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Arginine vasotocin facilitation of advertisement calling and call phonotaxis in bullfrogs.

Although the neuropeptide arginine vasotocin (AVT) is found in several auditory and vocalization regions of the bullfrog (Rana catesbeiana) brain, its functions in these areas are unknown. We examined the effects of AVT injection on two auditory-evoked reproductive behaviors: advertisement calling in male bullfrogs and call phonotaxis in female bullfrogs. AVT (500 micrograms; intraperitoneal) significantly increased advertisement call frequency and decreased calling latency in male bullfrogs, compared to saline injection. This dose of AVT also significantly decreased the time required for female bullfrogs to reach a call source as well as the latency of females to leave the starting position during call playback. In both males and females, these effects were significant at 0.5 hr and persisted until at least 2 hr after injection. AVT thus facilitated display of two sexually-dimorphic reproductive behaviors in bullfrogs. These effects may be due to direct effects of AVT on auditory processing regions in the bullfrog brain.

Animals↗

Maternal age as a factor in determining the reproductive and behavioral outcome of rats prenatally exposed to ethanol.

Nulliparous Long-Evans rats were bred at one of four different ages and assigned to one of three treatment groups within each age condition. Maternal ages were 9, 18, 32, and 36 weeks. Treatment groups were ethanol (E), administered by gavage as 8 g/kg in two divided doses on days 10-14 of gestation, pair-fed (PF) controls, administered as an isocaloric sucrose solution by gavage on days 10-14 of gestation, and ad lib fed controls (C). All offspring were surrogate fostered shortly after delivery to untreated recently parturient dams. Litter sizes were standardized to 8 on the day of birth. Offspring were assessed longitudinally for growth, mortality, and behavior (olfaction, locomotor activity, maze learning, avoidance acquisition and startle). Approximately 85% of the 36 week old dams did not produce viable litters. In the remaining maternal age conditions, ethanol delayed offspring olfactory orientation and increased locomotor activity, the latter dissipating after 50-60 days of age. These ethanol-related effects occurred independent of maternal age condition. Maternal age, independent of ethanol, was a factor which reduced litter size and offspring weight up to 50 days, but produced few effects on behavior. The combination of maternal age and prenatal ethanol interacted to increase pregnancy loss (oldest maternal age), reduce offspring weight up to day 99 (oldest and middle maternal age), alter olfactory orientation performance (oldest and middle maternal age), reverse the typical ethanol-induced increase in activity for males in the figure-8 test (oldest maternal age group), shift the pattern of open-field activity, and change errors in a complex water maze. Not all of these interactions turned out to be specific to the ethanol X old maternal age condition. Several of the interactions occurred in both the old and middle maternal age conditions. The only effect of old maternal age that interacted strongly with ethanol was in their combined effects on reproduction. Here the combination of the two factors increased maternal mortality, the number of early pregnancy losses, and the number of litters where all members were dead or resorbed. It was concluded that short-term prenatal ethanol combined with advanced maternal age produces additive interactions on pregnancy success without affecting longer-term outcomes, while young maternal age showed no clear detrimental effects compared to the middle maternal age reference group.

Aging↗

Acute fasting decreases sexual receptivity and neural estrogen receptor-alpha in female rats.

Acute food deprivation or chronic food restriction suppresses reproduction in female mammals. Although a link between undernutrition and ovarian function is well established in rats, a similar link with reproductive behavior in this species is yet to be described. Therefore, we compared the display of estrous behaviors induced by exogenous steroid hormone treatment in ovariectomized fed and fasted rats. In addition, estrogen receptor-alpha immunoreactivity (ERIR) was measured in fed and fasted animals to determine whether changes in behavior were associated with changes in the number of detectable ERIR-containing cells in several brain regions. Fasting for 74 h decreased lordosis quotients (LQ) and lordosis ratings (LR) in ovariectomized, steroid-primed rats. The number of detectable ERIR cells decreased after a 74-h fast in the mid-region of the arcuate (ARC), paraventricular (PVN) and ventromedial nuclei of the hypothalamus (VMH) and the ventral bed nucleus of the stria terminalis (BST) but did not change in a number of other areas examined. Taken together, these data demonstrate that, similar to the effect on the reproductive-endocrine axis, food deprivation for 74 h suppresses steroid-induced display of lordosis in adult, female rats. Furthermore, this suppression in sexual receptivity is associated with a decrease in ERIR in a number of areas, including the VMH, a region of the hypothalamus known to be critical for the display of reproductive behaviors in female rats.

Animals↗

Oxytocin and female sexuality.

A search of the literature has been prepared to determine how oxytocin may affect sexual and reproductive in women. Many animal studies suggest that oxytocin induces a variety of reproductive behaviors, including grooming, sexual arousal, orgasm, gamete transport, nesting, birthing, and specific maternal behaviors such as breast-feeding and bonding between mother and infant. These actions are apparently facilitated by the 'priming' effect on certain cells by sex and steroid hormones - as the brief case report would also suggest. However, no adequate double-blind trial has confirmed the observations from this report in women. Only animal studies have been performed, albeit over a wide range of species. A variety of other causes and effects of sexual interest and arousal relating to oxytocin are considered, including some of those in males. More research is needed to clarify the role of oxytocin in human reproductive behaviors, including its potential 'aphrodisiac' or prosexual effect in women in the presence of the sex-steroid hormones.

Female↗

Differential fertility as a mechanism maintaining balanced polymorphisms in Sardinia.

Women's fertility, gathered from the 1961 Italian population census, and estimates of heterozygote frequencies for thalassemia and G6PD deficiency (Siniscalco et al. 1961, 1966) in 52 Sardinian villages were examined to study at the population level the mechanisms that have maintained the stability of these polymorphisms over long periods. Sardinian villages were classified according to low or high frequency of heterozygotes, and the reproductive behavior of the women living in these areas was analyzed. A high mean number of children per woman and a low percentage of women without children with a high heterozygote frequency was demonstrated. The observed differential fertility and sterility were interpreted as being the result of different numeric ratios within each area between normal homozygous and heterozygous women, who were less and more resistant, respectively, to malarial infection, according to Haldane's theory. The effect of differing degrees of malaria on fertility rates has been demonstrated previously (Zei et al. 1990). To account for the effect of the genetic and epidemiological composition of an area on reproductive behavior, we classified data on women's fertility and sterility by heterozygote frequency level and malarial morbidity level. A combined and direct effect of inherited and acquired immunities on fertility and sterility rates was shown. The level of endemicity in an area may contribute to decreasing or increasing fitness, which is already influenced by the stable balanced polymorphisms.

Adult↗

Endogenous opioid-immunoreactive neurons of the ventromedial hypothalamic nucleus concentrate estrogen in male and female rats.

Estrogen stimulates expression of proenkephalin mRNA in neurons of the hypothalamic ventromedial nucleus, and evidence is accumulating that synaptic release of one of the peptide end products, met-enkephalin, influences events that regulate reproductive behavior. To address the question of whether estrogen acts directly on neurons that synthesize met-enkephalin or indirectly through a separate neuronal population, we combined estrogen autoradiography with endogenous opioid peptide (EOP) immunohistochemistry. In agreement with previous studies, the ventrolateral subdivision of the hypothalamic ventromedial nucleus was densely packed with EOP-immunoreactive cells. In males, 48% of the estrogen-concentrating cells of the ventrolateral subdivision of the hypothalamic ventromedial nucleus contained EOP, and, in females, 27% of the estrogen-concentrating cells contained EOP. These findings indicate that estrogen acts directly on neurons that express EOP and suggest a mechanism that underlies sexually differentiated reproductive behavior.

Animals↗

The influence of environment, sex, and innate timing mechanisms on body temperature patterns of free-ranging black-tailed prairie dogs (Cynomys ludovicianus).

Mechanisms that influence body temperature patterns in black-tailed prairie dogs are not well understood. Previous research on both free-ranging and laboratory populations of black-tailed prairie dogs (Cynomys ludovicianus) has suggested that reductions in ambient temperature and food and water deprivation are the primary factors that stimulate torpor in this species. In other species, however, torpor has been shown to be influenced by a multitude of factors, including innate circadian and circannual timing mechanisms, energy status, and reproductive behaviors. Our objective was to clarify the influence of weather, sex, and intrinsic timing mechanisms on the body temperature patterns of free-ranging black-tailed prairie dogs. We monitored body temperatures of eight adult (>1 yr) prairie dogs from November 1999 to June 2000. Prairie dogs showed distinct daily and seasonal body temperature patterns, which reflected changes in ambient temperatures that occurred during these periods. These patterns of daily and seasonal heterothermy suggest that body temperature patterns of black-tailed prairie dogs may be driven by an innate timing mechanism. All prairie dogs entered torpor intermittently throughout winter and spring. Torpor bouts appeared to be influenced by precipitation and reductions in ambient temperature. Our results also suggest that reproductive behaviors and circadian timing may influence torpor in this species.

Animals↗

Alteration by estrogen of the nucleoli in nerve cells of the rat hypothalamus.

Estrogen is accumulated from the blood by nerve cells in the ventromedial nucleus of the hypothalamus and can facilitate female reproductive behavior by acting on this region of the brain. This cell group was examined in ovariectomized female rats, given estrogen or control treatment, by use of light and electron microscopy. A significantly greater portion of the nerve cells in the estrogen-treated animals had protuberances on their nucleolar surfaces, apparent under the light microscope. The fine structure of such protuberances included dense, aggregated material, which is shown to contain DNA by the sodium tungstate staining technique. Because increased numbers of such protuberances were found in nuclei of cells of the experimental group where previous studies demonstrated a significant increase in ultrastructural signs of biosynthetic activity, they may be associated with increased RNA synthesis. Thus, they could indicate, ultrastructurally, increased synthetic rates for RNA in nerve cells through which estrogen promotes reproductive behavior.

Animals↗

Gonadal steroid hormone modulation of nociception, morphine antinociception and reproductive indices in male and female rats.

The purpose of this study was to examine how gonadal steroid hormones modulate basal nociception and morphine antinociception relative to regulating reproduction in the adult rat. Male and female Sprague-Dawley rats were either gonadectomized (GDX) or sham-gonadectomized (sham); GDX males were implanted subcutaneously with capsules containing testosterone (T), estradiol (E2), dihydrotestosterone (DHT), E2 and DHT, or nothing (0). GDX females received E2, T, or empty (0) capsules immediately after surgery, and vehicle or progesterone (P4) injections at 4-day intervals. Basal nociception and morphine antinociception were tested 28 days after surgery on 50 degrees C and 54 degrees C hotplate tests, and reproductive behavior and physiology were assessed shortly thereafter. There were no significant differences in baseline hotplate latencies among the male treatment groups, but morphine was significantly more potent in sham and GDX+T males than in GDX+0 males. The ability of T to increase morphine's potency was approximated by its major metabolites E2 and DHT, given together but not alone. Baseline hotplate latencies were higher in sham females tested during diestrus than in those tested during estrus. Morphine was significantly more potent in sham females tested during proestrus and diestrus than in those tested during estrus. Baseline hotplate latencies were significantly higher, and morphine was significantly less potent in GDX+E2, GDX+E2/P4 and GDX+T females than in GDX+0 females. All group differences in basal nociception and morphine antinociception observed on the 50 degrees C hotplate test were smaller and generally non-significant on the 54 degrees C hotplate test. Steroid manipulations produced the expected changes in reproductive behaviors and steroid-sensitive organs. These results demonstrate that in adult rats, gonadal steroid manipulations, that are physiologically relevant, modulate (1) basal nociception in females but not males, and (2) morphine's antinociceptive potency in both males and females.

Animals↗

Sperm competition: defining the rules of engagement.

Genetic and cell biological analyses of sperm behavior in the female reproductive tract are providing important clues to the mechanisms of sperm competition, a form of sexual selection that is an important force that shapes reproductive behavior, physiology and morphology in a wide range of species.

Animals↗

Modification of gonadotropin releasing hormone (GnRH) mRNA expression in the retinal-recipient Thalamus.

Although the environmental cues that trigger reproductive behaviors are known for many species, the mechanisms through which these signals influence the neurochemistry of the brain to produce behavior have been elusive. In this study, we describe a retinally modulated system of gonadotropin releasing hormone (GnRH) producing neurons in the thalamus of the plainfin midshipman fish, Porichthys notatus. Previously, we cloned and sequenced the cDNA for prepro-GnRH in midshipman. Here, using in situ hybridization, we localized prepro-GnRH mRNA to the ventrolateral nucleus of the thalamus, three divisions of the preoptic area, the ganglion of the terminal nerve, and the olfactory bulb. Since the thalamus, terminal nerve ganglion, and preoptic area have been associated with visual functions, we investigated the retinal connections in midshipman. In particular, biocytin tract tracing delineated a reciprocal connection between the ventrolateral nucleus of the thalamus and the retina. Retinofugal projections are exclusively contralateral. Experimental manipulation of this retinalthalamic loop through complete optic nerve transection shows that GnRH mRNA expression in the contralateral ventrolateral nucleus may be influenced by the retina. We hypothesize that a reciprocal retinothalamic GnRH circuit is important in modulating the expression of seasonal reproductive behaviors.

Animals↗

Sexual dimorphism in the vasotocin system of the bullfrog (Rana catesbeiana).

Arginine vasotocin (AVT) is widespread in amphibian brains, where its levels have been correlated with reproductive behaviors. To better understand which neural systems are involved in central actions of AVT, we used immunocytochemistry to compare the distribution of AVT in the brains of male and female bullfrogs (Rana catesbeiana). AVT-immunoreactive cells were observed in the septal nucleus, amygdala pars lateralis, magnocellular preoptic area, suprachiasmatic nucleus, and hypothalamus. AVT-immunoreactive cells were also found in the pretrigeminal nucleus, but only in animals killed in the fall. Immunoreactive fibers were broadly distributed in hypothalamic and extrahypothalamic areas. The most obvious sex differences were found in the amygdala pars lateralis, where the density of immunoreactive cells and fibers was significantly greater in male than in female bullfrogs. In addition, in the habenular nucleus, males had a denser distribution of AVT-immunoreactive fibers than females. In the suprachiasmatic nucleus, AVT-immunoreactive cells were larger in females than in males but did not differ in number. Since the areas that showed sex differences in AVT distribution have also been implicated in control of reproductive behaviors, they may form the neural substrates for the effects of AVT on sexually dimorphic behaviors in amphibians.

Animals↗

Altered endocrine and behavioral responses with reproductive aging in the male Japanese quail.

Experiments were conducted to further characterize the age-related decline in reproductive capability previously described in the male Japanese quail. Behavioral testing of a large number of males in several age groups confirmed earlier results that showed a significant reduction in courtship and mating behavior by 80 weeks of age. Intact males, 78 weeks of age, that showed no reproductive behavior were given Silastic implants containing testosterone. This treatment restored behavior in 80% of the males. Gonadal regression induced by exposure to shortened photoperiod followed by transfer to a stimulatory photoperiod did not successfully restore the endocrine or behavioral components of reproduction. Males of two age groups, 42 and 208 weeks of age, were categorized as behaviorally active or inactive and then castrated. Eight weeks later they were given implants containing testosterone. Older males required longer exposure time to the exogenous steroid and in some cases, an additional implant to restore mating behavior as compared to younger males. The presence or absence of behavioral activity prior to castration did not affect the length of time required for restoration of behavior. Challenge with exogenous luteinizing hormone-releasing hormone revealed that there was reduced release of luteinizing hormone in aged, behaviorally inactive males as compared to young males. Results of these experiments in addition to previous data give evidence that age-related changes occur at the level of the hypothalamus and pituitary as well as at the level of the gonads.

Aging↗

Evidence for estrogen receptor in cell nuclei and axon terminals within the lateral habenula of the rat: regulation during pregnancy.

The habenular complex is involved in several estrogen-dependent reproductive behaviors in female rats, namely, sexual behavior, maternal behavior, and postpartum sexual behavior. Although it is known that estrogen acts in other brain regions to mediate these behaviors, it is not known whether estrogen may also act directly on the habenular complex. To address this possibility, we examined this region for the presence of estrogen receptor (ER). This analysis was carried out in separate experiments by using in situ hybridization, immunocytochemistry at the light and electron microscopic levels, and steroid autoradiography. Neurons within the lateral habenula (LHb), but not the medial habenula, express ER mRNA, contain ER immunoreactivity (ER-ir) in their nuclei, and concentrate radiolabelled estradiol, providing strong evidence for the presence of functional ER in the lateral habenula. There were also ER-ir containing punctate fibers within the LHb, which, at the electron microscopic level, in part, proved to be axons and presynaptic axonal terminals. Both the level of ER-ir in cell nuclei and the density of ER-ir fibers within the LHb were regulated during the course of pregnancy and the postpartum period, suggesting that the sensitivity of the LHb to estrogen may be altered during this time. Taken together, these results demonstrate that the LHb is likely a more estrogen-sensitive region than was previously considered, and they suggest alternative mechanisms of action for ER. ER within the LHb may play a critical role in the involvement of the LHb in estrogen-dependent female reproductive behaviors.

Animals↗

Preproenkephalin mRNA levels are regulated by acute stress and estrogen stimulation.

Enkephalins facilitate female reproductive behavior. Within the limbic system and hypothalamus, estrogen induced the expression of preproenkephalin (PPE) mRNA. Estrogen injection caused a biphasic increase in the PPE mRNA levels within the ventromedial hypothalamic nucleus and posterodorsal medial amygdala. The first peak of PPE mRNA levels occurred within an hour, and the second 24 to 48 h after subcutaneous injection of estrogen. The present studies indicated that the rapid first peak of PPE mRNA expression was stress induced, whereas the second peak was estrogen induced. In the posterodorsal medial amygdala but not in the ventromedial hypothalamic nucleus, the antiestrogen, tamoxifen, did not inhibit the first peak, but blocked the second peak of PPE mRNA expression. Subcutaneous oil injection induced a 1-h peak of PPE mRNA levels but not a 24-h peak. Peak levels of plasma corticosterone were coincident with peak PPE mRNA levels. Adrenalectomy plus a constant, low level of corticosterone eliminated the injection-induced increase of corticosterone levels and the subsequent increase in PPE mRNA expression in the ventromedial hypothalamic nucleus and posterodorsal medial amygdala. The present results indicate that both stress steroids and estrogen positively regulate PPE mRNA levels in the ventromedial hypothalamic nucleus and posterodorsal medial amygdala. These results are consistent with the hypothesis that acute, mild stress may contribute to the activation of circuits that facilitate reproductive behavior in the female.

Administration, Oral↗

Estrous cycle variations in levels of cholecystokinin immunoreactivity within cells of three interconnected sexually dimorphic forebrain nuclei. Evidence for a regulatory role for estrogen.

The central part of the medial preoptic nucleus (MPNc), the encapsulated part of the bed nucleus of the stria terminalis (BSTe), and the posterodorsal part of the medial nucleus of the amygdala (MeAp) are all though to be involved in the neural control of female reproductive behavior, as well as other neuroendocrine mechanisms. Although the developmental importance of gonadal steroids during the perinatal period on these sexual dimorphisms is well known, an understanding of possible activational effects on these cell groups of circulating gonadal steroids in the adult is less clear. In the present study we evaluated the number of cholecystokinin (CCK)-immunoreactive cells present within MPNc, BSTe, and MeAp of regularly cycling female rats over the estrous cycle. In addition, the effects of ovariectomy and estrogen replacement on CCK staining were also examined. The number of CCK-immunoreactive cells within each cell group varied over the estrous cycle with the fewest cells present in animals sacrificed while in diestrus. Proestrous female rats showed a greater number of cells within each nucleus, while intermediate numbers were found for animals in estrus. These changes appear to be due, at least in part, to changes in levels of circulating estrogen, since subcutaneous implants of estradiol prevented the decline in the number of CCK-stained cells within MPNC, BSTe, and MeAp that was seen in untreated, ovariectomized female rats. Thus, the present findings support the hypothesis that levels of CCK within cells of these three sexually dimorphic cell groups are regulated by circulating gonadal steroids within a physiologically relevant time frame and may possibly contribute to the activation of female reproductive behavior as well as other neuroendocrine functions.

Amygdala↗

The foam production system of the male Japanese quail: characterization of structure and function.

The research described here characterizes a unique neuromuscular system involved in reproductive behavior--the foam production system of the male Japanese quail (Coturnix japonica). Male quail produce a large amount of foam that is transferred to the female during copulation, enhancing male fertilization success. The source is the foam gland complex, a large sexually dimorphic, androgen sensitive, external protuberance of the dorsal cloaca, consisting of glandular units interdigitated with striated muscle fibers of the sphincter cloacae muscle (mSC). Electromyographic (EMG) analysis of mSC activity in freely moving males interacting with females revealed different characteristics of the EMG signal during copulation, voiding of excreta, and other mSC movement. The amount of mSC activity and also the amount of foam produced were greatly increased by the presence of a female behind a screen. Denervation of mSC eliminated normal mSC movement and also abolished foam production, confirming that mSC activity is the mechanism for foam production. The spinal cord locations of the motoneurons innervating the major cloacal muscles, including mSC, were determined by injecting cholera-toxin conjugated horseradish peroxidase into each muscle. Labelled somata with multiple primary dendrites were located in Area IX of the lateral motor column of synsacral segments 7, 8, or 9 or 8, 9, and 10. The motoneurons serving mSC were intermingled with those projecting to the other cloacal muscles, but there were differences in the rostralcaudal placement of these neural populations. Thus mSC activity is an integral part of the male's reproductive behavior, mSC activity can be socially stimulated, and mSC activity occurring in anticipation of copulation is likely to be functionally significant. Continued investigation of this highly accessible system has the potential to shed light on the mechanisms by which complex motor acts are produced and hormonally regulated.

Animals↗

Distribution analysis of the two chicken estrogen receptor-alpha isoforms and their transcripts in the hypothalamus and anterior pituitary gland.

Estrogen plays a key role in the control of reproductive behavior and in the regulation of the neuroendocrine system. To elucidate the mechanisms by which it controls these functions it is important to understand how estrogenic effects are mediated. We have investigated the distribution of the two isoforms of the chicken estrogen receptor alpha (cER-alpha) protein; the previously characterized cER-alpha 66 and a new N-terminal truncated isoform, cER-alpha 61. Immunolocalization demonstrated the presence of cER-alpha 66 protein in hypothalamic areas, principally the nucleus septalis lateralis, bed nucleus striae terminalis medialis, nucleus preopticus medialis, and nucleus infundibuli hypothalami, and in the anterior pituitary gland. When the distribution of ER-alpha immunoreactive cells was compared using the antibodies H 222 (directed against the hormone-binding domain) and ER 221 (directed against the 21-amino acid N-terminus), no apparent differences could be detected. Because this immunocytochemical approach was not able to distinguish whether full-length cER-alpha 66 is the only isoform observed in the ER-positive regions or whether both cER-alpha receptor isoforms are present, SI nuclease assays were performed to compare the relative abundance in these regions of the two distinct classes of cER-alpha mRNA variants (A1-D and A2), which encode the cER-alpha 66 and cER-alpha 61 protein isoforms, respectively. In cockerels and hens, both variants of cER-alpha mRNA are expressed in the anterior pituitary gland and basal hypothalamus with a dominance of the mRNA that encodes cER-alpha 66, whereas the mRNA that encodes cER-alpha 61 was not detectable in the anterior hypothalamus. Therefore, because both receptor isoforms differ in their ability to modulate estrogen target gene expression in a promoter and cell type-specific manner, these differences may mediate the pleiotropic actions of estrogen in reproductive behavior and neuroendocrine functions.

Animals↗