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Ultrasonic vocalizations were tape recorded from five pairs of copulating mice and subjected to spectrographic analysis. As expected, the mice emitted numerous 70-kHz vocalizations. At the beginning of the test, before copulation began, 70-kHz calls were emitted almost continuously, while calls with lower spectrographic frequencies were not observed. Subsequently, bursts of 70-kHz calling generally began shortly before mounts and intromissions and persisted until dismount. Intermixed with these 70-kHz calls were additional vocalizations of about 40 kHz. Calling rates were highest just prior to intromission. Once intromissions began, 70-kHz calls continued at a lower rate until dismount; however, 40-kHz calls occurred infrequently. In a second experiment, the male was found to emit the majority of the 70-kHz calls and all of the 40-kHz calls. When the male was devocalized, few calls were detected, regardless of whether the female was able to call. If the male was not devocalized, high rates of calling were detected, even if the female was devocalized.

Copulatory behavior and associated social behaviors such as ultrasonic vocalizations and scent marking are reduced in frequency following castration and are restored by exogenous administration of androgens. In the present study, we report the behavior of a subgroup of male Long-Evans rats in which there was a dissociation between the responses of androgen-dependent behaviors to castration. Five weeks after castration, 52% of the males tested (13 of 25) had higher 50 kHz vocalization frequencies than during precastration tests. This group continued to emit vocalizations after castration and actually increased their number of vocalizations over postcastration tests (pretest: 34.5 +/- 4.8 to week 15: 62.8 +/- 10.9/10 min test). The remaining males (n = 12) exhibited a decline in vocalizations (pretest: 29.7 +/- 5.1 to week 15: 6.5 +/- 2.7) that we typically observe in our laboratory. Both groups showed the expected decline in scent-marking frequency over the postcastration tests and were impaired in performance of copulatory behavior. Seminal vesicle and adrenal gland weights did not differ between the two groups. The 25 males were of the same genetic strain as previous animals in our laboratory except that they were born and raised in a different location prior to shipment to our laboratory. Androgens, therefore, may be only one of the possible influences mediating ultrasonic vocalizations.

Castrated male mice were bilaterally implanted with 27 ga cannulae containing testosterone into either the septum, medial preoptic area (MPO), or corticomedial amygdala. One additional group of castrates received no hormone and another received only systemic testosterone via subcutaneous silastic capsules. All males were subsequently tested for ultrasonic mating vocalizations, urine marking, mounting behavior, aggression and gender preference, all of which are androgen-dependent, male-typical behaviors. In general castrates receiving no hormone performed these behaviors at low levels and animals receiving systemic testosterone performed the behaviors at normal male-typical levels. Ultrasonic vocalizations in response to female urine were activated by MPO implants. Urine marking in response to female urine appeared to be partially activated only with MPO implants. Very little mounting or fighting were seen in the brain implanted groups. Gender preference (for females over males) was restored with MPO implants and appeared to be partially activated with septal implants. The seminal vesicles of the castrates receiving brain implants were not significantly different from those receiving no hormone indicating that little or no implanted hormone was exiting the brain into general circulation. The implications of these findings for the neuroanatomy of sexual motivation and performance are discussed.

RU 486 is known primarily as an antagonist to progestins and glucocorticoids. However, RU 486 has also been shown to have agonistic progestational properties in biochemical and behavioral studies. In the current study, RU 486 was implanted directly into the ventromedial hypothalamus (VMH) to test for facilitative action on the receptive behavior of female ovariectomized Long-Evans rats primed with 5 micrograms of estradiol benzoate. Cannulae containing RU 486, progesterone (P), or empty cannulae were implanted 48 hr after estrogen priming. The lordosis quotient and the lordosis score were assessed 4 hr after the cannulae were lowered by a standardized test consisting of 10 mounts by a stimulus male. P and RU 486 significantly facilitated receptivity compared to blank implants in terms of lordosis quotient and lordosis score, with no significant difference between the hormone treatments. While only a single dose of each treatment was given in the current study, RU 486 facilitated lordosis when implanted to the VMH as well as progesterone in contrast to our previous results where the steroids were administered systemically.

Female rats emit calls during copulation that appear to be associated with their own proceptive behavior. In an earlier study, we had determined that females emit calls when presented with odor cues from a sexually competent adult male. Two experiments were undertaken to determine whether females would vocalize when presented with tape recorded male ultrasonic vocalizations. In the first females were presented with no calls, male mating calls or male preejaculatory calls. They received one test in each sound condition with a devocalized male in the immediate vicinity and one test with no male present. Females tested alone called at a lower level than those tested in the presence of a devocalized male. The presence of male calls did not significantly increase the frequency of calling in females tested in the presence of a male. When females were tested alone, however, calling did increase significantly when male mating calls were presented, but not when preejaculatory calls were presented. In a second experiment, it was confirmed that male mating vocalizations elicited more calls from the female. Thus, male mating calls may elicit vocalizations from the female, but are less effective than male odor cues.

Two experiments examined the role of the steroid hormones, estradiol (E2), progesterone (P), and testosterone (T), in activating scent marking and 50-kHz ultrasonic vocalizations in ovariectomized Long-Evans rats in response to a devocalized male rat. In Experiment 1, females received, in a counterbalanced order, a series of six hormone treatments consisting of two injections (48-54 and 4 hr before behavioral tests). The six treatments were 8 micrograms E2 followed by 500 micrograms P or oil, 2 micrograms E2 followed by 500 micrograms P or oil, and oil followed by 500 micrograms P or oil. Injections of either the high or low dose of E2 followed by P resulted in high levels of vocalizations. Neither E2 by itself or P by itself were very effective. Surprisingly, none of the hormone treatments were effective in activating marking above the level seen when the females received control injections of oil. Four other hormone treatments were examined in Experiment 2: daily injections of 500 micrograms T, daily injections of 50 micrograms E2, implantation of silastic capsules of E2 (5% E2, 5 mm length) followed by 500-micrograms P injections before behavioral tests, and implantation of silastic capsules of E2 followed by oil injections. Animals receiving E2 by silastic capsule followed by P injection displayed the highest levels of marking and vocalizations across the five weekly tests. These results suggest that while E2 and P synergize for the display of female-typical behaviors similar to the hormonal regulation of lordosis, the mechanism of E2 action may be different for the two signaling behaviors. Scent marking appears to be responsive to the tonic levels of E2 released from silastic capsules.

The purpose of the present study was to investigate whether the signaling behaviors of female Long-Evans rats varies over the estrous cycle. Scent marking and 50-kHz ultrasonic mating vocalizations, in response to a devocalized sexually experienced male, changed in frequency across the cycle, both behaviors being highest at proestrus/early estrus. Vocalizations were recorded on every day of the cycle and also in females showing no regular cycles. In contrast, scent marking was rarely observed during the estrous cycle except at proestrus/early estrus. These results suggest that both behaviors, which increase around the time of sexual receptivity, may help to coordinate reproduction and further suggest an endocrine basis for the changes observed over the estrous cycle.

Two experiments in house mice (Mus domesticus) examined the neural sites at which steroid hormones activate the following male-typical behaviors: 70 kHz ultrasonic mating vocalizations in response to stimulus females or their urine, urinary marking in response to stimulus males or stimulus females, mounting of estrous females, and intermale aggression. In the first experiment, four groups of castrated males received bilateral intracranial implants of testosterone (T) into either the septum (SEPTUM), medial preoptic area (MPO), anterior hypothalamus (AHA), or ventromedial hypothalamus (VMH). Two control groups received subcutaneous silastic capsules of T (TSIL) or empty silastic capsules (BSIL). The TSIL males performed all behaviors at male-typical levels while the BSIL males were unresponsive. MPO males emitted ultrasonic mating vocalizations at high levels while few vocalizations were seen in males of the other brain implant groups. The VMH, AHA, and MPO males urine marked at higher levels than the BSIL males but did not exhibit the high levels of the TSIL males. Mounting was observed only in MPO and TSIL males. Aggression was rare in males from any of the brain implant groups. In the second experiment, the hormone activity of the implants was increased by using testosterone propionate (TP) or a 50% mixture of estradiol (E2) and cholesterol. The six groups were SEPTUMTP, SEPTUME2, MPOTP, MPOE2, TPSIL, and BSIL. The TPSIL males performed all behaviors at male-typical levels while the BSIL males were unresponsive. TP was effective at restoring vocalizations and urine marking only when placed in the MPO; however, E2 was effective at both sites. Again aggression and mounting were less evident in the brain implanted males. In conclusion, implants of T or TP were effective in restoring ultrasonic mating vocalization when placed into the MPO. MPO implants of T and TP were also effective in stimulating urine marking, although VMH and AHA implants also showed some effectiveness. The restorative effects of E2 were not localized which is probably related to the greater hormonal activity of this treatment. Comparisons of the properties of the various brain implants to restore more than one behavior were discussed.

The effects of testosterone (T) and its major metabolites, estradiol (E2) and dihydrotestosterone (DHT), on the restoration of sebaceous scent marking and 50-kHz ultrasonic vocalizations in male rats were measured in two studies (Experiments 1 and 2) employing different hormone levels. Silastic capsule administration of high and low doses of T (10 and 20 mm) or E2 (5%; 5 and 10 mm) completely restored marking to precastration levels. Both doses of DHT (30 and 40 mm) or no hormone replacement were without effect when tested in presence of estrous odor cues. In our testing paradigm, males appeared to mark glass objects with a sebaceous secretion rather than urine. Only the high dose of T (20 mm) restored vocalizations to intact levels, while animals receiving the high or low dose of E2 or DHT showed no restoration of behavior. In Experiment 3, habituation to estrous odor cues over weekly tests was not observed for marking or vocalizations. These results are discussed with respect to similarities and differences in the hormonal control of signaling behaviors by androgen metabolites in male rodents.

Progesterone (P) facilitation of sexual receptivity in rodents has been achieved by intracranial administration to the ventral hypothalamus; the preoptic area; and midbrain areas such as central gray, mesencephalic reticular formation, and ventral tegmental nucleus. In our laboratory, by far the most effective site in rats has been the ventromedial nucleus of the hypothalamus (VMN). However, several reports of sensitivity to P in the midbrain of rats and other rodent species led us to investigate whether stimulation of the ventral midbrain of female rats might contribute to facilitation of sexual receptivity. Ovariectomized Long-Evans rats received one cannula aimed at the VMN, and another aimed at the contralateral ventral mesencephalon. P in both cannulae, following a priming dose of estradiol, caused significantly higher lordosis quotients (LQ) than blank tubes. Controls with bilateral cannulae in the VMN responded when both tubes were filled with P, but did not respond to unilateral VMN P stimulation. P in the VMN and contralateral anterior preoptic area did not result in a greater degree of receptivity than did the empty tubes. These studies indicate that although progesterone stimulation in the midbrain alone is not sufficient to facilitate receptivity in female rats with our methods, the midbrain may play an auxiliary role. P implants in the midbrain appear to facilitate receptivity in the case of VMN implant treatments that are subthreshold for stimulating lordosis. The results are discussed in light of similar studies in other rodent species, and in the context that more than one brain site may be important in the natural stimulation of sexual receptivity by gonadal hormones.

Female rats emit ultrasonic vocalizations during copulation. Female vocalizations are associated with darting and other proceptive behavior. In addition, females frequently call while approaching the male. A series of experiments was undertaken to determine whether female rats emit ultrasonic vocalizations in response to other rats or to their odors. Females were exposed either to bedding soiled by males or females or to devocalized conspecifics separated from the subject by a barrier. Females vocalized more to male cues than to female cues, regardless of whether the odor cues were from soiled bedding or from a devocalized conspecific. In addition, subjects vocalized more when presented with a devocalized female rather than a neutral stimulus. Furthermore, devocalized adult males, separated by a barrier, were more effective than either devocalized castrated or juvenile males in eliciting vocalizations; in turn, castrates and juveniles were more effective than neutral stimuli. Female calling was likely induced by odors from the male. In one experiment, the female subject was positioned in such a way that she could neither see or touch the stimulus male. Presentation of tape-recorded male vocalizations did not affect calling by the female. Vocalizations emitted by females in response to male odors may attract males or may facilitate subsequent copulatory behavior by the male.

The progestin receptor antagonist RU 38486 (henceforth referred to as RU 486) was tested for facilitative effects on female receptive behavior in ovariectomized Long-Evans rats primed with 2 micrograms estradiol benzoate (EB). RU 486 (0, 0.5, 1.6, or 5.0 mg) was administered 48 hr after estrogen priming. The lordosis quotient (LQ) and lordosis score (LS) were assessed 4 hr after RU 486 administration in a standardized test consisting of a 10-mount test by a stimulus male. A significant dose effect was found by both LQ and LS, with those subjects receiving 5 mg of RU 486 being significantly more receptive than vehicle control animals. Thus RU 486 acted as a weak progestin agonist under testing conditions typical for assessment of progestin facilitation of female sexual behavior in rats. Low levels of proceptive behavior (hops and darts) were seen in a minority of the tests, and did not vary systematically as a function of the dose of RU 486 administered. We also examined the effects of RU 486 given before progesterone (P) on receptivity in a blocking paradigm and confirmed previous reports that the antagonist significantly attenuates facilitation of sexual behavior when given in combination with P. A progestin receptor assay of the cytosols of the hypothalamus-preoptic area in estrogen-primed female rats treated with 5 mg RU 486 revealed a significantly greater depletion of available cytosolic P receptors than when rats were treated with a similarly facilitating dose of P (100 micrograms). The results suggest a possible dual mode of action for RU 486--a weak, receptor-mediated agonistic effect on sexual behavior when given alone to estrogen-primed rats, and a competitive blocking effect on receptivity when administered with P.

Extracellular dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) in the nucleus accumbens (N. Acc.) were measured by in vivo microdialysis during male sexual activity. DA and DOPAC were significantly increased during copulation, but not during mild tail pinch. These results are consistent with studies showing increases in N. Acc. DA associated with positively reinforcing environmental stimuli.

Both male and female rats (Rattus norvegicus) emit ultrasounds during copulation. Bursts of ultrasounds from males that occur as ejaculation approaches are lower in frequency, longer in duration, and louder than those that appear earlier in the ejaculatory series. We attempted to determine if these pre-ejaculatory calls affect female sexual behavior. The behavior of females paired with devocalized males was compared with that of females paired with intact males, and in a second experiment tape-recorded vocalizations were played to females paired with devocalized males. Females were less receptive when paired with devocalized males; they were less likely to remain stationary when males attempted to mount. Playback of both types of calls restored female immobility toward control levels, although pre-ejaculatory calls were more effective than mating ones. Pre-ejaculatory calls restored running and training levels somewhat toward control level but to a lesser extent than female immobility.

Male rats (Rattus norvegicus) emit at least two patterns of vocalization during copulation, the mating call and the pre-ejaculatory call. Both calls promote immobility of the female during lordosis, but the pre-ejaculatory calls are more effective. We undertook, through ventral observations of the mating pair, to determine if the female failed to assume or maintain the lordosis posture when mounted by a devocalized male and also to determine if the devocalized male was providing adequate stimulation to induce receptive behavior. Females were more likely to move away from the devocalized males before assuming the full lordosis posture. Furthermore, they were more likely to move away before the males had a chance to engage in intromissive behavior. However, when the females remained immobile long enough for the males to achieve a mount or intromission, there was little difference in the behavior of either animal that resulted from the devocalization of the male.

The present study was carried out in order to assess the time course of action of progesterone (P) in the facilitation of complete feminine sexual behavior. Female rats (estrogen primed via 5% E2 Silastic capsules) were given 200 micrograms of P either intravenously (iv) or subcutaneously (sc), and tested for estrous behavior at 1/4, 1/2, 1, 2, and 4 hr after treatment. Among iv-treated animals, significant amounts of lordosis behavior were seen as early as 1/2 hr, and a dramatic rise in solicitation behavior was observed at 2 hr. Although sc-treated animals displayed significant amounts of lordosis and solicitation behavior at 2 hr, the behavior was not maximal until 4 hr. Intravenous administration of 400 micrograms P was equipotent to 200 micrograms P, whereas 50 micrograms of iv P was relatively ineffective. A dual mechanism hypothesis pertaining to progesterone's actions in the facilitation of both the receptive and proceptive components of feminine sexual behavior in rats is discussed.

The CNS sites of action for progesterone facilitation of female sexual behavior are disputed. Among the areas most often cited are the ventromedial hypothalamus and the ventral midbrain. There is also a controversy about whether estradiol may substitute for progesterone for the facilitation of receptive behavior when given systemically or intracranially. We tested VMH and ventral midbrain applications of estradiol versus progesterone for the facilitation of female sexual behavior in estrogen-primed, ovariectomized female rats. Subjects were implanted with bilateral guide tubes aimed at ventral hypothalamic or midbrain sites. Estrogen-primed rats received either 28-gauge insert cannulae filled at the lumen with pure progesterone, estradiol, or cholesterol, or empty tubes, and were tested for receptivity with intact, experienced stud males just before, and 1 and 4 hr after, intracranial hormone administration. Significant estrous responsiveness was seen only in the 4-hr test after progesterone was implanted in the VMN in the first intracranial cannula test. We conclude, in contrast to some previous reports, that administration of progesterone to the VMN is more effective in the facilitation of female sexual behavior than when it is implanted in the ventral midbrain, and that administration of estradiol to either site is ineffective.

The present experiment was designed to determine whether sequential replacement of estrogen and progesterone to the ventromedial hypothalamus (VMH) would be sufficient to induce estrous behavior in ovariectomized rats. Bilateral cannulae containing 17 beta-estradiol (E2) diluted with cholesterol (1:250) were lowered into the VMH, preoptic area or midbrain and left in place for 4 days. On day 5, the E2 inserts were removed and P-filled cannulae were lowered into half of the subjects. The remaining females received systemic progesterone (500 micrograms). This steroid regimen was repeated 2 weeks later with the mode of progesterone administration reversed. All subjects were tested for estrous behavior twice after progesterone treatment. In a second experiment, 3H-P:P-filled cannulae were lowered into the VMH of estrogen-primed females in order to estimate the extent of hormone spread from full-strength P-filled cannulae. Results indicated that estrogen and progesterone stimulation of the VMH is sufficient to activate estrous behavior in spayed female rats, however, precise localization of the hormone implants within the VMH is essential. 9 of the 11 females with both cannulae located within or at the border of the ventromedial nucleus (VMN) exhibited estrous behavior whereas only half of the females with only one implant resting in the VMN exhibited estrous responsiveness. Subjects with neither cannula located within or at the border of the VMN did not exhibit the behavior. The facilitative effects of P appeared to result from hormonal stimulation of the VMH and not from leakage of the steroid into other brain regions or into the systemic circulation. Following placement of tritiated progesterone implants into the VMH, high levels of radioactivity were recovered only from the mediobasal hypothalamus. The low levels of radioactivity measured in other brain regions, pituitary, uterus and blood indicate that relatively little if any hormone reached these tissues.

The hypothesis that progesterone (P) activates estrous behavior in estrogen-primed female rats via a protein synthetic mechanism was examined. The protein synthesis inhibitor anisomycin was applied intracerebrally via 28-gauge bilateral implants to neural sites implicated in the mediation of estrous responsiveness. Results showed that anisomycin blockade of P-activated estrous behavior was neural site specific. Animals with anisomycin placed in the ventromedial hypothalamus (VMH) (n = 27) showed low levels of lordosis and solicitation behavior 4 h after the subcutaneous administration of 500 micrograms P, whereas animals with anisomycin implants in the preoptic area (n = 11) or the midbrain area in the region of the interpeduncular nucleus (n = 11) displayed high levels of estrous behavior. No deficits in open-field activity were observed following localized anisomycin treatment, and all animals appeared to be healthy. Results of the present study are consistent with the hypothesis that P acts to promote estrous behavior via a receptor-mediated genomic protein synthetic mechanism, and provide additional evidence that the VMH is the primary site of P action.

Ultrasonic vocalizations were measured when male rats were placed with ovariectomized females that had experienced various hormonal and behavioral treatments. In Experiment 1, 18 males were tested with females in each of the following conditions: nonestrous (OVX), estrogen treated (E), estrogen and progesterone treated (EP), and estrogen and progesterone treated and given two intromissions from a stud male prior to testing (EPI). Control conditions included clean cage (CL) and cage soiled by an estrous female (SOI). The treatments differed in effect on rate and maintenance of vocalization: EP greater than E greater than EPI = OVX greater than SOI greater than CL. In tests in which males produced a high rate of vocalization, some males with short intromission latencies shifted from the normal 50-kHz pulse to a 22-kHz pulse. In Experiment 2, the effect of the female's vocalization and movement on the rate of and latency to vocalization was measured. Twenty-one males were presented with each of the following stimulus conditions: estrous female with red light (EP), estrous female without red light (EP dark), estrous anesthetized female (EP anes), and nonestrous anesthetized female (OVX anes). Effects on vocalization of various treatments were as follows: EP = EP dark greater than EP anes greater than OVX anes. These data suggest that the 50-kHz vocalizations constitute a graded response influenced by the female's hormonal and sexual condition.

Females exposed to morphine sulfate in utero (5-10 mg/kg twice a day on days 11-18 of gestation) displayed precocious vaginal opening and had increased body weight from the 8th week after weaning. In addition, there was a substantial inhibition in adult feminine sexual behavior. Male rats that received either morphine or saline prenatally did not show any body weight differences, and most of the measures of masculine sexual behavior did not differ between the two groups. However, the male rats exposed to morphine had a significantly shorter post-ejaculatory intromission latency than the saline controls. Examination of cytosol estrogen receptor levels in the hypothalamus-preoptic area (HPOA) of both saline and morphine sulfate-treated female rats revealed essentially identical patterns of depletion and replenishment. Additionally, estrogen treatment was equally effective at inducing HPOA progestin receptor synthesis in both groups. These results show that prenatal morphine treatment at the times and dose level administered disrupts the development of reproductive function in females but has only minor effects on male reproductive function.

The purpose of this study was to investigate the effect of prenatal exposure to morphine sulfate on the development of reproductive function in female rats. Female rats exposed to morphine sulfate in utero (5-10 mg/kg on days 5-12 of gestation) exhibited varying dates of vaginal opening and a partial inhibition in adult feminine sexual behavior when compared to controls. However, the estrogen binding capacity of hypothalamic cytosols from morphine- and saline-treated females was identical. While we cannot rule out the possibility that the observed changes in reproductive function were an indirect result of morphine-induced alterations in maternal physiology or behavior, these results suggest that the development of feminine sexual behavior can be influenced during the early prenatal period and that morphine sulfate can affect the process of feminization.

The present study examined the actions of the antiprogestin RU 38486 on progesterone (P)-induced facilitation and sequential inhibition of estrous behavior and on brain cytosol progestin receptor (PR) levels in ovariectomized, estrogen-primed (0.5 micrograms of estradiol benzoate for 3 days) female rats. RU 38486 suppressed P-facilitated receptive and proceptive responses in a dose-dependent fashion when administered 1 hr prior to P. RU 38486 did not, however, block the sequential inhibitory effect of P. Indeed, when it was administered alone at a dose of 1 mg, animals were rendered behaviorally unresponsive to a P treatment 25 hr later. It is unclear whether RU 38486 is an agonist for P sequential inhibition of estrous behavior or if the apparent agonist action of RU 38486 actually results from a long-term antagonist effect. Estrogen-induced PRs remain elevated (35-55% above basal) in the hypothalamus (HYP) and preoptic area (POA) at 24 and 48 hr following the last estrogen injection. Thus P facilitation of estrous behavior is correlated with increased levels of cytosol PRs. RU 38486 reduced cytosol PRs in both brain regions to basal levels within 2 hr, and the levels remained suppressed 25 hr following the treatment. Hence there is a strong correlation between behavioral inhibition and suppressed cytosol PRs at both short (5 hr) and long (25 hr) intervals after RU 38486 administration. A P treatment which produced sequential inhibition of estrous responsiveness 24 hr later did not reduce the estrogen-induced level of cytosol PRs in either brain region. These results suggest that mechanisms in addition to induction of PRs may be necessary to ensure successful mating.

Both male and female rats vocalize ultrasonically during mating. Whereas male calls are known to facilitate female proceptive behavior, the female mating call has not been studied beyond spectrographic analysis. In this article, a series of experiments were done to examine the effects of the female's mating call on rat mating behavior. In the first experiment, females copulated with intact males before and after surgical devocalization. In the second experiment, intact females copulated twice with a male: once when they were able to hear and once when they were temporarily deafened with a medical ear mold. In the third experiment, tape recorded ultrasounds were placed in the presence of devocalized females while they were copulating with intact males. In the control condition, tape hiss was presented. In each experiment, it was observed that the female darted more if communication were disrupted (i.e., female devocalized or male deafened). Playback partially reduced darting to control levels. No other behavior was affected consistently across all experiments. Female calls might facilitate self-regulation of mating behavior, or they might focus the male's attention on her proceptive behaviors. It is also possible that the female calls could alter the stimulus properties of the male's behavior, indirectly affecting her own behavior.

Ejaculatory patterns were observed in normally reared, postpuberally castrated male and female rats treated with sex hormones and electrical shock in adulthood. In a preliminary experiment, 3 females treated with 50 microgram estradiol benzoate (EB) and 500 microgram progesterone (P) showed intromission and ejaculatory behavior when subjected to shock. Refractory periods were abnormally short and there was almost no postejaculatory vocalization. In Experiment 1, males and females were tested with electrical shock following daily treatment with 32 microgram EB with and without 500 microgram P on the test day. There was no difference between males and females in preejaculatory behavior, but females displayed abbreviated refractory periods and no postejaculatory vocalization. Progesterone had no observed effect. In Experiment 2 castrated males and females were subjected to shock after treatment with 8 microgram EB per day for 3 weeks. No P was given. Again females showed drastically reduced refractory periods and little vocalization. In Experiment 3, males and females treated with TP and shock displayed ejaculatory patterns, normal refractory periods and vocalization. Results show that female rats are capable of exhibiting the ejaculatory response without sex hormone treatment in perinatal life or androgen treatment in adulthood. It was also demonstrated that there is a sex difference in the postejaculatory behavior shown by estrogen-treated male and female rats.

Both male and female rats emit ultrasonic vocalizations during mating. In an earlier study, female vocalizations were shown to be associated with female darting: when the female was devocalized, she increased her rate of darting. In a preliminary playback study, tape-recorded female ultrasounds were presented to devocalized females paired with intact males. Darting was reduced towards presurgical control levels. In this study, the playback experiment was replicated using a refined procedure to present female ultrasounds to the mating rats. Taped vocalizations were obtained from females, and were presented in a manner which more closely approximated the patterning of ultrasound production seen in the intact female. Each rat pair was tested three times: once prior to the devocalization of the female, and twice afterwards. In one postsurgical test, taped ultrasounds were presented during mating; in the other, tape hiss was presented. Several patterns of behavior increased in frequency when the female was devocalized relative to the control condition, but decreased when tape recorded ultrasounds were presented. These patterns included female darting and approaches towards the partner by both males and females. The darting results confirm and extend the findings of our earlier paper. Changes in approach behavior were not observed previously because the male was confined in the earlier study. This study contains the first direct evidence that female vocalizations affect the socio-sexual behavior of the male.

The refractory period that characteristically follows ejaculation was abolished or significantly reduced by rostral midbrain lesions in male rats. The postejaculatory vocalization was also abolished or reduced, but other aspects of copulatory performance were unaffected. The results were attributed to disruption of biogenic amine pathways that pass from the ventral part of the rostral midbrain into the posterior hypothalamus.

The objective of the present study was to delineate the period of sensitivity to a single androgen exposure during the initial neonatal hours on the development of masculine and feminine copulatory behavior in female rats. Female rats were injected once with either 500, 50, or 5 micrograms testosterone propionate (TP) at either 1 or 24 hr after birth. Following castration in adulthood and TP replacement, the females were tested four times at weekly intervals in prolonged sessions for masculine copulatory behavior. One month following the masculine copulatory tests the females were tested for 3 weeks for feminine copulatory behavior with weekly increasing levels of estradiol benzoate (2.5, 10, and 25 micrograms) and progesterone (200 micrograms). The results demonstrate that a single injection of TP administered at either 1 or 24 hr after birth can significantly increase the capacity of female rats to exhibit ejaculation patterns and that the amount of androgen that is administered is critical in determining the levels of ejaculatory responding. Similarly, the females given high doses (50 and 500 micrograms) of TP at either 1 or 24 hr neonatally were almost completely defeminized. In contrast, however, the females treated with 5 micrograms TP at 1 and 24 hr showed different levels of lordotic performance indicating a greater sensitivity to androgen immediately after birth than at 24 hr in female rats as has been shown in male rats.

Prenatal exposure to a dopamine receptor blocking agent such as haloperidol (given to the mother at a dose of 0.5 mg/kg s.c. from day 4 to day 15 of gestation) produced subtle behavioural changes in rat pups. Haloperidol decreased the rate of ultrasonic vocalization in 4-day-old male pups removed from the nest. The changes in ultrasonic emission elicited by in utero exposure to this neuroleptic were markedly different from those produced by its administration during the early postnatal period. Moreover, adult male rats treated prenatally with haloperidol exhibited a significant increase in the intensity of ultrasonic 22 kHz post-ejaculatory calls emitted during sexual behaviour. The duration of the period of the 22 kHz calls emission was also significantly increased by haloperidol treatment. These results confirm that ultrasonic vocalization in rats is a sensitive indicator of subtle changes in adverse treatments administered during development.

The steroidal antiprogestin 17 beta-hydroxyl-11 beta-(4-dimethylaminophenyl)-17 alpha-(1-propynl)estra-4,9-dien-3-one (RU 38486) was administered systemically or was implanted into the ventromedial hypothalamus and other brain regions (habenula, preoptic area, interpeduncular region, in order to determine whether the compound could antagonize progesterone (P) activation of estrous responsiveness and whether the compound would exert its behavioral effects at the presumed site of P action and/or at other neural sites implicated in the regulation of female sexual behavior. RU 38486 (5 mg) administered sc 1 h before 200 micrograms P inhibited P facilitation of lordosis behavior in estrogen-primed rats. Intracerebral application of RU 38486 to the ventromedial hypothalamus reduced lordosis responses in 14 of the 25 animals tested. Similar implants in the habenula also inhibited lordosis in 5 of the 14 animals tested. Antiprogestin implants in the interpeduncular region and preoptic area were virtually without effect (1 of 7 inhibited in each group). Interactions of RU 38486 with steroid binding sites in the hypothalamus-preoptic area (HPOA) were also assessed. RU 38486 appeared to be a competitive inhibitor of progestin ([3H] R5020) binding in HPOA cytosols. Scatchard analysis of [3H]RU 38486 binding showed that when unlabeled P was used as the competitor to assess nonspecific binding, the antiprogestin bound with high affinity [dissociation constant Kd = 8.4 nm] to brain cytosols. In addition, the number of [3H]RU 38486 binding sites in HPOA cytosol increased by approximately 50% in estrogen-primed female rats. Competition studies indicated that unlabeled RU 38486 was the most effective competitor for [3H]RU 38486 binding but that P and R5020 were nearly as effective. Corticosterone, hydrocortisone, deoxycorticosterone, and triamcinolone also competed for [3H]RU 38486 binding but were somewhat less effective than the progestins. Testosterone and estradiol did not displace [3H]RU 38486 except at very high molar excesses. Thus RU 38486 appears to bind with highest affinity to HPOA progestin receptors, but it also binds to glucocorticoid receptors. These data are consistent with the interpretation that inhibition of estrous responsiveness by RU 38486 is associated with the antagonist's interference with brain progestin binding.

The putative neural target sites of progesterone (P) action for the facilitation of estrous behavior in female rats were reexamined using the hormone implant technique. Subjects were ovariectomized, estrogen-primed Long-Evans females. All were outfitted with bilateral double barreled cannulae assemblies consisting of 23-gauge guide cannulae and 28-gauge inserts. Subjects in Exp 1 received sc injections of individually determined threshold priming doses of estradiol benzoate. Estrogenic priming for subjects in Exp 2 was provided by sc placed Silastic capsules (5 mm) filled with 17 beta-estradiol-cholesterol (1:20). Each subject was tested for estrous behavior with a male after P-filled cannulae and after blank inserts were lowered into the brain. Behavioral tests were conducted 1 and 4.5 h postcannulae placement. Thirty-five of 40 females with P in the ventromedial hypothalamus exhibited high levels of lordosis behavior during the 4.5-h test. Moreover, 29 of the 35 exhibited solicitation behavior as well. Estrous behavior was not exhibited by these animals during the 1-h test. Implants in other regions of the brain (i.e. mesencephalic reticular formation, preoptic area, central grey, caudate putamen, and hippocampus; n = 80) did not consistently facilitate estrous responsiveness. In Exp 3, levels of estrous responsiveness were similar before and after adrenalectomy (n = 10); thus, facilitation of estrous behavior by P-filled implants in the ventromedial hypothalamus was not dependent upon activation of the hypothalamic-pituitary-adrenal axis.

Progesterone (P) action following estrogen priming is required normally for the facilitation of estrous behavior in female rats. Although mechanisms by which P exerts its influence on estrous responsiveness have not been elucidated, the primary site of action of P has been shown to be the ventromedial nucleus of the hypothalamus (VMN). The objective of the present series of experiments was to describe the temporal parameters of P action in the VMN in the facilitation of estrous behavior in estrogen-primed female rats. Subjects were female Long-Evans rats stereotaxically outfitted with 23-ga. guide cannulae directed towards the VMN. Crystalline P was applied directly to the brain tissue via bilateral 28-ga. insert cannulae, which could be inserted and removed easily through the guide cannulae. Animals were ovariectomized and estrogen primed with 5% estradiol Silastic capsules. They received a counterbalanced series of two experimental tests: one involving a manipulation with a P-filled implant, and another with a blank implant. In the first experiment, a significant increase in estrous responsiveness occurred only after 2 hr exposure of the VMN to P, whereas 4 hr were required for a full display of estrous behavior, including solicitation. In Experiment 2, P was lowered into the brain for either 1, 2, or 4 hr, and testing took place 4 hr after the lowering of the implant. It was found that 2 hr of P exposure was sufficient to facilitate full estrous responsiveness at 4 hr. In Experiment 3, it was revealed that the duration of estrous responsiveness was directly related to the time the P implant remained in the brain. In the fourth experiment, the time course of P retention in brain tissue, revealed by determination of 3H-progesterone levels in hypothalamus, agreed with the behavioral findings. Progesterone levels in the region of the VMN remained high while a P implant was in place, but declined rapidly after removal. A dual mechanistic hypothesis for P action in the facilitation of estrous behavior is presented.

In an attempt to obtain a more precise localization of the neural target site(s) of estrogen-priming action in the activation of estrous behavior, 30-gauge cannulae containing a preparation of 17 beta-estradiol diluted 1:250 with cholesterol were implanted bilaterally into the brains of ovariectomized rats. Subjects were tested for sexual behavior with intact males at 4-day intervals for 3 weeks beginning 2-3 days after stereotaxic surgery. Animals received a systemic injection of 0.5 mg progesterone 4-6 h before all but 1 test. After this treatment, estrous behavior was observed in 19 of the 20 animals judged to have both cannulae resting in or within 0.25 mm of the ventromedial nucleus of the hypothalamus (VMN). Virtually no estrous behavior was observed without the administration of progesterone. Behavioral scores decreased as a function of the distance of the cannulae tips from the VMN. Cholesterol implants in the same region (n = 8) were without effect. Implants in other regions of the brain (e.g. preoptic area, diagonal band of Broca, lateral habenula, amygdala, or cortex; n = 52) were ineffective. These results indicate that estrogenic stimulation of the region of the VMN alone is sufficient to prime the activation of estrous behavior in the ovariectomized rat.

We investigated the role of dopamine receptor subtypes in the regulation of ultrasonic vocalization and masculine copulatory behavior. Intact sexually experienced male Long-Evans rats were treated with saline, selective dopamine D1 (SKF 38393) and D2 (LY 171555) receptor agonists and with selective dopamine D1 (SCH 23390) and D2 (raclopride) receptor antagonists 15 and 30 min before the 30-min test session, respectively. Mating stimuli were ovariectomized female rats injected SC with estradiol benzoate (8 micrograms/0.1 ml/rat) and progesterone (200 micrograms/0.1 ml/rat), 48 and 4 hr before the test session, respectively. We found a decrease in the number of intromissions required to reach ejaculation in animals treated with SKF 38393 (10 mg/kg/IP), LY 171555 (doses ranging from 0.01 to 0.5 mg/kg/SC) and with raclopride (0.1 mg/kg/SC). LY 171555 reduced the postejaculatory vocalization (PEV) in a dose-dependent fashion with complete suppression at the highest dose. No other parameters of sexual behavior were affected by this treatment. Raclopride, a dopamine D2 receptor antagonist, antagonized the suppressive effects of the D2 agonist LY 171555 on the PEV (and also decreased the number of intromissions to reach ejaculation), whereas SCH 23390, a dopamine D1 receptor antagonist, did not. Raclopride, given alone at the dose of 0.5 mg/kg/SC, almost completely suppressed all behavioral activity, whereas the lower dose (0.1 mg/kg) decreased intromission frequency and increased the length of the 22 kHz PEV. Therefore, we suggest that 22 kHz PEV is under the control of dopamine D2 receptors.

The present study evaluated the facilitative actions of progesterone and the synthetic progestin R 5020 on estrous responsiveness in ovariectomized, estrogen-primed female rats. The dose-response and time-response characteristics of the behavior facilitating actions of both progesterone and R 5020 were measured. The threshold doses for the facilitation of estrous behavior in estrogen-primed female rats were 1 microgram of R 5020 and 100 micrograms of progesterone. These doses of progestins facilitated estrous responsiveness with a similar time course that approached maximum at one hour. To examine the possible mechanism(s) of action of each progestin the synthetic progestin antagonist RU 38486 was used. The inhibitory effects of RU 38486 on estrous behavior facilitated by a threshold dose of progesterone or R 5020 were found to be almost identical. RU 38486 (5 mg) administered 1 hr prior to progesterone or R 5020 suppressed lordosis behavior by 44% and 47% respectively. These results suggest that progesterone and R 5020 facilitate estrous responsiveness through the same mechanism.

Intraperitoneal administration of the opiate antagonist naloxone hydrochloride (30 mg/kg) to sexually experienced male rats caused a significant reduction in mount and intromission latencies, number of mounts preceding ejaculation and ejaculation latencies. Intraperitoneal adminstration of naloxone (30 mg/kg) also stimulated persistant non-copulators to begin mating and to ejaculate within a twenty minute test period. Conversely, intraperitoneal administration of morphine sulphate (6 mg/kg) as well as intraventricular injection of the endogenous opiate beta-endorphin (6 micrograms) produced a complete loss of copulatory behavior in male rats. The deficit in sexual behavior induced by beta-endorphin was correlated with a significant increase in hypothalamic norepinephrine levels. It is suggested that the endogenous opiates may be involved in the mediation of sexual behavior via an interaction with central catecholaminergic systems.

Sexual exhaustion of the male rat was studied with respect to several copulatory measures. The ejaculatory latency, intercopulatory interval, and intromission frequency demonstrate a U-shaped curve, exhibiting high values for the initial ejaculatory series, falling to a minimum at an intermediate ejaculatory series, and again increasing at exhaustion. The absolute refractory period of the postejaculatory interval (measured at vocalization termination) increases linerly, whereas the relative refractory period (the remaining portion of the postejaculatory interval) is a postively accelerating function. Partial recovery tests demonstrated that the preejaculatory measures and absolute refractory period have substantially returned to baseline values by Day 6, while the relative refractory period is still extended. The significance of these data to the theoretical modeling of sexual behavior is discussed.

The ventromedial nucleus of the hypothalamus (VMN) is thought to constitute an essential neural substrate for hormonal induction of female sexual behavior. In previous work, implants of dilute (0.4%) estradiol (E2) have been found to prime progesterone-facilitated female sexual behavior in rats only when they are within or very close to the VMN. In other studies, induction of cytosolic progestin receptors (PRc) in the hypothalamus has been correlated with systemic E2 priming of receptive behavior. We combined dilute E2 implants with a Palkovits punch microassay for PRc in the an effort to examine the relationship between estrous behavior and estrogen-induced PRc in the VMN of individual female rats. Ovariectomized rats were given bilateral guide cannulae aimed at the VMN and were primed for 3 days with 1) blank implants plus oil sc, 2) blank implants plus 0.5 micrograms E2 benzoate sc, 3) implants of 0.4% E2 in cholesterol, or 4) implants of 2.0% E2. On day 4 all rats received progesterone (P) sc and were tested for receptive behavior. The following week all subjects received the same estrogen-priming treatment but were killed on day 4 without P treatment. The VMN was punch dissected and assayed for PRc. High levels of lordosis were seen in the E2 benzoate-treated and 2.0% E2-implanted animals, while little behavior was seen in the oil-treated animals. Among 0.4% E2-implanted animals, higher levels of lordosis were observed in subjects with implant placements in the rostral than in the caudal VMN; however, lordosis quotients were not correlated with PRc induction measured in the entire VMN of individual animals. In general, our findings suggest that local estrogenic stimulation of target cells in the VMN is capable of both priming the estrous response and induction of P receptors. Whether these two effects are causally related awaits future study.

The effects of intracerebral estradiol benzoate (EB) implants upon lordosis behavior in castrated male rats were investigated. 27 guage EB-filled or Blank implants were placed unilaterally in either the preoptic area (POA) or the region of the ventromedial hypothalamic nucleus (VMH). When 0.5 microgram of EB was administered subcutaneously for eight days, males with EB implants had significantly higher lordosis quotients (LQ) than males with Blank implants. EB implants located in the region of the VMH were significantly more effective in stimulating lordosis responses than implants in the POA. These results indicate that, in male rats as in females, the VMH region is a primary site for the hormonal activation of lordosis behavior.