Activation of mu opioid receptors in the medial preoptic area following copulation in male rats

The current study tested the hypothesis that sexual behavior is a biological stimulus for release of endogenous opioid peptides. In particular, activation of mu opioid receptors (MOR) in the medial preoptic area (MPOA), a key area for regulation of male sexual behavior, was studied in male rats. MOR endocytosis or internalization was used as a marker for ligand-induced receptor activation, utilizing confocal, electron, and bright microscopic analysis. Indeed, mating including one ejaculation induced receptor activation in the MPOA, demonstrated by increased immunoreactivity for MOR, increased numbers of endosome-like particles immunoreactive for MOR inside the cytoplasm of neurons, and increased percentage of neurons with three or more endosome-like particles inside the cytosol. Moreover, it was demonstrated that MOR activation occurred within 30 min following mating and was still evident after 6 h. Mating-induced internalization was prevented by treatment with the opioid receptor antagonist naloxone before mating, suggesting that mating-induced receptor activation is a result of action of endogenous MOR ligands. i.c.v. injections of MOR ligand [D-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin resulted in internalization of the MOR in a similar manner observed following mating. Finally, mating induced Fos expression in MOR containing neurons in the MPOA. However, naloxone pretreatment did not prevent Fos activation of MOR neurons, suggesting that Fos induction was not the result of MOR activation. In summary, these results provide further evidence that endogenous opioid peptides are released in the MPOA during male sexual behavior.     

Dopamine antagonists do not block conditioned place preference induced by paced mating behavior in female rats

The authors assessed the behavioral effects of dopamine (DA) receptor antagonists, Cis (Z) flupentixol and S(+)-raclopride L-tartrate, on conditioned place preference (CPP) induced by paced mating behavior. Ovariectomized female rats of the Wistar strain were used. The administration of amphetamine (1 mg/kg) induced a clear CPP that was completely blocked by the DA antagonists flupentixol (0.25 mg/kg) or raclopride (0.125 mg/kg). These doses had no effect on motor coordination. Female rats that mated in a pacing chamber developed a clear CPP. Neither flupentixol nor raclopride blocked the reward state induced by paced mating behavior. These results indicate that DA is not involved in the reward state induced by paced mating behavior in female rats.     

Sexual behavior induces naloxone-reversible hypoalgesia in male rats

The sensitivity to painful and sexual stimuli in male rats was markedly suppressed immediately after ejaculation and enhanced after the postejaculatory refractory period. The suppression of pain sensitivity induced by sexual activity was reversed, but sexual behavior was only slightly affected by intraperitoneal (i.p.) injection of the opioid antagonist naloxone (5 mg). Plasma concentrations of beta-endorphin-like immunoreactivity were unaffected by sexual activity. Injection of beta-endorphin (10 micrograms s.c.) markedly raised plasma concentrations of beta-endorphin-like immunoreactivity within 1 min of injection but did not affect the sensitivity to painful stimulation or the display of sexual behavior. It is suggested that while ejaculation may activate opioid receptor mechanisms, which affect the sensitivity to painful, but not sexual, stimuli, elevation of beta-endorphin in the blood does not affect the sensitivity to either sexual or painful stimuli in male rats.     

Perinatal inhibition of aromatization enhances the reward value of sex

Paced mating reduces the aversive properties and increases the positive characteristics of mating, inducing a reward state. Pacing is able to induce conditioned place preference (CPP), whereas nonpaced mating does not. The authors hypothesized that the aversive properties of mating are caused by androgens from adjacent males or from the mother during fetal life. To test whether aromatization of androgens induces the aversive properties of mating, female rats were treated perinatally with 1,4,6-androstatriene-3, 17-dione (ATD) to inhibit aromatization. When adults, these females were ovariectomized and hormonally primed to evaluate CPP after paced and nonpaced mating. During paced mating, control females showed higher return latencies after ejaculation, whereas ATD-treated females did not show a similar increase. In CPP tests, both paced and nonpaced mating induced a reward state in ATD-treated females, whereas only paced mating induced a reward state in control females. These results show that the perinatal inhibition of aromatization enhances the rewarding properties of mating, suggesting that estradiol induced the aversive properties of mating and/or modified the perinatal organization of the neuronal pathways in females.     

Activation of POMC neurons during general arousal but not sexual behavior in male rats

Exogenous opioids influence male rat sexual behavior, suggesting that endogenous opioid peptides are released during mating. Supporting this hypothesis, the authors recently showed that mating induced activation of mu opioid receptors. However, it is unknown which ligand(s) is acting on these receptors during mating. The current set of experiments tested the hypothesis that beta-endorphin-producing neurons, that is, proopiomelanocortin (POMC) neurons, are activated during sexual behavior. Mating-induced activation of POMC neurons was investigated during either the dark phase or the light phase, following different components of male rat sexual behavior or following control manipulations that resulted in general arousal. Results show activation of POMC neurons in the mediobasal hypothalamus following general arousal but not specifically related to sexual behavior per se. In addition, mating did not activate the subpopulation of POMC neurons that project to the medial preoptic nucleus. These results suggest that it is unlikely that POMC neurons contribute to the action of endogenous opioids in the brain area during sexual behavior but instead may contribute to the change in arousal state essential for the expression of sexual behavior.     

Ventromedial hypothalamic damage and sexual proceptivity in female rats

Sexual proceptivity is indicated by behaviors which reflect feminine initiation with respect to the occurrence of mating behavior. In this study we used ovariectomized female rats, and tested for sexual behavior in a relatively large arena with a sexually active male tethered in one corner. This testing situation gave the unrestrained female control over the occurrence of sexual interaction, and the frequency with which a female approaches a sexually active male provides a measure of sexual proceptivity. We find that administration of estrogen and progesterone to neurologically intact female rats induces sexual receptivity and increases the frequency with which they approach a sexually active male. Bilateral destruction of the ventromedial hypothalamus renders females non-receptive and virtually eliminates the tendency for females to approach males. Bilateral sagittal knife cuts which bracket the ventromedial region produce similar effects. Apparently, the ventromedial hypothalamus is importantly involved in the control of both receptive and proceptive components of feminine sexual behavior.     

Sex versus sweet: opposite effects of opioid drugs on the reward of sucrose and sexual pheromones

Endogenous opioids mediate some reward processes involving both natural (food, sweet taste) and artificial (morphine, heroin) rewards. In contrast, sexual behavior (which is also reinforcing) is generally inhibited by opioids. To establish the role of endogenous opioids for a newly described natural reinforcer, namely male sexual pheromones for female mice, we checked the effects of systemic injections of the general opioid antagonist naloxone (1-10 mg/kg) and the agonist fentanyl (0.1- 0.5 mg/kg) in a number of behavioral tests. Naloxone affected neither the innate preference for male-soiled bedding (vs. female-soiled bedding) in 2-choice tests nor the induction of place conditioning using male pheromones as rewarding stimuli, although it effectively blocked the preference for consuming a sucrose solution. In contrast, fentanyl inhibited the preference for male chemosignals without altering sucrose preference. These results suggest that, in macrosmatic animals such as rodents, opioidergic inhibition of sexual behavior might be due, at least partially, to an impaired processing of pheromonal cues and that the hedonic value of sweet-tasting solutions and sexual pheromones are under different opioid modulation.     

Endomorphin-1, effects on male sexual behavior

We examined the effect of endomorphin-1 (EM-1), an endogenous opioid peptide that binds selectively to the μ receptor, on male copulatory behavior in sexually vigorous Wistar rats. In the first experiment, four doses of EM-1 (1, 10, 50, and 100 μM) injected intracerebroventricularly produced a marked increase in ejaculation latency and interintromission interval and reduced the number of ejaculations during the test. In experiment 2 the effects of EM-1 were completely blocked by pretreatment with naloxone (5 mg/kg) 30 min prior to intracerebroventricular injection of EM-1 (100 μM). Collectively these results indicate that the activation of μ receptors by EM-1 modifies parameters associated with ejaculation (increases ejaculation latency and reduces the number of ejaculations) confirming that opioids are released during sexual behavior.     

Sex differences in opioid-mediated pain inhibitory mechanisms during the interphase in the formalin test

Many chronic pain conditions are more prevalent in women than men and both fundamental and clinical research supports the implication of endogenous pain inhibitory mechanisms. The goal of this study was to verify if sex differences on endogenous pain inhibitory mechanisms during the formalin test are opioidergic and modulated by sex hormones. Formalin tests were performed with naloxone hydrochloride, a non-selective opioid antagonist in intact and gonadectomized Sprague-Dawley rats of both sexes. Considering the sexual dimorphisms we found, where naloxone preferentially blocked the interphase in female rats, injections of all the possible combinations of mu- (naltrexone hydrochloride), delta- (naltrindole hydrochloride) and kappa-selective antagonists (norbinaltorphimine dihydrochloride) were given to evaluate the contribution of these opioid-receptor subtypes to the inhibitory mechanism during the interphase in intact females. Finally, the systemic administration of naloxone methiodide and intrathecal administration of naloxone hydrochloride in intact females allowed us to verify if the action of endogenous opioids that are liberated during the interphase takes place at the periphery or spinally, respectively. The results show that the interphase was almost completely inhibited by naloxone in females while it produced only a slight blockade in males. These results permitted us to conclude that opioids play a major role in the pain inhibitory mechanism of the interphase in females while a non-opioid mechanism seems to be responsible for this inhibitory pathway in males. Using gonadectomized animals of both sexes, we demonstrated the modulation of the opioidergic system of the interphase by sex hormones. The administration of different combinations of selective antagonists for mu-, kappa- and delta-opioid receptors in intact females permitted us to conclude that only the combination of kappa- and delta-selective antagonists significantly blocked the interphase. The same result was obtained with the combination of the three antagonists, confirming the results with systemic naloxone hydrochloride. Finally, intrathecal administration permitted us to support that the action of naloxone is primarily at the spinal level, even if a supraspinal action cannot be ruled out. These results are of particular interest in showing sexual dimorphisms in endogenous pain modulation mechanisms during the interphase of the formalin test. A clearer understanding of the difference between male and female endogenous pain inhibitory pathways should lead to a better understanding of the role of endogenous pain modulation deficits in certain chronic pain conditions.     

A peripheral, intracerebral, or intrathecal administration of an opioid receptor antagonist blocks illness-induced hyperalgesia in the rat

We used the tail-flick response of rats to study the role of opioid receptors in illness-induced hyperalgesia. An intraperitoneal injection of lithium chloride (LiCl) produced hyperalgesia that was blocked in a dose-dependent manner by subcutaneous injection of the opioid antagonist naloxone. Neither hyperalgesia nor its blockade by naloxone were due to variations in tail-skin temperature induced by LiCl. Hyperalgesia was also blocked when opioid receptor antagonism was restricted to (a) the periphery, by intraperitoneal administration of the quaternary opioid receptor antagonist naloxone methiodide; (b) the brain, by intracerebroventricular microinjection of naloxone; or (c) the spinal cord, by intrathecal microinjection of naloxone. These results document a pain facilitatory role of opioid receptors in both the peripheral and central nervous systems and are discussed with reference to their analgesic and motivational functions.     

"Some guys have all the luck": mate preference influences paced-mating behavior in female rats

In the present study, mating behavior was observed in female rats that were given the opportunity to mate with two males simultaneously. Three groups of females were tested: 1) sexually na?ve, naturally cycling rats in proestrous; 2) sexually na?ve, hormone-primed, ovariectomized (OVX) rats; and 3) hormone-primed, OVX rats tested 1 week after sexual receptivity testing. One male rat was determined to be the preferred male for each female, if she spent more time with him during a mating test. Independent of sexual experience, female rats were less likely to leave their preferred male than their non-preferred male following intromissions. However, when they left their preferred male, they returned to him faster than to their non-preferred male. This effect of preference was slightly more robust in the OVX rats. When female rats from Group 2 were tested with the same pair of males for 3 additional tests, each female's preference for a particular male was stable. That is, a female rat preferred the same male in approximately 3 out of the 4 tests, which is more likely than would be expected by chance. In a final experiment, pairs of male rats were tested with different females once weekly to determine if different females would prefer particular males consistently. Although no male rat was preferred by all females, females consistently preferred the same male from each pair during approximately 70% of the tests. In conclusion, female mate preference may have adaptive significance for the reproductive success of rats.     

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.     

Morphofunctional study of mammotropic cells following intraventricular administration of met-enkephalin

Summary An ultrastructural and morphometric study was carried out on the adenohypophyseal mammotropic cells of rats treated intraventricularly with an acute dose (150 g) of Met-enkephalin. In the female rats, clear features of cellular hyperactivity appeared after opioid administration. The changes affected the Golgi complex, the rough endoplasmic reticulum, the mature and immature secretory granules and the images of exocytosis. Such changes did not appear when naloxone was administered before the opioid, and naloxone induced an increase in the numerical density of lysosomal dense bodies with lipoid inclusions. In the male animals, administration of an identical dose of Metenkephalin caused only a few significant changes, similar to those observed in the controls. It is concluded that Metenkephalin administered intraventricularly causes evident modifications in the mammotropic cells of female rats whereas such changes in the male animals are not significant.     

An NMDA antagonist in the MPOA impairs copulation and stimulus sensitization in male rats

Systemic injections of an NMDA antagonist have been shown to impair mating in male rats. One site where glutamate and its NMDA receptors may contribute to mating is the medial preoptic area (MPOA), which is vital for male sexual behavior. Glutamate is released in the MPOA during copulation, and especially at the time of ejaculation. We report here that the NMDA antagonist MK-801, microinjected into the MPOA, impaired copulatory behavior in sexually na?ve as well as experienced males. In rats tested both as na?ve and after sexual experience, drug treatment produced more profound impairment in na?ve males. In addition, MK-801, microinjected into the MPOA before each of 7 noncopulatory exposures to receptive female rats, resulted in copulatory impairments on a drug-free test on Day 8, relative to aCSF-treated rats; their behavior was similar to that of males that had not been preexposed to females. Therefore, NMDA receptors in the MPOA contribute to the control of copulation and stimulus sensitization. Glutamate, acting via NMDA receptors, regulates many neural functions, including neuronal plasticity. This is the first demonstration that a similar mechanism in the MPOA sensitizes male rats to the stimuli from a receptive female, and thereby enhances their behavior.     

Opioid control of the fetal stretch response: implications for the first suckling episode.

Upon their first experience with milk, fetal rats express a stretch response that is similar to the postnatal behavior exhibited by infant rats at the nipple. Fetuses also possess a functional opioid system that is activated by prenatal milk exposure. The opioid receptor antagonist naloxone and the specific kappa antagonist nor-binaltorphimine blocked the stretch response and prevented the increase in rearlimb activity that is typically induced by milk. The mu antagonist beta-funaltrexamine blocked the stretch while permitting the expression of rearlimb activity. The kappa agonist U50,488 promoted rearlimb activity in the absence of milk, whereas the mu agonist [D-Ala2,NMe-Phe4,Gly5-ol]-enkephalin (DAMGO) exerted little influence on fetal behavior. Fetuses pretreated with U50,488 stretched to nonmilk stimuli (saline or lemon), but fetuses pretreated with DAMGO did not. Opioid activation is part of a chain of events that culminates in the fetal stretch response and may be important in promoting milk ingestion during the newborn's first suckling episode.     

"Nice guys finish last": influence of mate choice on reproductive success in Long-Evans rats

The present study was designed to determine if male physiology and male reproductive behavior predict reproductive success in Long–Evans rats. Mating behavior was observed in sexually naïve, naturally cycling female rats during behavioral estrous that were given the opportunity to mate with two males simultaneously. DNA analysis of offspring born following these mating encounters was used to identify the paternity of each pup. In order to assess the effect of mate choice during these mating encounters on reproductive success, one male rat in each pair was categorized as the preferred mate if the female spent more time (> 50%) with him during the mating test of the present study. Furthermore, each male in the pairs was categorized as “attractive” or “non-attractive” by computing the number of females that preferred each male across many mating tests. Similar to results reported in Lovell et al. (2007), during 76% of these mating tests the same male rat in each pair was preferred by different female rats. Overall attractiveness of individual male rats predicted reproductive success in the present study. Interestingly, “attractive” males sired significantly FEWER pups than “non-attractive” males. Neither behavioral (e.g., latency to first sexual stimulation, number of sexual stimulations) nor physiological measures (e.g., body weight, urinary testosterone levels) of male rats predicted their reproductive success. In conclusion, the present results indicate that certain features of some males are more attractive to females, but attractive males are at a reproductive disadvantage (as measured by the number of pups sired). Although basal urinary testosterone levels did not differ between males that sired the majority of pups in a litter and males that sired few or none of the pups in a litter, aggression and/or other physiological measures of fertility (e.g., penile reflexes) may differ between males that are attractive to females and those that have a reproductive advantage.     

Oxytocin mediates the antidepressant effects of mating behavior in male mice

A significant association between plasma oxytocin (OT) levels and depression has been demonstrated. A recent study found that sexual activity and mating with a female induced the release of OT in the central nervous system of male rats. Here we examined the effect of mating behavior on depression-related behavior in wild-type (WT) and OT receptor-deficient (OTR KO) male mice. The WT males showed a reduction in depression-related behavior after mating behavior, but the OTR KO mice did not. Application of an OTR antagonist inhibited mating behavior-induced antidepressant effect in WT males. OT may mediate the antidepressant effects of mating behavior.     

Penile deafferentation and the effect of mating experience on sexual motivation in adult male rats

In the present study naive male rats were used, in which before or after housing for 5 days with receptive females penile deafferentation was performed by means of transection of the pudendal nerves. In subsequent mating tests the males were only able to exhibit mounting behavior. It appeared that the animals with mating experience prior to penile denervation, mounted significantly more than the animals that had been denervated before housing with the females. Furthermore the last group showed longer contact latencies, which were similar to those observed in denervated animals that had not been housed with females. It is concluded that the reinforcing value of copulatory performances upon sexual motivation in the rat is completely dependent upon the integrity of penile afferent innervation.     

Effects of prenatal morphine exposure on rat heterotypical sexual behavior.

Prenatal exposure to morphine inhibits ovarian steroid-dependent lordosis behavior in female rats, and enhances certain components of male sexual behavior in male rats. In the present study, the effects of mid to late gestational morphine exposure on male sexual behavior in females and on female sexual behavior in males were examined in adult offspring. Gonadectomized male rats were injected at weekly intervals with 30 or 60 microg estradiol benzoate and 1.0 mg progesterone and tested for female sexual behavior with stimulus males on 2 consecutive weekly tests. Ovariohysterectomized (OVX) females were injected with 500 microg testosterone propionate (TP) daily for 15 days and tested for male sexual behavior with stimulus females on the last day of TP injection and 1 week later, after TP withdrawal. Prenatal morphine exposure increased the expression of male sexual behaviors in female rats, but it did not increase lordosis behavior in male rats. Thus, exposure to morphine during gestation alters male and female sexual behavior in young adult animals. Because prenatal morphine exposure both defeminized and masculinized adult sexual behavior in female rats, it is possible that female brain development is more vulnerable to prenatal insult such as opiate exposure.