The role of delta-opioid receptors in estrogen facilitation of lordosis behavior

The present study investigated the role of delta-opioid receptors (ORs) in estrogen facilitation of female rat reproductive behavior (lordosis). Infusion of 2 microg of the selective delta-OR agonist [D-Pen(2),D-Pen(5)]-enkephalin (DPDPE), into the third ventricle facilitated lordosis behavior in ovariectomized (OVX) rats injected with estrogen (E) 48 and 24 h before behavioral testing. Pretreatment with the selective delta-OR antagonist naltrindole (NTDL) blocked DPDPE effects on lordosis behavior. Ventricular infusion of NTDL (40 microg) also suppressed lordosis behavior in fully receptive OVX rats primed with both E and progesterone (P). In addition, NTDL blocked lordosis behavior when infused into the ventromedial nucleus of the hypothalamus (VMH) but not into the medial preoptic area (mPOA). Site-specific infusion of DPDPE into the VMH had dose-dependent, dual effects on lordosis behavior. While a very low dose of DPDPE (0.01 microg) facilitated lordosis behavior, a higher dose (1.0 microg) inhibited receptivity in OVX rats primed with E and a low dose (50 microg) of P. We used 3H-DPDPE to measure the density of delta-ORs in OVX rats treated with vehicle or with E by receptor autoradiography. E treatment did not have any effect on the density of DPDPE binding sites in the VMH, mPOA, medial amygdala, or caudate putamen. The behavioral effects of the ligands used in this study suggest that activation of delta-OR in the VMH by endogenous opioids facilitates estrogen-dependent lordosis behavior.     

Projections of the sexually dimorphic calcitonin gene-related peptide neurons of the preoptic area determined by retrograde tracing in the female rat

The medial preoptic area of the rat exhibits morphologic sex differences and is implicated in the control of sexually dimorphic behavior and function. Neurons expressing calcitonin gene-related peptide (CGRP) within the anteroventral periventricular (AVPV) and medial preoptic nucleus (MPN) of the medial preoptic area exhibit female-dominant sex differences in number through organizational and activational effects of gonadal steroids. The present study used retrograde tracing experiments to establish the projections of the AVPV and MPN CGRP neurons in the female rat. After the intraperitoneal administration of Fluoro-Gold to female rats (n = 5), we were unable to detect retrograde tracer in any CGRP-immunoreactive cells of the hypothalamus. Intracerebral injections of 50- to 100-nl volumes of Fluoro-Gold into the mediobasal hypothalamus resulted in up to 70% of CGRP neurons in the AVPV and MPN containing retrograde tracer. Similar large volume tracer depositions in the lateral septum, periaqueductal gray, two likely CGRP projection sites, resulted in no labeling of preoptic CGRP neurons. Experiments using small volume (30-nl) injections of Fluoro-Gold and green fluorescent microspheres at multiple sites in the mediobasal hypothalamus (n = 18) revealed that approximately 60% of AVPV and 30% of MPN neurons expressing CGRP were projecting to the region of the tuberal and ventral premammillary nuclei, with a minor projection to the dorsomedial nucleus. These findings demonstrate a major projection of the preoptic CGRP neurons to the posterior hypothalamus in the female rat and support further a functional role for these neurons in the sexually dimorphic regulation of reproductive functioning.     

Estrogen-receptor immunoreactivity in hamster brain: preoptic area, hypothalamus and amygdala

The distribution of estrogen-receptor containing cells in the preoptic area, hypothalamus and amygdala of female Syrian hamster brain was studied by immunocytochemical methods. Dense populations of estrogen-receptor immunoreactive (ER-IR) cells were found in the medial preoptic area, the bed nucleus of the stria terminalis, amygdala, ventral and lateral parts of the hypothalamus, and the arcuate nucleus. Injection of estradiol caused a decrease in estrogen-receptor immunoreactivity (ERIR) containing cells within one hour, a decrease that may reflect a change in the ability of the occupied estrogen receptor to bind the particular antibody (H222) used rather than down-regulation of the estrogen receptor. Our findings on the distribution of estrogen-receptor containing cells in these areas using an immunocytochemical technique are consistent with and extend the findings of others using autoradiographic and in vitro binding techniques to study estrogen receptor distribution in hamster brain.     

Autoradiographic differentiation of mu, delta, and kappa opioid receptors in the rat forebrain and midbrain

While there is an abundance of pharmacological and biochemical evidence to suggest the existence of multiple opioid receptors, their precise localization within the brain is unclear. To help clarify this issue, the present study examined the distributions of the mu, delta, and kappa opioid receptor subtypes in the rat forebrain and midbrain using in vitro autoradiography. Mu and delta receptors were labeled with the selective ligands 3H-DAGO (Tyr- D-Ala-Gly-MePhe-Gly-ol), and 3H-DPDPE (D-Pen2, D-Pen5-enkephalin), respectively, while the kappa receptors were labeled with 3H-(-)bremazocine in the presence of unlabeled DAGO and DPDPE. Based on previous findings in our laboratory, the labeling conditions were such that each ligand selectively occupied approximately 75% of each of the opioid sites. The results demonstrated that all 3 opioid receptor subtypes were differentially distributed in the rat brain. Mu binding was dense in anterior cingulate cortex, neocortex, amygdala, hippocampus, ventral dentate gyrus, presubiculum, nucleus accumbens, caudate putamen, thalamus, habenula, interpeduncular nucleus, pars compacta of the substantia nigra, superior and inferior colliculi, and raphe nuclei. In contrast, delta binding was restricted to only a few brain areas, including anterior cingulate cortex, neocortex, amygdala, olfactory tubercle, nucleus accumbens, and caudate putamen. Kappa binding, while not as widespread as observed with mu binding, was densely distributed in the amygdala, olfactory tubercle, nucleus accumbens, caudate putamen, medial preoptic area, hypothalamus, median eminence, periventricular thalamus, and interpeduncular nucleus. While all 3 opioid receptor subtypes could sometimes be localized within the same brain area, their precise distribution within the region often varied widely. For example, in the caudate putamen, mu binding had a patchy distribution, while delta and kappa sites were diffusely distributed, with delta sites being particularly dense ventrolaterally and kappa sites being concentrated ventromedially. These results support the existence of at least 3 distinct opioid receptors with possibly separate functional roles.     

Biochemical and Immunocytochemical Assessment of Neural Progestin Receptors Following Estradiol Treatments that Eliminate the Sex Difference in Progesterone-Facilitated Lordosis in Guinea-Pigs

A single injection of estradiol benzoate (10 μg), while highly effective in ovariectomized female guinea-pigs, does not prime castrated males to display progesterone-facilitated lordosis. In contrast, adult males and females exhibit the same, high degree of progesterone-facilitated lordosis when primed with two, 2.0 μg injections of free estradiol-17ß (pulse regimen). We compared neural progestin receptor induction after these different estradiol treatments by in vitro radioligand binding assays and immunocyto-chemistry. Binding assays confirmed previous observations of lower concentrations of cytosol progestin receptors in the mediobasal hypothalamus-preoptic area in estradiol benzoate-treated males than in females. No such sex difference was observed in animals that had been exposed to the behaviorally effective estradiol pulse regimen; rather, hypothalamic-preoptic area progestin receptor concentrations in these animals did not differ from the low levels observed in males treated with the behaviorally ineffective estradiol benzoate regimen. Immunocytochemical analysis revealed progestin receptor-immunoreactivity in fewer cells in the medial preoptic nucleus-anterior hypothalamic nucleus, periventricular preoptic area and arcuate nucleus of estradiol pulse- as compared to estradiol benzoate-treated males and females. Estradiol benzoate treatment induced progestin receptor-immunoreactivity in more cells in the medial preoptic area and ventrolateral hypothalamus than estradiol pulses in males, but not in females. Surprisingly, in these regions estradiol benzoate-treated males had significantly more progestin receptor-immunoreactive cells than females. These experiments yield two major findings: First, as has been shown in rats, the display of progesterone-facilitated lordosis is not inflexibly differentiated according to sex in guinea-pigs. Furthermore, reduced concentrations of estradiol-induced progestin receptors in the hypothalamus and preoptic area cannot account for the lack of progesterone-facilitated lordosis that is observed following priming with estradiol benzoate in males. Secondly, in the medial preoptic area and ventrolateral hypothalamus of female guinea-pigs, estradiol pulses are as effective as estradiol benzoate in inducing progestin receptors. These observations, taken together with the finding of equal behavioral efficacy of the two estradiol treatments, are consistent with the hypothesis that estradiol-induced progestin receptors in these regions of the brain are involved in progesterone-facilitated lordosis in female guinea-pigs.     

Hypothalamic Estrogen Receptor-lmmunoreactivity in Prepubertal vs Adult Female Guinea Pigs

Juvenile guinea pigs (18–20 days old) rarely display lordosis in response to estradiol and progesterone treatments that elicit sexual behavior in adult females. Nor do immature animals release a preovulatory-like surge of luteinizing hormone in response to estradiol. In vitro radioligand binding assays have revealed similar concentrations of estrogen receptors in thehypothalamus and preoptic area of prepubertal and adult guinea pigs. The aim of the present study was to compare estrogen receptor-immunoreactivity in a variety of forebrain regions of immature and adult guinea pigs, to determine whether age differences in estrogen receptor levels inmore discrete portions of the hypothalamus and preoptic area exist. Forebrain tissue from juvenile (17 days) and adult females (> 6 weeks), ovariectomized 6 days previously, was processed forestrogen receptor-immunoreactivity, using Abbott Laboratories' H222 anti-human estrogen receptor antibody. Juveniles had estrogen receptor-immunoreactive cells in all of the same regions as adults: medial preoptic area, medial preoptic nucleus, bed nucleus of the stria terminalis, periventricular, paraventricular, dorsomedial and arcuate nuclei, ventrolateral and anterior hypothalamic regions, and amygdala. Among the areas in which estrogen receptor-immunoreactivity was quantified (medial preoptic area, medial preoptic nucleus, anterior periventricular nucleus, arcuate nucleus and ventrolateral hypothalamus), the only region in which an age difference in estrogen receptor-immunostaining was observed was the rostral portion of the ventrolateral hypothalamus. Juvenile females had, on average, 30% fewer estrogen receptor-immunoreactive cells in asample of this region than adults (440 ± 25 vs 626 ± 25, P = 0.001). These data are consistent with the hypothesis that insufficient estrogen receptors in the rostral ventrolateral hypothalamus may underlie, in part, the markedly deficient responses of juvenile female guinea pigs to estradiol.     

Regulation of galanin receptor GalR1 mRNA expression by ovarian steroids in oestrogen receptor alpha-immunoreactive neurones: identification of distinct populations of neurones in the preoptic area

This study examined whether gonadal steroids are involved in regulating galanin receptor 1 (GalR1) mRNA expression in neurones that contain oestrogen receptor alpha (ERalpha), in three regions of the preoptic area (POA) known to be involved in the control of gonadotropin secretion. Double-labelling immunohistochemistry using an antibody against the ERalpha and in situ hybridization experiments using a 35S-labelled riboprobe specific for GalR1 mRNAs revealed that ERalpha is expressed in a large proportion of GalR1 mRNA-expressing neurones of the POA in the ovariectomized (OVX) female rat. Oestradiol (E2) and oestradiol plus progesterone (E2 + P) treatments of OVX rats significantly decreased the proportion of GalR1 mRNA/ERalpha immunoreactive (ERalpha-IR) neurones in the anteroventral periventricular nucleus (AVPV), medial preoptic nucleus (MPN) and medial preoptic area (MPO). The expression of GalR1 mRNA in ERalpha-IR neurones varied according the hormonal status of the female animals. In the AVPV, during the oestrous cycle, the hybridization signal significantly increased at oestrus. E2 and E2 + P treatments of OVX rats did not induced significant variation of levels of GalR1 mRNAs in ERalpha-IR neurones. In the MPN, E2 treatment of OVX rats resulted in significant increase in GalR1 mRNA expression in ERalpha-IR neurones. Similarly, levels of the GalR1 hybridization signal increased during afternoon of proestrus and oestrus. In the MPO, treatment of OVX rats with E2 + P significantly decreased GalR1 mRNA expression in ERalpha-IR neurones. The expression of GalR1 mRNA did not change during the oestrous cycle in this area. These findings suggest that the hypothalamic action of galanin on gonadotopin-releasing hormone (GnRH) secretion may pass through the specific population of GalR1/ERalpha-IR neurones of the MPN in mediating the oestrogen action on the GnRH system at the moment of the luteinizing hormone surge.     

Evidence that Orexin-Containing Neurones Provide Direct Input to Gonadotropin-Releasing Hormone Neurones in the Ovine Hypothalamus

Orexins A and B (ORX) have been added recently to the growing list of neuropeptides implicated in feeding and drinking behaviour as well as neuroendocrine function. In the present study, we have used single and dual labelling immunohistochemistry and a rabbit polyclonal anti-orexin-A antibody, which recognizes both ORX A and B, to examine ORX pathways in the sheep hypothalamus. ORX immunoreactive cells were distributed in the dorsomedial hypothalamic nucleus, lateral hypothalamic area, zona incerta and perifornical area; a few cells were also observed in the anterior hypothalamic area. In contrast to distribution in the rat brain, most of the ORX immunoreactive cells are localized to the dorsomedial hypothalamic nucleus and perifornical area; scattered cells are found in lateral hypothalamic area. ORX immunoreactive fibres were widely distributed throughout the hypothalamus and preoptic area with dense innervation of the medial preoptic area and bed nucleus of stria terminalis. Dual labelling demonstrated widespread expression of the long form of the leptin receptor within all ORX cells that were examined. Thirty percent of the gonadotropin releasing hormone (GnRH) cells that were examined had ORX immunoreactive terminals in close contact with no regional or sex differences. FluoroGold injections into the preoptic area retrogradely labelled a subpopulation of ORX cells in the lateral hypothalamic/perifornical area, showing ORX cells of this region project to the preoptic and could potentially provide input to GnRH cells. These findings suggest an integral role for ORX in the regulation of GnRH cells in the sheep and thus provide evidence of a novel mechanism whereby leptin can influence reproductive neuroendocrine function.     

Effect of Vorozole, an Aromatase Enzyme Inhibitor, on Sexual Behavior, Aromatase Activity and Neural Immunoreactivity

Aromatase enzyme is essential for the expression of normal sexual behavior in many mammals and birds. Here we report that vorozole (R83842), a non-steroidal aromatase inhibitor, blocks sexual behavior in the female musk shrew. In addition, vorozole treatment lowers aromatase activity in male and female preoptic area, and reduces plasma estradiol concentrations in females. Our findings confirm and extend results demonstrated in other species, conducted with the active enantiomer (R83842), or the racemic mixture (R76713, racemic vorozole). We also report that vorozole treatment affects the immunocytochemical distribution of aromatase immunoreactivity (AROM-ir) in musk shrew brain. The histological identification of neurons that contain this enzyme has been difficult in mammals. Several aromatase enzyme antisera have been developed and used in brain, and each gives a different pattern of immunoreactivity. Moreover, despite the fact that aromatase activity is very high in the bed nucleus of the stria terminalis, several amygdala nuclei, the preoptic area and hypothalamus, AROM-ir in these regions has been very limited. The distribution of AROM-ir in female musk shrew brain tissues is modified by treatment with vorozole prior to sacrifice. Female musk shrew brains contain aromatase immunoreactive cell bodies, as reported previously, in the central amygdala, lateral septum and to a limited extent in the bed nucleus of the stria terminalis (BST). Brains of females treated with vorozole show additional immunoreactivity in the preoptic area, hypothalamus, and medial amygdala, and have a broad distribution of AROM-ir in several subdivisions of the BST. Several sexual dimorphisms are apparent in musk shrews brains after treatment with vorozole. We have quantified this sexual dimorphism in the medial preoptic area (MPO) by counting immunoreactive cells. In both the rostral and caudal portions of the MPO, female brains contain significantly fewer AROM-ir cell bodies than males. These data are in complete agreement with sex differences in biochemical analyses of aromatase activity in the MPO. At this time we do not know if these dimorphisms are the result of differences in circulating levels of steroids in males and females, and/or if the AROM-ir nuclei regulate sexually dimorphic behaviors.     

The organization of neural inputs to the medial preoptic nucleus of the rat

There is general agreement that the medial preoptic nucleus (MPN) receives projections from widespread regions of the brain, although there are significant discrepancies in the literature with regard to certain specific inputs. Therefore, we have reexamined the inputs to this nucleus with both retrograde and anterograde axonal transport techniques. First, injections of the retrograde tracers true blue, SITS, or wheat germ agglutinin were made into the region of the MPN and the distribution of retrogradely labeled cells was charted. Then, autoradiographic material was used to confirm the results of the retrograde studies, to identify the route taken by fibers projecting to the MPN, and to describe the distribution of projections with respect to the three cytoarchitectonic subdivisions of the nucleus. The results indicate that the MPN receives inputs from widely distributed areas in both the forebrain and brainstem, and that these inputs appear to be distributed topographically within the three cytoarchitectonic subdivisions of the nucleus. Direct inputs to the MPN arise from all major areas of the hypothalamus (except for the median and magnocellular preoptic nuclei, the supraoptic and suprachiasmatic nuclei, and the medial and lateral mammillary nuclei). Projections from nuclei within the periventricular zone of the hypothalamus end primarily in the medial part of the MPN, while inputs from the lateral zone are mainly confined to the lateral part of the nucleus, as are projections from the nuclei within the medial zone, except for those from the anterior and ventromedial nuclei, which appear to be more widespread. The MPN receives major inputs from limbic regions including the amygdala, ventral subiculum, and ventral lateral septal nucleus, all of which end preferentially in the lateral part of the MPN. In contrast, the projection from the encapsulated part of the bed nucleus of the stria terminalis appears to end preferentially in the central part of the MPN and in immediately adjacent regions of the medial subdivision. In addition, the MPN may receive relatively sparse inputs from infralimbic and insular cortical areas, the nucleus accumbens, and the substantia innominata. Finally, ascending serotoninergic projections from the raphe nuclei appear to terminate principally in the lateral part of the MPN, whereas inputs from regions containing noradrenergic cell groups are chiefly distributed to the central and medial parts of the nucleus. Other brainstem regions that appear to provide modest inputs include the ventral tegmental area, central tegmental field, periaqueductal gray, pedunculopontine nucleus, and the peripeduncular nucleus.(ABSTRACT TRUNCATED AT 400 WORDS)     

Intrahypothalamic Mu-, not Delta- or Kappa-Opioid Receptor Activation Causes Growth Hormone Secretion

The possible effects of opioid receptor agonists on growth hormone (GH)-releasing factor or somatostatin neurons were examined by measuring the effects of localized intracerebral injections of mu-, delta- and kappa-selective agonists on GH secretion. Serial GH concentrations were measured in plasma in unanaesthetized male rats chronically prepared with venous and intracerebral cannulae, before and after treatment with bilateral intracerebral injections of opioid agonists in the preoptic anterior hypothalamic area and medial basal hypothalamus. In the medial basal hypothalamus, injections of the mu-agonist DAGO (Tyr-D-Ala-Gly-(Me)Phe-Gly-ol) caused dose-responsive increases in GH, the maximally effective dose being 0.001 nmoles. Injection of 10,000-fold higher doses of the delta-agonist DPDPE ([D-Pen, D-Pen]enkephalin) and the kappa-agonist U50,488H were also effective in stimulating GH secretion. In the preoptic anterior hypothalamic area, DAGO caused dose-responsive increases in GH, the maximally effective dose being 0.01 nmoles. U50.488H was ineffective at 1,000-fold higher doses while DPDPE was effective at 100- to 1,000-fold higher doses. We conclude that hypothalamic mu-opioid receptor activation on or near somatostatin or GH-releasing factor neurons causes GH secretion. Opioids capable of acting on other opioid receptors may also stimulate GH secretion, though only at doses that seem likely to affect mu-receptors.     

Presence of sex difference of cytochrome P-450 in the rat preoptic area and hypothalamus with reference to coexistence with oxytocin

Localization of female type cytochrome P-450 (F1) in the preoptic area and hypothalamus of the rat was examined immunocytochemically using antiserum against purified hepatic P-450 (F1). This antiserum recognizes both P-450 (F1) and P-450 (M3). Western immunoblotting using the antiserum demonstrated that female rat brain contains P-450 (F1) but not P-450 (M3), since microsomes from the brain and liver displayed only one immunoreactive band at 50 kD, coinciding with that of P-450 (F1) purified from female rat liver. On the other hand, the male brain has P-450 (M3) but not P-450 (F1), as liver- and brain-derived microsomes produced single band at 49 kD, which represents a mol. wt. identical to that of P-450 (M3) extracted from male rat liver. These results indicate that P-450 (F1)-like immunoreactivity (LI) occurs in the female rat brain, while P-450 (M3)-LI takes place in the male rat brain. Immunocytochemical analysis further demonstrated the detailed cellular localization of these two P-450-LIs in the preoptic area and hypothalamus of female and male rats. Localization of P-450 (F1)-LI in the female rat hypothalamus resembled that of P-450 (M3)-LI in the male rat hypothalamus. Magnocellular neurosecretory neurons in the paraventricular nucleus and supraoptic nucleus were labeled and were found to contain oxytocin but lack vasopressin when serial sections of these areas were analyzed. In addition, groups of immunoreactive cells were seen in the median preoptic nucleus, medial and lateral preoptic area, caudal portion of the bed nucleus of the stria terminalis, lateral hypothalamus at the level of the paraventricular nucleus, periventricular zone from the preoptic area to the paraventricular nucleus, and parvocellular portion of the paraventricular nucleus.     

Demonstration of a sexual dimorphism in the distribution of serotonin-immunoreactive fibers in the medial preoptic nucleus of the rat

The distribution of serotonergic fibers in the medial preoptic nucleus (MPN) and adjacent areas was evaluated with an indirect immunohistochemical method in the normal adult male and female albino rat. Sections through the MPN were processed for immunofluorescence with an anti-serum directed toward serotonin and were counterstained with the fluorescent Nissl stain ethidium bromide. The distribution of serotonin-immunoreactive fibers in the MPN was correlated with cytoarchitectonic features of the nucleus. On the basis of the results, we have subdivided the MPN into three parts: a medial cell-dense part ( MPNm ), a lateral cell-sparse part ( MPNl ), and a central very cell-dense part ( MPNc ) that is embedded in the medial part. The MPNc corresponds to the sexually dimorphic nucleus of the preoptic area identified by Gorski et al. ('80). A marked sexual dimorphism was found in the relative size of each part of the MPN. In the male, the volumes of the cell-dense MPNm and MPNc appear to be notably larger, while in the female more than half of the nucleus is occupied by the cell-sparse lateral part. The MPN as a whole appears to be slightly larger in the male. Each subdivision contains a characteristic pattern of serotonin-immunoreactive fibers. In each sex, the MPN is surrounded by a low to medium density of serotonin-stained fibers, while the MPNl is filled with a dense plexus of varicose immunoreactive fibers. In contrast, the MPNm contains a low density of stained fibers, and the MPNc is virtually devoid of serotonin-stained fibers. Since both the MPNm and the MPNc are larger in the male, a correspondingly larger region of very low serotonin-stained fiber density is found in the male. It appears then that the MPN is a sexually dimorphic complex composed of at least three cytoarchitectonically distinct subdivisions, each of which contains a characteristic density of serotonin-immunoreactive fibers.     

Single unit recording in hypothalamus and preoptic area of estrogen-treated and untreated ovariectomized female rats.

Single unit activity was recorded with micropipettes in the medial hypothalamus and preoptic area of urethane-anesthetized ovariectomized female rats. Some females had received long-term estradiol treatment, while others had been left untreated. In the medial preoptic region and bed nucleus of the stria terminalis, estrogen-treated rats had fewer cells (compared to untreated rats) with recordable spontaneous activity, due primarily to a loss of cells with very slow firing rates. In the basomedial hypothalamus, estrogen-treated rats had more cells (than untreated rats) with recordable spontaneous activity, due primarily to an increase in the number of cells with slow firing rates. Responsiveness of neurons to somatosensory stimulation was generally low. If present it was depressed by estrogen treatment in medial preoptic area and bed nucleus of stria terminalis, while it tended to be elevated by estrogen treatment in medial anterior hypothalamus and basomedial hypothalamus. Differences in the effects of long-term systemic estrogen treatment on medial preoptic neurons compared to basomedial hypothalamus are paralledled by differences in the control of lordosis by these neurons in female rats.     

Testosterone Implants in Specific Neural Sites Activate Female Sexual Behaviour

Sexual behaviour in most female mammals is regulated by oestrogen, often acting synergistically with progesterone. Moreover, the most important neural site of action for oestradiol is the ventromedial nucleus. In the female musk shrew, Suncus murinus, testosterone (T) activates sexual behaviour. Virgin females first engage in copulatory behaviour many hours in advance of follicular development and ovulation, when plasma oestradiol levels are very low. Testosterone, produced by the ovaries and the adrenal glands, must be aromatized centrally to oestradiol to initiate sexual behaviour. To identify the neural sites of action for T, ovariectomized females received unilateral hormone implants containing testosterone propionate. Hormone pellets were placed in 1 of several brain sites including the medial preoptic area and the dorsalmedial hypothalamus (DMH). Implants in either of these 2 sites, but not in the lateral preoptic area, internal capsule, nor anterior hypothalamus stimulated the induction of sexual behaviour. Hormone implants in the ventrolateral hypothalamus resulted in partially receptive animals. Immunocytochemistry was employed to determine which steroid receptors were present in the 2 behaviourally active sites. The medial preoptic area (MPO) and the dorsal and ventromedial hypothalamus both contain many cells that express oestrogen receptor immunoreactivity. A smaller subset of neurons in these regions are immunoreactive for androgen receptors. In summary, testosterone can act specifically in either the MPO or the DMH to induce female sexual behaviour. Both sites contain cells that express oestrogen and androgen receptors. Thus, testosterone may work via one or both of these steroid receptors to regulate behaviour. The results of these studies, and data collected previously, show that female sexual behaviour in this species is regulated by T and that this steroid can induce full sexual behaviour in 2 specific neural sites. Both of the sites of action identified in this experiment are critical for the expression of sexual behaviour in other species. The MPO has been shown to regulate male sexual behaviour via the aromatization of T to oestradiol in rodents and birds. The DMH is just dorsal to the ventromedial nucleus (VMN) which is essential for the regulation of sexual behaviour in female rodents. Further work is needed to determine if the neural circuitry for sexual behaviour in the musk shrew involves either the MPO or the VMN, if both areas are essential, or perhaps, the 2 regions regulate different aspects of female sexual behaviour.     

Stereologic analysis of estrogen receptor alpha (ER alpha) expression in rat hypothalamus and its regulation by aging and estrogen

The estrogen receptor alpha (ERalpha) in the hypothalamus plays important roles in the regulation of reproductive development, physiology, and behavior. However, the expression of the ERalpha may change during aging or in response to varying estrogen levels. The present study measured changes in the numbers of ERalpha-expressing cells in specific hypothalamic and preoptic nuclei of ovariectomized female Sprague-Dawley rats at three ages (young [3-4 months], middle-aged [10-12 months], or old [24-26 months]) and with or without estrogen replacement. Numbers of ERalpha-immunoreactive neurons were quantified in four regions relevant to reproductive function: the anteroventral periventricular nucleus (AVPV), medial preoptic nucleus (MPN), arcuate nucleus (ARH), and ventromedial nucleus (VMN), using an unbiased stereologic approach. In the AVPV and VMN, significant age-related increases in the numbers of ERalpha-expressing cells from the middle-aged to the old group were detected, and no differences were observed in the MPN and ARH, indicating that ERalpha neuron number is maintained or even elevated during aging. No significant effects of estrogen on ERalpha cell number were detected in any of the four regions studied. Therefore, ERalpha cell number in the rat hypothalamus and preoptic area changes with aging in a region-specific manner.     

Projections of the medial preoptic nucleus: a Phaseolus vulgaris leucoagglutinin anterograde tract-tracing study in the rat

The projections of the medial preoptic nucleus (MPN) were examined by making injections of the anterogradely transported lectin Phaseolus vulgaris leucoagglutinin (PHA-L) into the MPN and charting the distribution of labeled fibers. The evidence indicates that the MPN projects extensively to widely distributed regions in both the forebrain and brainstem, most of which also supply inputs to the nucleus. An important neuroendocrine role for the MPN is underscored by its extensive projections to almost all parts of the periventricular zone of the hypothalamus, including the anteroventral periventricular, anterior part of the periventricular, paraventricular (PVH), and arcuate nuclei, and a role in autonomic mechanisms is indicated by projections to such regions as the dorsal and lateral parvicellular parts of the PVH, the lateral parabrachial nucleus, and the nucleus of the solitary tract. Other projections of the MPN suggest participation in the initiation of specific motivated behaviors. For example, inputs to two nuclei of the medial zone of the hypothalamus, the ventromedial and dorsomedial nuclei, may be related to the control of reproductive and ingestive behaviors, respectively, although the possible functional significance of a strong projection to the ventral premammillary nucleus is presently unclear. The execution of these behaviors may involve activation of somatomotor regions via projections to the substantia innominata, zona incerta, ventral tegmental area, and pedunculopontine nucleus. Similarly, inputs to other regions that project directly to the spinal cord, such as the periaqueductal gray, the laterodorsal tegmental nucleus, certain medullary raphe nuclei, and the magnocellular reticular nucleus may also be involved in modulating somatic and/or autonomic reflexes. Finally, the MPN may influence a wide variety of physiological mechanisms and behaviors through its massive projections to areas like the ventral part of the lateral septal nucleus, the bed nucleus of the stria terminalis, the lateral hypothalamic area, the supramammillary nucleus, and the ventral tegmental area, all of which have extensive connections with regions along the medial forebrain bundle. Although the PHA-L method does not allow a clear demonstration of possible differential projections from each subdivision of the MPN, our results suggest that each of them does give rise to a unique pattern of outputs.(ABSTRACT TRUNCATED AT 400 WORDS)     

Efferent Projections from the Ovarian Steroid Receptor-Containing Area of the Ventrolateral Hypothalamus in Female Guinea Pigs

The ventrolateral hypothalamus (VLH) in female guinea pigs includes a subset of neurons which contain estrogen and progestin receptors, and which are implicated in the regulation of female sexual behavior by steroid hormones. However, little is known about where these neurons project, and consequently which other brain areas are involved in sexual behavior in female guinea pigs. The anterograde tracer Phaseolus vulgaris -Leucoagglutinin was used to label efferents from the ovarian steroid receptor-containing part of the VLH. To identify the correct placement of the tracer specifically within the group of neurons containing estrogen receptors, medial hypothalamic sections were also immunostained for estrogen receptors. Forebrain areas receiving dense projections from the ventrolateral hypothalamus included the bed nucleus of the stria terminalis, medial preoptic area, anterior hypothalamic area, anterior ventromedial hypothalamus, and caudal ventrolateral hypothalamus. The midbrain central gray was also heavily labeled. Moderate innervation was observed in the forebrain in the basolateral amygdala, medial preoptic nucleus, lateroanterior hypothalamic nucleus, dorsal hypothalamic areas, posterior hypothalamus, zona incerta, and in the midbrain interspersed among the central and lateral tegmental tracts. The major efferent pathways from the VLH appeared to travel rostrally through the mediobasal hypothalamus and preoptic area, and caudally via the medial thalamic nuclei and periventricular fiber system. These findings are similar to those of previous studies tracing the efferents from the ventromedial nucleus in rats and from the lateral hypothalamus in guinea pigs. Many of these areas that receive input from the steroid receptor rich area within the VLH are likely to be involved in the regulation of female sexual behavior.     

Sex hormone binding globulin stimulates female sexual receptivity

Sex hormone binding globulin (SHBG) is found in the brain and acts directly on plasma membrane-associated receptors in the prostate gland. Infusing SHBG into the medial preoptic area or medial basal hypothalamus of female rats increases their female sexual receptivity. SHBG, SHBG plus estradiol (SHBG-E), and SHBG-E plus oxytocin all significantly increased female sexual receptivity over vehicle or estradiol plus oxytocin infused controls, as measured by lordosis quotients and receptivity scores, at 40, and 90 min after their infusions into the medial preoptic area. When infused into the medial basal hypothalamus, SHBG-E plus oxytocin resulted in significantly increased sexual receptivity 20 and 40 min after infusion when compared to its estradiol plus oxytocin control group. SHBG produced in the brain may be released endogenously to have immediate effects on reproductive physiology and behavior.     

Photoperiodic regulation of vasopressin receptor binding in female Syrian hamsters

During long "summer-like" photoperiods, female Syrian hamsters display a regular 4-day estrous cycle. However, during short "winter-like" photoperiods (<12.5 h of light/day) hamsters become anestrus. Short photoperiod exposure eliminates reproductive behavior but social behaviors such as aggression and scent marking continue to be displayed. In long photoperiods, the types and intensity of social behaviors change as a function of the estrous cycle. For example, aggression and scent marking tend to occur at higher levels on diestrus 1 and diestrus 2 than on proestrus or estrus. Aggression and scent marking may be regulated, at least in part, by changes in the density of arginine vasopressin-V(1a) receptors (V(1a)R). In Experiment 1, it was hypothesized that the density of V(1a)R would change across the estrous cycle in several subcortical regions implicated in the regulation of aggression and scent marking. In Experiment 2, it was hypothesized that exposure to short photoperiod would alter the density of V(1a)R in several regions involved in the regulation of social behavior. Interestingly, there were no dramatic changes in V(1a)R binding across the estrous cycle within any of the neuroanatomical areas measured. However, in hamsters housed in short photoperiod, there were lower levels of V(1a)R binding within the medial preoptic nucleus (MPN), medial preoptic area (MPO), lateral hypothalamus (LH), central nucleus of the amygdala (Ce) and bed nucleus of the stria terminalis (BST) than in hamsters housed in long photoperiod. These data suggest that photoperiodic mechanisms can alter the density of V(1a)R in subset of V(1a) binding sites thought to be involved in the regulation of social behaviors in female hamsters.