Enhanced D2 Agonism Induces Conditioned Appetitive Sexual Responses Toward Non-reproductive Conspecifics.

Brain mechanisms of sexual attraction toward reproductive partners develop from a systematic interrelationship between biology (nature) and learning (nurture). However, the causes of attraction toward non-reproductive partners are poorly understood. Here, we explored the role of Pavlovian learning under dopaminergic agonism on the development of sexual preference and brain activation for young male rats. During conditioning, adult sexually naïve males received either Saline (Saline-Paired) or the D2-receptor agonist quinpirole (QNP-Paired) and cohabited in contingency, or out of contingency (QNP-Unpaired) during 24 h with an almond-scented prepubertal juvenile male (PD25). Conditioning occurred every 4 days for three trials. Social and sexual responses were assessed four days after the last conditioning trial in a drug-free test, and males chose freely between a scented young male (PD37) and a novel receptive female. Four days later, males were exposed to the conditioned odor only and brain Fos-IR and serum testosterone were analyzed. Saline-Paired and QNP-Unpaired males displayed more non-contact erections (NCEs) and genital investigations for females, whereas QNP-Paired males expressed more NCEs and genital investigations for young males. In the QNP-Paired group, exposure to the young male-paired odor evoked more Fos-IR in limbic, hypothalamic and cortical areas, but no differences in serum testosterone were observed. Cohabitation with juvenile males during enhanced D2 agonism results in atypical appetitive sexual responses and a higher pattern of brain response for the young male-paired odor, with no changes in serum testosterone. We discuss the potential implications for the development of pedophilic disorder and perhaps other paraphilias.

Long-term effects of status epilepticus during infancy in male rats: Sexual behavior and brain response upon exposure to sexually receptive females.

Previous research in female rats showed that induction of status epilepticus (SE) during infancy impairs proceptive sexual behavior at the long run in adulthood but temporarily, since full proceptivity is recovered after four mating trials. In male rats, such equivalent effects have not been explored yet. Thus, SE was experimentally induced by injecting lithium chloride (3 mEq/kg, i.p.) in thirteen-day-old (P13) male pups and then, on P14, pilocarpine hydrochloride (100 mg/kg, s.c.). Controls received the same volume of saline. For Experiment 1, at P90, we analyzed c-Fos immunoreactivity (c-Fos-IR) as a measure of unconditioned brain activity after exposing them to sexually receptive females, but without physical contact. For Experiment 2, a different group of males was tested for locomotor activity, and their sexual behavior was assessed during five trials. Then, serum testosterone and corticosterone levels were measured. Our results showed that a lower proportion of SE males performed mounts, intromissions, and ejaculations, and repeated training did not improve their behavior. The levels of testosterone in SE males were reduced, but corticosterone, c-Fos-IR, and locomotion were similar to controls. These results suggest that SE during infancy impairs adult sexual behavior by reducing testosterone.

Brain activation associated to olfactory conditioned same-sex partner preference in male rats.

Sexual preferences can be strongly modified by Pavlovian learning. For instance, olfactory conditioned same-sex partner preference can occur when a sexually naïve male cohabits with an scented male during repeated periods under the effects of enhanced D2-type activity. Preference is observed days later via social and sexual behaviors. Herein we explored brain activity related to learned same-sex preference (Fos-Immunoreactivity, IR) following exposure to a conditioned odor paired with same-sex preference. During conditioning trials males received either saline or the D2-type receptor agonist quinpirole (QNP) and cohabitated during 24 h with a stimulus male that bore almond scent on the back as conditioned stimulus. This was repeated every 4 days, for a total of three trials. In a drug-free final test we assessed socio/sexual partner preference between the scented male and a receptive female. The results indicated that QNP-conditioned males developed a same-sex preference observed via contact, time spent, olfactory investigations, and non-contact erections. By contrast, saline-conditioned and intact (non-exposed to conditioning) males expressed an unconditioned preference for the female. Four days later the males were exposed to almond scent and their brains were processed for Fos-IR. Results indicated that the QNP-conditioned group expressed more Fos-IR in the nucleus accumbens (AcbSh), medial preoptic area (MPA), piriform cortex (Pir) and ventromedial nucleus of the hypothalamus (VMH) as compared to saline-conditioned. Intact males expressed the lowest Fos-IR in AcbSh and VMH, but the highest in MPA and Pir. We discuss the role of these areas in the learning process of same-sex partner preferences and olfactory discrimination.

Olfactory stimulation induces cerebellar vermis activation during sexual learning in male rats.

The cerebellum is a complex structure mainly recognized for its participation in motor activity and balance, and less understood for its role in olfactory processing. Herein, we assessed Fos immunoreactivity (Fos-IR) in the cerebellar vermis following exposure to different odors during sexual training in male rats. Males were allowed to copulate for either one, three or five sessions. One day after the corresponding session they were exposed during 60 min to woodshaving that was either: clean (Control), sprayed with almond scent (Alm) or from cages of sexually receptive females (RF). The vermis of the cerebellum was removed, cut in sagittal sections and analyzed for Fos-IR to infer activation. Our results showed that the cerebellum responded with more Fos-IR in the Alm and RF groups as compared to Control. More copulatory sessions resulted in more odor-induced Fos-IR, especially in the RF group. Accordingly, we discuss possible mechanisms on how the cerebellum mediates processing of both unconditioned and conditioned odors, and how sexual experience accelerates such process.

Effect of lithium-pilocarpine-induced status epilepticus on ultrasonic vocalizations in the infant rat pup.

Evidence shows that febrile convulsions induced in rat pups increase ultrasonic vocalizations (USVs); however, the effect of status epilepticus (SE) induced in developing rats on USVs has not been fully investigated. The goal of this study was to analyze USVs following lithium-pilocarpine-induced SE in fourteen-day-old (P14) rat pups. The rat pups were given 3-mEq/kg lithium chloride i.p. on the day before the induction of SE, which was carried out at P14 by subcutaneous injection of 100-mg/kg pilocarpine hydrochloride; control animals were given an equal volume of lithium chloride and saline on P13 and P14, respectively. Ultrasonic vocalizations were monitored at P15, P16, and P21 with a Mini 3 Bat Detector Ultra Sound Advice (15kHz-160kHz) set at 40±4kHz and digitally recorded in WAV format using the Audacity 1.3 beta software. A clear box (60×40×30cm) split down the middle with a holed wall was used; each pup was placed alone in one compartment, whereas its dam was placed on the other cage side at room temperature. Vocalizations were recorded over a 5-minute period, converted to sonograms and spectrograms, and analyzed using the Raven software. Parameters evaluated were as follows: USV frequency, latency to the first USV, and mean USV duration. There was a significant decrease in the latency (35.5±6.9s) and duration (50.8±8.6s) of USVs after SE compared with the control group (81.9±10.8s and 78.1±9.9s, respectively). Status epilepticus affected male and female rats differentially.

Involving the cerebellum in cocaine-induced memory: pattern of cFos expression in mice trained to acquire conditioned preference for cocaine.

Because of its primary role in drug-seeking, consumption and addictive behaviour, there is a growing interest in identifying the neural circuits and molecular mechanisms underlying the formation, maintenance and retrieval of drug-related memories. Human studies, which focused on neuronal systems that store and control drug-conditioned memories, have found cerebellar activations during the retrieval of drug-associated cue memory. However, at the pre-clinical level, almost no attention has been paid to a possible role of the cerebellum in drug-related memories. In the present study, we ought to fill this gap by aiming to investigate the pattern of neuronal activation (as revealed by cFos expression) in different regions of the prefrontal cortex and cerebellum of mice trained to develop conditioned preference for an olfactory stimulus (CS+) paired with cocaine. Our results indicate that CS+ preference was directly associated with cFos expression in cells at the apical region of the granule cell layer of the cerebellar vermis; this relationship being more prominent in some specific lobules. Conversely, cFos+ immunostaining in other cerebellar regions seems to be unrelated to CS+ preference but to other aspects of the conditioning procedure. At the prefrontal cortex, cFos expression seemed to be related to cocaine administration rather than to its ability to establish conditioned preference. The present results suggest that as it has been observed in some clinical studies, the cerebellum might be an important and largely overlooked part of the neural circuits involved in generating, maintaining and/or retrieving drug memories.

Cohabitation with receptive females under D2-type agonism in adulthood restores partner preference and brain dimorphism in the SDN-POA following neonatal gonadectomy in male rats.

Perinatal testosterone, or its metabolite estradiol, organize the brain toward a male phenotype. Male rodents with insufficient testosterone during this period fail to display sexual behavior and partner preference for receptive females in adulthood. However, cohabitation with non-reproductive conspecifics under the influence of a D2 agonist facilitates the expression of conditioned partner preference via Pavlovian learning in gonadally intact male rats. In the present experiment, three groups of neonatal PD1 males (N = 12/group) were either gonadectomized (GDX), sham-GDX, or left intact and evaluated for social preferences and sexual behaviors as adults. We then examined whether the effects of GDX could be reversed by conditioning the males via cohabitation with receptive females under the effects of the D2 agonist quinpirole (QNP) or saline, along with the size of some brain regions, such as the sexually dimorphic nucleus of the preoptic area (SDN-POA), suprachiasmatic nucleus (SCN), posterior dorsal medial amygdala (MeApd) and ventromedial hypothalamus (VMH). Results indicated that neonatal GDX resulted in the elimination of male-typical sexual behavior, an increase in same-sex social preference, and a reduction of the area of the SDN-POA. However, GDX-QNP males that underwent exposure to receptive females in adulthood increased their social preference for females and recovered the size in the SDN-POA. Although neonatal GDX impairs sexual behavior and disrupts partner preference and brain dimorphism in adult male rats, Pavlovian conditioning under enhanced D2 agonism ameliorates the effects on social preference and restores brain dimorphism in the SDN-POA without testosterone.

Pubertal stress in male rats: Effects on juvenile play behavior and adult sexual partner preference.

Puberty is a period of brain organization impacting the expression of social and sexual behaviors. Here, we assessed the effects of an acute pubertal stressor (immune challenge) on the expression of juvenile play (short-term) and sexual partner preference (long-term) in male rats. Juvenile play was assessed over ten trials at postnatal days (PND) (31-40) with age- and sex-matched conspecifics, and at PND35 males received a single injection of lipopolysaccharide (LPS, 1.5 mg/kg i.p.) or saline. Then, sexual partner preference was assessed at PND 60, 64, and 68, in a three-compartment chamber with a sexually receptive female and a male as potential partners simultaneously. The results confirmed that a single injection of LPS during puberty induced sickness signs indicative of an immune challenge. However, juvenile play was not affected by LPS treatment during the following days (PND36-40), nor was sexual behavior and partner preference for females in adulthood. These findings highlight that, while other studies have shown that LPS-induced immunological stress during puberty affects behavior and neuroendocrine responses, it does not affect juvenile play and sexual behavior in male rats. This suggests a remarkable resilience of these behavioral systems for adaptation to stressful experiences mediated by immune challenges during critical periods of development. These behaviors, however, might be affected by other types of stress.

The Anatomy, Histology, and Function of the Major Pelvic Ganglion.

This review provides a comprehensive analysis of the pelvic plexus and its regulation across various mammalian species, including rats, cats, dogs, and pigs. The pelvic and hypogastric nerves play crucial roles in regulating pelvic functions such as micturition, defecation, and erection. The anatomical organization of these nerves varies, forming either well-defined ganglia or complex plexuses. Despite these variations, the neurons within these structures are consistently regulated by key neurotransmitters, norepinephrine and acetylcholine. These neurons also possess receptors for testosterone and prolactin, particularly in rats, indicating the significant role of these hormones in neuronal function and development. Moreover, neuropeptides such as vasoactive intestinal peptide (VIP), substance P, neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL), and calcitonin gene-related peptide (CGRP) are co-released with neurotransmitters to modulate pelvic functions. This review highlights the complex interplay between neurotransmitters, neuropeptides, and hormones in regulating pelvic physiology and emphasizes the importance of hormonal regulation in maintaining the functionality and health of the pelvic plexus across different species.

The impact of subchronic hypercaloric and restriction diets on sexual behavior, serum testosterone, and prostate histology in rats.

The role of diet in health is crucial, with calorie intake playing a significant role. Hypercaloric diets (HD) often lead to adipose tissue accumulation and increased risk of chronic diseases, including reproductive impairments. By contrast, restriction diets (RD) help with weight loss, improve cardiovascular function, and ameliorate reproduction. Herein we sought to investigate the impact of subchronic HD and RD on body weight, sexual behavior, serum testosterone and prostate histology in rats. Hence, 10-week old male rats gained sexual experience during five trials with ovariectomized, hormone-primed females. Then at postnatal week PW15 the males were organized in three groups, depending on the feeding they received until PW18: HD, RD and standard diet (SD). During PW19-22 they were tested for sexual behavior, and at PW23 were euthanized for prostate histology (hematoxylin & eosin stain) and hormone analysis. Results indicated that HD males increased their body weight (16-23%) compared to SD and RD. Furthermore, HD males showed 65% less testosterone than RD males. The prostate of HD males revealed histological alterations, including a notable increase in epithelium height and other abnormal features, while no changes were observed in the performance of sexual behavior between HD and RD, although HD appeared to facilitate ejaculation when compared to SD. The histological features of RD males were comparable to SD males. Accordingly, we argue that subchronic modifications in calorie intake can alter body weight (in HD), serum testosterone levels (HD and RD in opposite directions), and prostate histology (in HD), while having no immediate effect on male sexual behavior.

Electrophysiological Characterization of Cerebellar Responses during Exploration and Grooming Behaviors in a Rat Model of Parkinsonism.

Parkinson's disease is currently a global public health challenge due to the rapid growth of aging populations. To understand its pathophysiology is necessary to study the functional correlation between the basal ganglia (BG) and the cerebellum, which are involved in motor control. Herein, we explored multiunit electrical activity (MUA) in the cerebellum of rats with induced Parkinsonism as a result of lesions following bilateral placement of electrodes and passing of current in the ventrolateral striatum (VLS). In one control group, the electrodes descended without electrical current, and another group was left intact in VLS. MUA was recorded in Sim B and Crus II lobes, and in the dentate nucleus (DN) during the execution of exploration behaviors (horizontal and vertical) and grooming. The lesioned and sham groups showed a decrease in MUA amplitude in the Crus II lobe compared to the intact group in all recorded behaviors. However, Sim B and DN did not express differences. Both electrical and physical insults to the VLS induced Parkinsonism, which results in less MUA in Crus II during the execution of motor behaviors. Thus, this type of Parkinsonism is associated with a decrease in the amplitude of Crus II.

Neurobiology of Maternal Behavior in Nonhuman Mammals: Acceptance, Recognition, Motivation, and Rejection.

Among the different species of mammals, the expression of maternal behavior varies considerably, although the end points of nurturance and protection are the same. Females may display passive or active responses of acceptance, recognition, rejection/fear, or motivation to care for the offspring. Each type of response may indicate different levels of neural activation. Different natural stimuli can trigger the expression of maternal and paternal behavior in both pregnant or virgin females and males, such as hormone priming during pregnancy, vagino-cervical stimulation during parturition, mating, exposure to pups, previous experience, or environmental enrichment. Herein, we discuss how the olfactory pathways and the interconnections of the medial preoptic area (mPOA) with structures such as nucleus accumbens, ventral tegmental area, amygdala, and bed nucleus of stria terminalis mediate maternal behavior. We also discuss how the triggering stimuli activate oxytocin, vasopressin, dopamine, galanin, and opioids in neurocircuitries that mediate acceptance, recognition, maternal motivation, and rejection/fear.

Multiunit recording of the cerebellar cortex, inferior olive, and fastigial nucleus during copulation in naive and sexually experienced male rats.

The sexual behavior of male rats constitutes a natural model to study learning of motor skills at the level of the central nervous system. We previously showed that sexual behavior increases Fos expression in granule cells at lobules 6 to 9 of the vermis cerebellum. Herein, we obtained multiunit recordings of lobules 6a and 7 during the training period of naive subjects, and during consecutive ejaculations of expert males. Recordings from both lobules and the inferior olive showed that the maximum amplitude of mount, intromission, and ejaculation signals were similar, but sexual behavior during training tests produced a decrease in the amplitude for mount and intromission signals. The fastigial nucleus showed an inverse mirror-like response. Thus, the cerebellum is involved in the neural basis of sexual behavior and the learning of appropriate behavioral displays during copulation, with a wiring that involves the cerebellar cortex, inferior olive, and fastigial nucleus.

Histological modifications of the rat prostate following transection of somatic and autonomic nerves.

It is known that hormones influence significantly the prostate tissue. However, we reported that mating induces an increase in androgen receptors, revealing a neural influence on the gland. These data suggested that somatic afferents (scrotal and genitofemoral nerves) and autonomic efferents (pelvic and hypogastric nerves) could regulate the structure of the prostate. Here we assessed the role of these nerves in maintaining the histology of the gland. Hence, afferent or efferent nerves of male rats were transected. Then, the ventral and dorsolateral regions of the prostate were processed for histology. Results showed that afferent transection affects prostate histology. The alveoli area decreased and increased in the ventral and dorsolateral prostate, respectively. The epithelial cell height increased in both regions. Efferent denervation produced dramatic changes in the prostate gland. The tissue lost its configuration, and the epithelium became scattered and almost vanished. Thus, afferent nerves are responsible for spinal processes pertaining to the trophic control of the prostate, activating its autonomic innervation. Hence, our data imply that innervation seems to be synergic with hormones for the healthy maintenance of the prostate. Thus, it is suggested that some prostate pathologies could be due to the failure of the autonomic neural pathways regulating the gland.

Quantification of neural and hormonal receptors at the prostate of long-term sexual behaving male rats after lesion of pelvic and hypogastric nerves.

Prostate function is regulated by androgens and a neural control via the pelvic and hypogastric nerves. As such, this sexual gland contains receptors for acetylcholine and noradrenaline, although it is unknown whether the expression of these receptors is affected by sexual behavior and even less by denervation of the gland. Thus, the purpose of this work was to evaluate the effect of repeated sexual behavior on the expression of noradrenaline, acetylcholine, and androgen receptors at the prostate, and how they are affected by denervation. To achieve this, we used sexually experienced males denervated at the pelvic or hypogastric nerves, or both. The messenger (mRNA) and protein for androgen, noradrenergic, and cholinergic receptors were evaluated. The weight of the gland and the levels of serum testosterone were also measured. We found that: (1) sexual behavior was not affected by denervation; (2) blood testosterone levels increased due to sexual behavior but such increase is prevented by denervation; (3) the weight of the ventral prostate increased with sexual behavior but was not affected by denervation; (4) AR messenger levels increased with sexual behavior but were not altered by denervation; (5) the messenger for noradrenergic and cholinergic receptors decreased after denervation, and those for muscarinic receptors increased, and (6) only AR protein decreased after denervation of both nerves, while those for other receptors remained unchanged. In summary, we show that the three receptors have different regulatory mechanisms, and that only androgen receptors are regulated by both autonomic systems.

Temporal coordination of pelvic and perineal striated muscle activity during micturition in female rabbits.

PURPOSE: To determine the motor basis of urine expulsion the activity of the pelvic (pubococcygeus) and perineal (bulbospongiosus and ischiocavernosus) muscles was recorded during micturition in anesthetized female rabbits. MATERIAL AND METHODS: Virgin female chinchilla rabbits were used for simultaneously recording cystometrograms and electromyograms of the pubococcygeus, ischiocavernosus and bulbospongiosus muscles. The particular contribution of each muscle during micturition was analyzed in another set of experiments in which each was inactivated by bilateral lidocaine injection. Bladder function was assessed using standard urodynamic parameters. RESULTS: Cystometrography showed that micturition comprises 2 phases, that is storage and voiding phases. During the latter phase no high frequency oscillations were recorded. On simultaneous electromyography recordings a temporal, coordinated activation of pelvic (pubococcygeus) and perineal (bulbospongiosus and ischiocavernosus) muscles was observed. During specific blockade of each muscle some modifications in urodynamic parameters were found. CONCLUSIONS: Our findings indicate a specific role for the pelvic and perineal muscles during feminine micturition.

Disruptive effects of neonatal gonadectomy on adult sexual partner preference and brain dimorphism in male rats: partial restoration with pubertal testosterone.

According to the organizational-activational hypothesis, testosterone or its metabolite estradiol, can organize the brain in a male direction (permanently or for long periods) if exposure occurs during a critical (sensitive) time of brain development like the prenatal period. Male rodents with insufficient levels of testosterone during such critical period would irreversibly fail to display sexual partner preference for receptive females in adulthood. However, exposure to testosterone during puberty is believed to function as a second critical period for organization of brain and behavior. Thus, in the present study we explored the effects of neonatal gonadectomy at postnatal day 1 (GNX) on the partner preference of adult males and the size of some sexually dimorphic regions in the brain like the SDN-MPOA, SCN, MeApd and VMH; and challenged its irreversibility by providing exogenous testosterone during puberty. Our results indicated that neonatal GNX impaired partner preference for females and reduced the size of SDN-MPOA, MeApd and VMH, but not SCN. GNX males restored with testosterone in PD30-PD59 (GNX + T) expressed partner preference for sexually receptive females and increased the size of SDN-MPOA and VMH, but not MeApd in adulthood. We conclude that neonatal castration and the lack of testosterone during the first month of life alters sexual behavior and brain dimorphism in adult male rats, but pubertal testosterone reverses the effects on behavior and brain dimorphism to some extent.

Fos expression at the cerebellum following non-contact arousal and mating behavior in male rats.

The cerebellum is considered a center underlying fine movements, cognition, memory and sexual responses. The latter feature led us to correlate sexual arousal and copulation in male rats with neural activity at the cerebellar cortex. Two behavioral paradigms were used in this investigation: the stimulation of males by distant receptive females (non-contact sexual stimulation), and the execution of up to three consecutive ejaculations. The vermis area of the cerebellum was removed following behavioral experiments, cut into sagittal sections, and analyzed with Fos immunohistochemistry to determine neuronal activation. At the mid-vermis region (sections from the midline to 0.1 mm laterally), non-contact stimulation significantly increased the activity of granule neurons. The number of activated cells increased in every lobule, but lobules 1 and 6 to 9 showed the greatest increment. In sexual behavior tests, males reaching one ejaculation had a high number of activated neurons similar to those counted after non-contact stimulation. However, two or three consecutive ejaculations showed a smaller number of Fos-ir cells. In contrast to the mid-vermis region, sections farthest from the midline (0.1 to 0.9 mm laterally) revealed that only lobule 7 expressed activated neurons. These data suggest that a well-delineated group of granule neurons have a sexual biphasic response at the cerebellar vermis, and that Fos in them is under an active degradation mechanism. Thus, they participate as a neural substrate for male rat sexual responses with an activation-deactivation process corresponding with the sensory stimulation and motor performance occurring during copulation.

Changes in multiunit activity pattern in cerebellar cortex associated to olfactory cues during sexual learning in male rats.

The cerebellum is a structure of the central nervous system which has been previously studied with different techniques and animal models and even humans, so it is associated with multiple functions such as cognition, memory, emotional processing, balance, control of movement, among others. Its relationship with sensory systems has already been explored, however, the role it plays in olfactory processing in the cerebellum is unclear. Several hypotheses have been proposed from work done in humans and animal models with neuroimaging and immunohistochemical techniques. Everything seems to indicate that the cerebellar function is of vital importance for the olfactory perception, being able to be controlling not only the olfactory aspect, but also the olfactory processing. In this study we analyzed the multiunit activity in the granular layer of the cerebellar vermis during olfactory stimulation: a session being sexually naive and during four sessions of sexual behavior learning. The amplitude was compared between male naive and sexual experts, as well as between olfactory stimuli. The amplitude of the sexually experienced rats showed the highest values compared to naive ones. Odor of receptive female causes the greatest amplitudes, however, in the control group the amplitude increased when they were sexually experts. The motor, sensory and associative learning generated by the acquisition of sexual experience modifies the activation pattern in the cerebellum by presenting neutral odors or associated with a reward.

A study of the prostate, androgens and sexual activity of male rats.

BACKGROUND: The prostate is a sexual gland that produces important substances for the potency of sperm to fertilize eggs within the female reproductive tract, and is under complex endocrine control. Taking advantage of the peculiar behavioral pattern of copulating male rats, we developed experimental paradigms to determine the influence of sexual behavior on the level of serum testosterone, prostate androgen receptors, and mRNA for androgen receptors in male rats displaying up to four consecutive ejaculations. METHODS: The effect of four consecutive ejaculations was investigated by determining levels of (i) testosterone in serum by solid phase RIA, (ii) androgen receptors at the ventral prostate with Western Blots, and (iii) androgen receptors-mRNA with RT-PCR. Data were analyzed with a one-way ANOVA followed by a post hoc application of Dunnett's test if required. RESULTS: The constant execution of sexual behavior did not produce any change in the weight of the ventral prostate. Serum testosterone increased after the second ejaculation, and remained elevated even after four ejaculations. The androgen receptor at the ventral prostate was higher after the first to third ejaculations, but returned suddenly to baseline levels after the fourth ejaculation. The level of mRNA increased after the first ejaculation, continued to increase after the second, and reached the highest peak after the third ejaculation; however, it returned suddenly to baseline levels after the fourth ejaculation. CONCLUSION: Four consecutive ejaculations by sexually experienced male rats had important effects on the physiological responses of the ventral prostate. Fast responses were induced as a result of sexual behavior that involved an increase and decrease in androgen receptors after one and four ejaculations, respectively. However, a progressive response was observed in the elevation of mRNA for androgen receptors, which also showed a fast decrease after four ejaculations. All of these changes with the prostate gland occurred in the presence of a sustained elevation of testosterone in the serum that started after two ejaculations. A consideration of these fast-induced changes suggests that the nerve supply plays a key role in prostate physiology during the sexual behavior of male rats.