display of these responses is not dependent upon exogenous steroids. However, a fine-grained behavioral analysis revealed that EB treatment increased the frequency, duration, and intensity of lordosis and the frequency of ear wiggling in infant females, and it increased lordosis duration in males. Castration of infant males decreased the likelihood that male infants would display lordosis, whereas testosterone replacement restored behavior to control levels. These data question the concept that organizational and activational actions of estrogens occur during completely separable times in development and should provide new insights into the development of estrogen receptor function and the process of sexual differentiation of brain and behavior.     

Lordosis inhibiting effects of endogenous progesterone in the male rat primed with estrogen

The aim of this study was to investigate the mechanisms involved in the inhibitory action of progesterone on estrogen-induced facilitatory effects of estradiol benzoate on lordosis behavior in the male rat. Intact adult male rats were given 1) 25 micrograms estradiol benzoate (EB) and 100 micrograms progesterone (P) at an interval of 42 hr. EB injected animals served as controls 2) EB followed by 3 doses of 400 micrograms dexamethasone (DEXA) and P as above. EB + DEXA injected animals served as controls. Testing for lordosis behavior was performed by 50 +/- hr after EB injection. A significant decrease in the number of the males displaying lordosis in response to the mounts of stimulus males resulted from P injection following EB treatment as compared to EB controls. DEXA treatment significantly reduced the number of EB animals showing lordosis responses but completely prevented the inhibitory effects of exogenous P to occur. Blood P values appeared to be significantly lower in EB + DEXA males than in their EB counterparts. The results provide evidence that endogenous P is involved in the display of lordosis behavior by EB-treated intact males. They mainly suggest that the effects of exogenous P on estrogen-induced lordosis behavior in the intact male rat result from sequential inhibitory mechanisms involving exposure of the animals to the successive action of endogenous and exogenous P.     

The role of mesencephalic tegmentum in regulating female rat sexual behaviors

Feminine sexual behaviors were tested in estradiol benzoate (EB) and progesterone (P) primed ovariectomized rats following four types of radiofrequency lesions in the midbrain tegmentum. The dorsomedial lesion (DML) which destroyed the ventromedial central gray including the dorsal raphe nucleus and adjacent area induced high sexual receptivity in the females primed with low dose (0.5 micrograms) of EB-P. All females with DML exhibited lordosis and ear wiggling, the mean lordosis quotient (LQ) being significantly higher than that of castrated controls or sham operated rats. Sexual receptivity in females with ventromedial tegmental lesion was not significantly different from those of the control and sham groups. In contrast to the medially lesioned groups, the mean LQ was low in the animals with bilateral lateral tegmental lesions even when the dose of EB was increased to 2 micrograms which was sufficient to induce high sexual receptivity in castrated and sham operated control females. In the animals with dorsolateral tegmental lesions (DLL), a much more severe loss of lordosis was seen than in those with ventrolateral tegmental lesions (VLL). None of the DLL females displayed sexual behavior throughout the present experiments. These results lead us to conclude that the midbrain dorsomedial tegmental area (ventral central gray and the adjacent area) is concerned with female sexual behavior inhibiting system, whereas the lateral tegmental area may be involved in the facilitatory system.     

Disruption of diurnal feeding and weight gain cycles in weanling rats by ventromedial and dorsomedial hypothalamic lesions

At the age of 28 days, male Sprague-Dawley rats received bilateral electrolytic lesions in the ventromedial (VMN) and dorsomedial (DMN) hypothalamic nuclei, repectively. Sham-operated rats served as controls. Food intake and body weight gains were measured for 13 days at the beginning of the light period (0800–2000 hr) and prior to the start of the dark period (2000-0800 hr). Both types of hypothalamic lesions caused a disruption of the naturally occurring diurnal feeding and weight gaining cycles. In accordance with previous data, the VMN rats remained normophagic and made normal weight gains while DMN animals were hypophagic and gained weight subnormally; linear growth was reduced in both types of lesioned animals, but only the VMN rats became obese. The data suggest the existence in the medial hypothalamus of an area that is involved in feeding and weight gaining cycles. From the standpoint of overall-caloric intake, this area consists of two types of neuronal assemblies that differ fuctionally as profoundly as they are anatomically separate.     

Olfactory,septal and amygdala lesions alone or in combination: Effects on lordosis behavior and emotionality

Ovariectomized female rats subjected to olfactory bulbectomy (OB) and treated with 0.5, 1.0 or 2.0 μg estradiol benzoate (EB) per day for three days showed a dose-related increase in lordosis behavior similar to that of septal lesioned (SL) rats, and relative to the response of sham operated controls. Animals with dual lesions (SL + OB) also showed increased behavioral sensitivity to estrogen, but the lesion effects were not additive. Rats given amygdala lesions (AL) showed levels of lordosis behavior comparable to that of sham animals following 2 μg EB per day for 3 days. However, AL attenuated the SL induced increase in lordosis behavior in animals given dual lesions (Sl + AL). All animals were tested for emotionality prior to surgery and on Postsurgical Days 1, 3, 7, 14, 21 and 42. Only the SL rats showed increased emotionality, and this increase was unrelated to the facilitation of lordosis behavior. It is suggested that OB and SL may exert their effects on lordosis behavior via a common neural pathway or system. Since AL reduced behavioral sensitivity to estrogen in SL rats, the amygdala may be a part of the neural system mediating the increased sensitivity to estrogen in SL rats.     

REM sleep deprivation facilitates the estrogen effect on heterotypical sexual behavior in male rats

Gonadectomized male rats were submitted to REM sleep deprivation (REMd) for 120 hr and their hetero and homotypical sexual response to estradiol benzoate (EB) was tested. Subjects (Ss) receiving 20 micrograms EB showed lordosis quotients (LQ) twice as high as those receiving 10 micrograms EB and at the same time the LQs in these groups were higher than in the non-REMd groups. Gonadectomized-adrenalectomized control Ss showed the highest levels of lordosis throughout the experiment. REMd by itself does not produce lordosis response. The results indicate that the brain structures underlying this behavior in males are probably similar to those in females, since REMd increases lordosis in both cases. The lack of homotypical sexual behavior normally observed in gonadectomized male rats does not seem to be affected by this treatment. The issue of adequate controls for REMd experiments is discussed.     

Olfactory cues and accessory olfactory bulb lesion: effects on sexual behavior in the cyclic female rat

The influence of olfactory cues and accessory olfactory bulb (AOB) lesion on female sexual behavior was studied in virgin female Wistar rats. In Experiment 1, it appeared that distance or contact exposure to male urine soiled bedding for 8 hours before testing increased sexual receptivity, i.e., the number of receptive females at 18:00-19:00 on proestrus. In Experiment 2, we observed that sexual receptivity at 18:00-19:00 on proestrus was not affected by AOB lesion as compared to sham-operated females. In Experiment 3 the effects of both AOB lesion and olfactory cues were analyzed. Sexual receptivity at 18:00-19:00 on proestrus did not significantly differ in sham-operated and accessory olfactory bulbectomized females both exposed to the odor of male urine. Regarding lordosis quotient in the three experiments, no significant difference was observed. Mechanisms whereby olfactory cues and/or AOB lesion modified female sexual behavior on proestrus in virgin female rats were discussed in the light of previous and present observations.     

Hypothalamic and midbrain control of sexual receptivity in the female rat

The ventromedial nucleus of the hypothalamus (VMN) appears to be one site of action for estrogen with respect to the induction of sexual receptivity. Lateral efferents from the VMN pass through a region capping the lateral part of the cerebral peduncle and constitute a “lateral pathway” connecting the VMN with midbrain structures. Bilateral lesions of this peripeduncular region markedly reduced the display of lordosis by estrogen or estrogen-progesterone primed female rats in sex behavior tests with males. Sexual receptivity was comparably reduced when a sagittal knife-cut lateral to the VMN on one side of the brain was combined with a lesion of the peripeduncular region on the other side of the brain. In other words, asymmetric brain damage which nevertheless bilaterally interrupts axons of the lateral pathway connecting the VMN and midbrain markedly reduces the occurrence of lordosis, and we conclude that the integrity of the lateral pathway is essential for the hormonal induction of sexual receptivity in female rats. This pathway may be one avenue through which the action of estrogen on cells of the VMN may bias activity in midbrain structures directly concerned with the elicitation and/or execution of the lordosis reflex, to bring about the occurrence of sexual receptivity in the female.     

Naloxone suppression of food and water intake and cholecystokinin reduction of feeding is attenuated in weanling rats with dorsomedial hypothalamic lesions

In Experiment 1, sham operated (SCON) and dorsomedial hypothalamic nuclei (DMN) lesioned (L) rats were given saline or naloxone (0.1, 1.0 or 2.0 mg/kg) just prior to the onset of the dark cycle, lights out. Compared to saline injections, naloxone at all doses suppressed the cumulative food intake of the SCON during the second and third hr of measurement. Naloxone was without significant effect on the food intake of DMNL rats. Similar results were obtained in Experiment 2, except that naloxone at 2.0 mg/kg significantly suppressed the DMNL rats' food intake by the fourth hr of measurement. Cumulative water intake of both groups was significantly suppressed by naloxone in both experiments but its effects appeared to be attenuated in the DMNL group. In a preliminary trial cholecystokinin octapeptide (3.0 and 6.0 micrograms/kg) given at the onset of the dark cycle significantly suppressed the food intake of the SCON group but had no significant effect on the DMNL rats. The possibility exists that the reduced food intake and lower body weight of DMNL rats may partially result from damage to an opioid system. The data also tentatively suggest that DMN may play a role in cholecystokinin-induced satiety.     

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.     

Effects of stimulus female on sexual behavior of male rats given olfactory tubercle and corticomedial amygdaloid lesions

Lesions of the olfactory tubercle (OT) or corticomedial amygdala (CMA) damaging projections from the main or accessory olfactory bulbs, respectively, were made in male rats. When paired with ovariectomized females treated with estradiol benzoate (EB) and progesterone (Experiment 1) sexual behavior of lesioned males did not differ from sham and unoperated controls. During Experiment 2 males were paired with females chronically treated with EB only. Lesions of the OT did not affect mating while CMA lesions produced lengthened ejaculatory latencies (EL), longer mean interintromission intervals and more intromissions per ejaculation. Lesions of the CMA produced EL which were either within a normal range or noticeably extended with ejaculatory deficits becoming more exaggerated through three ejaculatory series. Low levels of soliciting by EB treated females seemed to precipitate ejaculatory deficits.     

Dissociation between the display of lordosis and soliciting behaviors in female rats with lesions of the dorsomedial pontine tegmentum

Lesions of the dorsomedial tegmentum (DMTL) between the midbrain pontine junction and the middle level of the pons effectively eliminate the induction by estrogen-progesterone of lordosis behavior in ovariectomized rats. However, soliciting behaviors such as ear wiggling and hopping were not inhibited by this type of lesion. The common damaged area in DMTL rats which failed to show lordosis was the medial periventricular gray. The lesions placed in the caudal pontine central gray were not effective in suppressing lordosis response. Lesions of the ventromedial tegmentum (VMTL) were also ineffective in suppressing lordosis. Most of the animals with the VMTL showed soliciting behaviors. In these rats, the incidence of lordosis and lordosis quotient (LQ) were comparable to those of sham operated rats. When bilateral lesions were placed in the lateral tegmentum region, the mean LQ and incidence of soliciting behavior were not significantly different from those of sham operated controls. These results suggest a clear dissociation of the regulatory mechanisms between the display of lordosis and soliciting behaviors at the pontine level.     

Ultrastructural evidence for enkephalin mediated disinhibition in the ventromedial nucleus of the hypothalamus

The ventromedial nucleus of the hypothalamus (VMN) regulates the estrogen-dependent appearance of female mating behavior, lordosis. Accumulating evidence suggests that estrogen might exert its control over lordosis by acting, in part, on neurons that contain enkephalin in the VMN. The expression of the enkephalin precursor gene is robustly stimulated by estrogen and is correlated with the later appearance of lordosis. GABA has also been implicated as an important neurotransmitter for the appearance of lordosis. Because enkephalin is thought to act in several brain areas to modulate the activity of GABAergic neurons, we studied the ultrastructural morphology and relationship between neurons containing these neurochemicals using dual-labeling immunocytochemistry in ovariectornized rats, half of which received estrogen replacement. Immunolabeling for enkephalin was almost always detected within axon terminals (695 axonal profiles sampled), while GABA immunoreactivity was more often localized to cell bodies and dendrites (191 profiles), than to axons (63 profiles). Axon terminals containing enkephalin immunolabeling provided a major innervation to soma or dendrites containing GABA. That is, over one third (94/245) of the axon terminals in contact with GABA-immunoreactive dendrites contained enkephalin. Furthermore, these GABA-immunoreactive dendrites accounted for a fifth of the somatodendritic processes associated with enkephalin-containing axon terminals. These findings support the hypothesis that enkephalin may act in the VMN by inhibiting GABAergic neurons, which could result in the disinhibition of neural circuits relevant for lordosis.     

Estrous odors and sexually conditioned neutral odors activate separate neural pathways in the male rat

Olfactory stimuli play important roles in sexual behavior. Previous studies have demonstrated that both estrous odors and initially neutral odors paired with copulation influence the sexual behavior of male rats. The present study examines the pattern of neural activation as revealed by Fos immunoreactivity (Fos-IR) following exposure to bedding scented with either a neutral odor (almond) paired previously with copulation, estrous odors or no odor. Following exposure to estrous odors Fos-IR increased in the accessory olfactory bulb, medial amygdala, medial bed nucleus of the stria terminalis, medial preoptic area, ventromedial hypothalamus, ventral tegmental area, and both the nucleus accumbens core and shell. Conversely, following exposure to the sexually conditioned odor Fos-IR increased in the piriform cortex, basolateral amygdala, nucleus accumbens core, and the anterior portion of the lateral hypothalamic area. In addition, following exposure to almond odor Fos-IR increased in the main olfactory bulb independent of its pairing with copulation. These patterns of Fos-IR following exposure to estrous or sexually conditioned odors were not influenced by either the addition or omission of the other type of odor. These findings demonstrate that estrous and sexually conditioned odors are processed by distinct neural pathways and converge in the nucleus accumbens core, suggesting that this structure has a unique role in processing sexual stimuli of both pheromonal and olfactory natures.