Medial Preoptic-anterior Hypothalamic Area Research Articles

Effects of puerarin on estrogen-regulated gene expression in gonadotropin-releasing hormone pulse generator of ovariectomized rats

Effects of puerarin on the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator function is investigated, for the first time, in ovariectomized rats at the level of mRNA expression of estrogen-responsive genes, e.g., estrogen receptor (ER), GnRH and its receptor (GnRHR). Rats were treated orally for 90 days either with a soy-free diet containing two different doses of puerarin (low dose of 600 mg/kg and high dose of 3000 mg/kg) or estradiol benzoate (E2B) at either low dose (4.3 mg/kg) or high dose (17.3 mg/kg). Levels of mRNA expression in the medial preoptic area/anterior hypothalamus (MPOA/AH), mediobasal hypothalamus/median eminence (MBH/ME) and adenohypophysis were measured by quantitative TaqMan® real-time RT-PCR. Plasma levels of luteinizing hormone (LH) and prolactin (PRL) were measured by radioimmunoassay. In the MPOA/AH, both puerarin and E2B decreased ERα mRNA levels without any significant changes in ERβ and GnRH mRNA levels. Both puerarin and E2B did not significantly alter the expression levels of ERα, ERβ and GnRHR in the MBH/ME. E2B exerted significant effects on the down-regulation of adenohypophyseal GnRHR mRNA transcripts and serum LH levels. Puerarin did not cause significant changes in pituitary GnRHR mRNA transcripts and serum LH and PRL levels. This is the first study to demonstrate that in ovariectomized rat models of ovarian hormone deprivation, puerarin acted as a weak estrogen-active compound in the hypothalamic GnRH pulse generator through the downregulation of MPOA/AH ERα mRNA expression.

Lack of anti-androgenic effects of equol on reproductive neuroendocrine function in the adult male rat

Equol (EQ), a metabolite of the soy isoflavone daidzein, has well known estrogenic properties. Data from animal studies suggested that EQ may act also as an anti-androgen. However, data regarding how EQ may affect brain functions like the regulation of neuroendocrine activity and reproductive outcomes in adult male rats are still lacking. We therefore investigated the effects of EQ on sex-steroid regulated gene expression in the brain [medial preoptic area/anterior hypothalamus (MPOA/AH) and medial basal hypothalamus/median eminence (MBH/ME)], pituitary, and prostate as a reference androgen-dependent organ. Furthermore reproductive outcomes were evaluated. The anti-androgen flutamide (FLUT) served as reference compound. Male rats (n=12 per group) were treated by gavage for 5days with either EQ (100 or 250mg/kgBW/day), or FLUT 100mg/kgBW/day. All vehicle- and EQ-treated males showed successful reproductive outcomes, whereas FLUT-exposed males had severe reproductive impairments resulted in infertility. FLUT decreased relative weights of prostate, seminal vesicles and epididymides, and increased serum levels of luteinizing hormone, follicle-stimulating hormone, testosterone and 5α-dihydrotestosterone without altering prolactin levels, whereas EQ exerted opposite effects. Both EQ and FLUT decreased gonadotropin releasing hormone (GnRH) expression in the MPOA/AH. Only FLUT upregulated levels of GnRH receptor expression both in the MBH/ME and pituitary. While EQ downregulated the hypothalamic ERα and ERβ expressions, but FLUT did not. In the prostate, only FLUT upregulated both ERα and AR mRNA expression levels. Taken together, our findings are the first data that EQ did not induce anti-androgenic effects on brain, prostate and male reproductive parameters, however, estrogenic neuroendocrine and reproductive effects of EQ were observed.

Seasonal regulation of social communication by photoperiod and testosterone: Effects of arginine–vasopressin, serotonin and galanin in the medial preoptic area–anterior hypothalamus

Arginine–vasopressin (AVP) activation of V1a AVP receptors in the medial preoptic area–anterior hypothalamus (MPOA–AH) plays a critical role in the control of a form of social communication called flank marking. The ability of AVP and V1a receptors in the MPOA–AH to regulate flank marking is modulated by a number of factors including galanin (GAL), serotonin (5-HT) and gonadal hormones. More recent studies suggest that there may be seasonal differences in the number or type of AVP receptors in the MPOA–AH controlling flank marking or in the modulation of these mechanisms by GAL or 5-HT. The purpose of the present study was to examine how exposure to “summer-like” and “winter-like” photoperiods alters the neural mechanisms regulating flank marking. These studies confirmed that V1a AVP receptors, but not V1b AVP receptors play a significant role in controlling flank marking in the MPOA–AH in both LP and SP-housed hamsters. These studies also found that there were no significant differences between hamsters housed in LP and SP in the ability of serotonin 5-HT1a/7 and 5-HT1b receptor agonists injected into the MPOA–AH to inhibit odor-induced flank marking. Finally, these studies found that GAL could significantly inhibit the ability of AVP to stimulate flank marking in SP-housed hamsters and that there were no differences between hamsters housed in LP and SP in the ability of a GAL antagonist to inhibit AVP-induced flank marking. The present study confirms that V1a receptors are critical for AVP regulation of flank marking and that photoperiodic mechanisms do not alter the modulation of flank marking by GAL or 5-HT in the MPOA–AH.

Inactivation of the medial preoptic area/anterior hypothalamus by lidocaine reduces male sexual behavior and sexual incentive motivation in male rats

Permanent bilateral lesions of the medial preoptic area anterior hypothalamus (MPOA/AH) produce a drastic inhibition of male sexual behavior in all species studied to date. The present experiment was designed to evaluate if temporal inactivation of the MPOA/AH by infusions of lidocaine also inhibits sexual behavior in male rats. This would allow us to rule out the possibility that the behavioral effects observed after damage of the MPOA/AH could be associated with plastic changes induced by the lesion in other brain regions. We also evaluated sexual incentive motivation in males after the infusion of lidocaine in a test in which copulation is not possible but where males maintain approach behavior to the estrous females despite repeated testing. The percentage of animals displaying mounts, intromissions and ejaculation was significantly reduced while mount and intromission latency were prolonged after infusion of lidocaine. No changes were observed in sexual behavior after infusion of lidocaine in animals with cannulae outside the MPOA/AH suggesting that the inhibitory effects are specific to this brain region. Sexual incentive motivation was also affected by administration of lidocaine. Males consistently showed a clear preference for the sexually receptive female except when infused with lidocaine. After the infusion of the compound a significant reduction in the time spent in the incentive zone of the stimulus female was observed. These results support the hypothesis that neurons of the MPOA/AH are involved in the control of male sexual motivation.

Central tegmental field and sexual behavior in the male rat: Effects of neurotoxic lesions

The medial preoptic area/anterior hypothalamus (MPOA/AH) is a key structure in the control of male sexual behavior. This area has reciprocal connections with mesencephalic and brainstem structures including the central tegmental field (CTF). It has been suggested that the CTF receives somatosensory information generated in the genitals promoting activation of the MPOA/AH. In the present study we evaluated the effects of bilateral neurotoxic lesions of the CTF upon male rat sexual behavior. We also explored the effects of these lesions on sociosexual behaviors, partner preference, sexual incentive motivation and motor execution. Tests were performed before and after bilateral quinolinic acid infusions. The lesion was evaluated by quantifying neuronal nuclei (Neu-N) and by the presence of glial fibrillary acidic protein (GFAP) immunohistochemistries. A significant reduction in the percentage of animals displaying mounts, intromissions, and ejaculations was observed in the bilateral and misplaced lesion groups 1 week after the lesion. In the second week post-lesion, only animals with bilateral damage of the CTF showed a significant reduction in sexual behavior. In the third post-lesion test, the percentage of animals displaying sexual behavior returned to control levels. The frequency of pursuit and self-grooming was reduced, and genital exploration was increased after the lesion. Partner preference and sexual incentive motivation were not affected by the lesion suggesting that the CTF is not involved in the appetitive aspects of sexual behavior. Mount, intromission, and ejaculation latency were increased in animals with damage of the CTF and in animals with lesions outside this region. Motor execution was also affected in both groups, suggesting that alterations in latencies could be associated with damage not specific to the CTF.

The role of the dorsolateral tegmentum in the control of male sexual behavior: A reevaluation

The medial preoptic area/anterior hypothalamus (MPOA/AH) plays a key role in the control of male sexual behavior. Independently of the type, MPOA/AH lesions permanently eliminate male sexual behavior in the rat. The MPOA/AH projects among other structures to the dorsolateral tegmentum (DLT). Bilateral electrolytic lesions of the DLT or the unilateral electrolytic destruction of the MPOA/HA combined with a contralateral electrolytic lesion of the DLT eliminate male sexual behavior. In the present experiment, we evaluated if neurotoxic lesions of the DLT produce the same behavioral deficits as those observed after electrolytic lesions. This would allow us to evaluate if neurons of the DLT or the fibers passing through this area are important in the control of male sexual behavior. To this aim, sexually experience male rats were tested for socio-sexual behavior, partner preference and motor execution in order to determine if the possible behavioral changes could be attributed to alterations in sexual motivation or motor execution. One week after the bilateral DLT lesions the animals were evaluated in the same behavioral tests. The lesions were identified by glial fibrillary acidic protein (GFAP) and neuronal nuclear protein (Neu-N) immunohistochemistry. No significant consistent effects upon sexual behavior were observed in any of the groups, including the group with clear bilateral damage of the DLT. A reduction in the percentage of males displaying ejaculation in the first post-lesion test was observed for all groups injected with quinolinic acid. No effects upon partner preference or motor coordination were observed after the lesion in any of the groups. The lack of effect of DLT neurotoxic lesions upon mating suggests that neurons of this structure are not involved in the control of male sexual behavior.

Estradiol Control of Expression and Levels of Estradiol-Binding Proteins in the Medial Preoptic Area, Medial Hypothalamus and Pituitary

The brains of mammals have at least three estradiol-binding proteins: estradiol receptor-α (ERα), ERβ, and sex hormone-binding globulin (SHBG). In this study we compare the effects of estradiol treatment on the expression of mRNA for these three estradiol-binding proteins in two reproductively important brain areas, the medial preoptic area-anterior hypothalamus (MPOA-AH) and medial hypothalamus (MH) as well as in the hippocampus in ovariectomized rats, using the reverse transcriptase-polymerase chain reaction (RT-PCR). We also used surface-enhanced laser desorption ionization time of flight (SELDI-TOF) mass spectrometry (MS) to analyze the effects of estradiol in ovariectomized rats on SHBG levels in the MPOA-MH as well as the neurohypophysis. In vivo estradiol treatment in ovariectomized rats eliminated or significantly reduced expression of all three estradiol-binding proteins in both the MPOA-AH and MH. This change in ERα, ERβ, and SHBG expression did not occur in the hippocampus. Both Northern blot and DNA sequence analysis confirmed the results of the RT-PCR for SHBG. SELDI-TOF MS analysis demonstrated that in vivo estradiol treatments resulted in dramatically decreased levels of SHBG in the hypothalamus and that a reduction in SHBG mRNA by estradiol treatment also resulted in a reduction in SHBG protein levels. Estradiol treatment also eliminated detectable SHBG from the neurohypophysis, suggesting that estradiol controls SHBG levels in this release site. That in vivo estradiol treatments had the same inhibitory effects on mRNA levels for SHBG and both ERs suggests similar translational control mechanisms for all three steroid-binding proteins in the brain. That estradiol treatments also reduced pituitary SHBG suggests that such treatment releases SHBG from the neurohypophysis.

Medial preoptic area/anterior hypothalamus and sexual motivation.

The medial preoptic area/anterior hypothalamus (MPOA/AH) is a brain site derived from proliferative zones from the diencephalon and telencephalon. It is probably this characteristic that makes this brain region participate in different physiological and behavioral functions. The present review addresses the role of the MPOA/AH in the control of male sexual behavior. It is clear that the MPOA/AH is a crucial site in the control of sexual behavior in males of all species studied to date. But although many different publications have followed the contribution of Heimer and Larsson there is no agreement as to what is specifically the role of the MPOA/AH in sexual behavior. At least three hypotheses have been presented. The first one suggests that this brain region is involved in the consummatory aspects (execution) of sexual behavior. The second indicates that the MPOA/AH is involved in the appetitive components (motivation) of masculine sexual behavior. The third hypothesis considers that MPOA/AH neurons are involved in the regulation of consummatory and appetitive aspects of sexual behavior. From the literature reviewed, it will become evident that the evidence supporting a role of the MPOA/AH in the execution of sexual behavior is based on a number of limited studies not easy to interpret. On the other hand, several lines of evidence using a variety of methodologies support the notion that the MPOA/AH is involved in the motivational aspects of male sexual behavior.

Sex hormone binding globulin facilitates female sexual receptivity except when coupled to dihydrotestosterone

Sex hormone binding globulin (SHBG) is produced in brain where it is often co-localized with oxytocin. Infusions of SHBG into the medial preoptic area–anterior hypothalamus facilitate female sexual receptivity. SHBG has receptors on plasma membranes of the prostate gland where binding of the 5α-reduced androgen dihydrotestosterone (DHT) by SHBG acts as an antagonist on SHBG receptors. This study attempted to determine whether pre-coupling DHT to SHBG would inhibit SHBG-induced facilitation of female sexual receptivity. Ovariectomized rats were injected daily with 0.75 μg estradiol benzoate for 3 days. On the fourth day after a pre-infusion baseline behavioral test animals were infused with 1 μl per side through bilateral cannulae with SHBG (1.77×10 −6 M), SHBG coupled to DHT (SHBG–DHT; 1.66×10 −6 M DHT), with DHT alone or with artificial cerebrospinal fluid vehicle. As before, SHBG significantly increased female sexual receptivity when infused into the medial preoptic area–anterior hypothalamus. Rats infused with SHBG–DHT had significantly lower sexual receptivity. Therefore, whereas SHBG in the medial preoptic area facilitated female sexual behavior, SHBG coupled to DHT did not. DHT itself did not significantly affect sexual receptivity. Pre-coupling DHT to SHBG eliminated the facilitative effect of SHBG on female sexual receptivity just as DHT inhibits SHBG activity at prostate SHBG receptors suggesting that central receptors for SHBG are similar to those demonstrated in the periphery.

Vasopressin into the preoptic area increases grooming behavior in mice

In mice, the neuropeptide arginine-8-vasopressin (AVP) induces excessive grooming, scratching, and hyperactivity when administered intracerebroventricularly. In hamsters, AVP infusion into the medial preoptic area/anterior hypothalamus (MPOA/AH) increases flank marking and flank mark grooming. We measured the behavioral effects of administration of AVP (0, 1, and 10 ng/250 nl) into the preoptic area (POA) of male C57BL/6 mice. Administration of AVP into the POA induced robust effects on grooming, including increased hindleg scratching and face washing. Rearing and olfactory investigation were inhibited by AVP into the POA. These findings indicate that the POA is one site in which AVP induces grooming behavior in mice.

Evidence of sex hormone binding globulin binding sites in the medial preoptic area and hypothalamus.

We have demonstrated a high density of both radiolabeled progesterone and estradiol conjugated to bovine serum albumin binding sites in the medial preoptic area and hypothalamus. Infusions of sex hormone binding globulin into the medial preoptic area of rats increased their female sexual receptivity similarly to the effect of estradiol conjugated to bovine serum albumin, suggesting sex hormone binding globulin acts at binding sites for estradiol conjugated to bovine serum albumin. In this study sex hormone binding globulin was used to displace radiolabeled progesterone conjugated to bovine serum albumin from plasma membrane fractions from the medial preoptic area-anterior hypothalamus and medial basal hypothalamus of ovariectomized rats injected with either 5 microg estradiol benzoate or sesame oil vehicle. We found that sex hormone binding displaced radiolabeled progesterone conjugated to bovine serum albumin in both areas and that in vivo estradiol treatment greatly increased the relative displacement by sex hormone binding globulin in the medial preoptic area-anterior hypothalamus. We interpret these data as indicating the presence of sex hormone binding globulin receptors in brain plasma membranes and further suggest that endogenous steroid conditions may alter these receptors.

Radioligand assays for oestradiol and progesterone conjugated to protein reveal evidence for a common membrane binding site in the medial preoptic area-anterior hypothalamus and differential modulation by cholera toxin and GTPgammaS.

In this study membrane oestradiol (E) binding sites in the medial preoptic area-anterior hypothalamus (MPOA-AH) of ovariectomized (OVX) rats were characterized using standard radioligand binding techniques employing E conjugated to bovine serum albumin (BSA) at position 6 and radiolabeled with 125I (E-6-[125I-BSA]). In previous studies binding of a radioactive conjugate of progesterone (P) and BSA (P-3-[125I-BSA]) was examined using the same membrane preparation. E-6-[125I-BSA] binding was linear across a tissue concentration range of 0.005-0.02 mg protein/0.1 ml of membrane suspension. An association T1/2 of 9.5 min and a dissociation T1/2 of 52.1 min for E-6-[125I-BSA] were derived from kinetic experiments. Competition binding experiments revealed high (Ki=0.63+/-(0.50 nM) and low (Ki=161.5(96.5 nM) affinity binding sites for E-6-[125I-BSA], demonstrating different binding parameters than shown in our previous work for P-3-[125I-BSA] binding. Further studies on MPOA-AH membranes treated with cholera toxin (CTX) and GTPgammaS suggested that E-6-BSA binding sites are associated with G proteins. E-6-[125I-BSA] binding demonstrated both high-and low-affinity sites. GTPgammaS added to the assay reduced both E-6-[125I-BSA] and P-3-[125I-BSA] binding suggesting that G proteins are associated with both binding sites. Extensive analysis of both E-6-[125I-BSA] and P-3-[125I-BSA] binding sites demonstrated a reciprocal relationship such that high-affinity E-6-[125I-BSA] binding sites exhibit low affinity for P-3-[125I-BSA] and low-affinity E-6-[125I-BSA] binding sites exhibit high affinity for P-3-[125I-BSA]. Preincubating membranes with CTX or GTPgammaS reduced high-affinity E-6-[125I-BSA] binding and enhanced high-affinity P-3-[125I-BSA] binding. These results suggest that, in the MPOA-AH, membrane steroid binding sites exist in two interconvertible conformations that preferentially bind either E-6-BSA or P-3-BSA, depending on their association with a G protein. Additional studies with free steroids revealed that: (1) oestrogens (17beta-oestradiol, diethylstilbestrol) as well as synthetic oestrogen antagonists tamoxifen and ICI 182 780 displaced P-3-[125I-BSA] further suggesting a relationship between membrane binding sites for E and P-3-[125I-BSA] binding sites; and (2) treatment of OVX rats with E decreased displacement by P-3-BSA and increased displacement by ICI 182,780 and tamoxifen suggesting these antagonists affect membrane P-3-[125I-BSA] binding sites after in-vivo E treatment. The membrane binding sites for E and P demonstrate interrelationships not demonstrated by their nuclear receptors.

Androgen receptor messenger ribonucleic acid in brains and pituitaries of male rhesus monkeys: studies on distribution, hormonal control, and relationship to luteinizing hormone secretion.

Because the distribution and hormonal regulation of the androgen receptor (AR) mRNA in brains and pituitaries of adult rhesus monkeys have not been studied, we cloned and sequenced a 329-base pair segment of the 5' coding region of the rhesus AR cDNA. Monkey AR cDNA was 99% identical with the human sequence and 96% homologous with the rat sequence. Using a ribonuclease protection assay, we studied the distribution and regulation of AR mRNA in brains and anterior pituitary glands of three groups of male rhesus monkeys: intact (n = 3), castrated (Cx, n = 4), and Cx treated with testosterone (n = 6). Serum testosterone levels of Cx males treated with testosterone differed significantly (p < 0.05) in the morning but not in the evening hours from those in intact controls. Serum LH concentrations were significantly suppressed (p < 0.05) in both morning and evening serum samples of testosterone-treated males compared to intact controls. We found the highest concentrations of AR mRNA in the medial basal hypothalamus, the bed nucleus of the stria terminalis, the medial preoptic area-anterior hypothalamus, and the lateral dorsomedial hypothalamus. Intermediate amounts were found in the septum and amygdala. Low amounts were found in the hippocampus, cingulate cortex, parietal cortex, and cerebellum. The anterior pituitary gland also contained a large amount of AR mRNA. Surprisingly, neither Cx for 3 wk nor Cx plus testosterone replacement for 3 wk significantly affected AR mRNA in any brain area or in the pituitary gland. The present study demonstrates that the effectiveness of testosterone as a regulator of LH secretion in male monkeys is not related to changes of AR mRNA in the brain or pituitary gland. It appears that AR mRNA in the monkey brain and pituitary gland is not regulated at the transcriptional level by androgen.

Conjugated Estradiol Increases Female Sexual Receptivity in Response to Oxytocin Infused into the Medial Preoptic Area and Medial Basal Hypothalamus

The ovarian steroid estradiol (E) has been found to increase both receptor affinity and release of the neuropeptide oxytocin (OT) in plasma membrane preparations. Therefore, we hypothesized that E conjugated to bovine serum albumin at position 6 (E-6-BSA) would increase behavioral responsiveness to OT. Preliminary results showed that 200 ng/μl of E-6-BSA increased sexual receptivity slightly, but not significantly. Therefore, this dose was used as a subthreshold dose to test whether it would increase sexual responsiveness when infused in combination with 100 ng/μl OT. After recovery from cannula implantation surgery animals were injected with 0.5 μg E benzoate daily for 3 days before testing. On the fourth day, after a baseline preinfusion test rats were infused bilaterally with E-6-BSA alone or with OT, OT with BSA, or conjugated progesterone, luteinizing hormone-releasing hormone equimolar to OT alone, or with E-6-BSA or conjugated progesterone alone. When infused into either the medial preoptic area-anterior hypothalamus or the medial basal hypothalamus the combination of OT and E-6-BSA significantly increased sexual receptivity over receptivity after artificial cerebrospinal fluid control infusions. Neither bilateral infusions of OT in combination with conjugated progesterone nor E-6-BSA in combination with luteinizing hormone-releasing hormone enhanced sexual receptivity. Results presented here strongly support the conclusion that some of the effects that E has in sensitizing brain systems to the facilitating effects of OT occur at the membrane level in the medial preoptic area-anterior hypothalamus and medial basal hypothalamus.

Injection of Oxytocin into the Medial Preoptic–Anterior Hypothalamus Increases Ultrasound Production by Female Hamsters

Recent studies of hamsters have documented the facilitation of lordosis and other sociosexual responses by injections of oxytocin (OXT) into the medial preoptic area-anterior hypothalamus (MPOA-AH). These data suggest the regulation of social interaction and bonds by OXT. In turn, this suggests that OXT could act in the MPOA-AH to control other behaviors involved in the initiation or maintenance of social contact, including the ultrasonic vocalizations that female hamsters use to alert and attract potential mates. To test this possibility, we compared the ultrasound rates of 11 naturally estrous hamsters before and after injections of OXT (200 ng/200 nl of saline) or saline (200 nl) into the MPOA-AH. The data revealed a clear facilitation of ultrasound rate 30 min after OXT treatment. This result suggests the modulation of ultrasound rate by endogenous OXT acting within the MPOA-AH. It extends the range of social behaviors sensitive to control by OXT and supports the possibility that OXT acts within the MPOA-AH to facilitate a variety of behaviors involved in the establishment or maintenance of the social interactions required for successful reproduction. At the same time, these data extend earlier observations linking ultrasound production to the MPOA-AH, and begin to describe the peptidergic mechanisms controlling this form of reproductive behavior.

Site-Specific Opioid Receptor Blockade Allows Prepubertal Guinea Pigs to Display Progesterone-Facilitated Lordosis

Ovariectomized (OVX) juvenile guinea pigs (∼3 weeks old) rarely display steroid-induced sexual receptivity. Systemic administration of the opioid receptor antagonist naloxone enhances the display of progesterone-facilitated lordosis in prepubertal females, suggesting that endogenous opioids tonically inhibit the expression of sexual receptivity at this age. This study was designed to ascertain the neural site(s) at which naloxone injection would stimulate lordosis in juvenile guinea pigs. Hartley guinea pigs were OVX at 10–11 days of age and 2–6 days later implanted with bilateral cannulae aimed at the medial preoptic area/anterior hypothalamus (MPOA/AH), ventrolateral hypothalamus/ventromedial hypothalamus (VLH/VMH), or mesencephalic central gray (MCG). At 21–23 days of age, following administration of estradiol benzoate (10 μg) and progesterone (0.5 mg), naloxone (100 ng/side) or 0.9% saline was injected through the cannulae and the guinea pigs were tested for the display of lordosis. The MPOA/AH was the only site at which application of naloxone reliably elicited lordosis (87% positive response vs 12% for saline). Few females (<17%) displayed lordosis following injections of naloxone or saline into the VLH/VMH or MCG. A second experiment demonstrated that the stimulation of lordosis following MPOA/AH naloxone application was prevented by prior injection of the opioid agonist morphine (500 ng/side) at the same site. These data support the hypothesis that endogenous opioids acting in the MPOA/AH, but not the VLH/VMH or MCG, tonically inhibit the display of progesterone-facilitated lordosis in prepubertal guinea pigs.

Further studies on the suppression of luteinizing hormone release due to activation of medial preoptic-anterior hypothalamic area mu-opioid receptors.

Our laboratory has previously demonstrated that specific activation of mu-opioid receptors in the medial preoptic-anterior hypothalamic area with [D-Ala2, N Me-Phe4, Gly-ol5]-enkephalin (DAGO) suppresses luteinizing hormone secretion in the ovariectomized rat [29]. In the present study, three experiments were undertaken to ascertain whether changes in the activity of norepinephrine or dopamine neurons modulate the decrease in luteinizing hormone release in response to DAGO. The first experiment utilized push-pull perfusion in conjunction with HPLC to assess in vivo norepinephrine release in the medial preoptic-anterior hypothalamic area in response to perfusion of this site with DAGO (5 micrograms/h). DAGO significantly decreased luteinizing hormone release, but perfusate norepinephrine levels did not change. In the second experiment, push-pull perfusion in the medial preoptic-anterior hypothalamic area with cerebrospinal fluid (CSF) or CSF containing DAGO was done in rats pretreated with the norepinephrine synthesis inhibitor, FLA-63. This drug pretreatment had no effect on the DAGO-induced suppression of luteinizing hormone secretion. In experiment 3, push-pull perfusion in the medial preoptic-anterior hypothalamic area with CSF followed by CSF containing DAGO was done in rats pretreated with vehicle, or a dopamine receptor antagonist, either pimozide or d-butaclamol. Neither dopamine receptor antagonist had any effect on the DAGO-induced suppression of luteinizing hormone release. Thus, these studies do not support a role for alterations in the activity of norepinephrine or dopamine neurons in mediating the suppression of luteinizing hormone release in response to activation of mu-opioid receptors in the medial preoptic-anterior hypothalamic area in the ovariectomized rat.

Characterization of progesterone-3-[ 125I-BSA] binding sites in the medial preoptic area and anterior hypothalamus

In this study we utilized radiolabeled progesterone (P) conjugated to bovine serum albumin (BSA) at position 3 (P-3-[ 125I-BSA]) to examine steroid receptors in membrane fractions from the medial preoptic area-anterior hypothalamus (MPOA-AH) of ovariectomized (OVXed) rats. In the MPOA-AH binding of P-3-[ 125I-BSA] was linear across a tissue concentration range of 0.005 to 0.02 mg protein/0.1 ml of membrane suspension. Kinetic experiments revealed an association t 1/2 of 51.4 min and a dissociation t 1/2 of 122.5 min for P-3-[ 125I-BSA] at 0°C. Analysis of data from competition binding experiments using P-3-BSA revealed high- and low-affinity binding sites in the MPOA-AH. Involvement of MPOA-AH binding sites with a G-protein was suggested by a reduction of P-3-[ 125I-BSA] binding in the presence of the non-hydrolyzable GTP analog GTPγS but not ATPγS. In addition, if homogenates from the MPOA-AH were preincubated with 10 −5 M of the G-protein antagonist cholera toxin for 30 min at 37°C, competition binding data indicated only high-affinity binding sites. Once daily injections of OVXed rats with 4 mg P for 12 days significantly increased the density of P-3-[ 125I-BSA] binding sites in the MPOA-AH. This treatment did not affect P-3-[ 125I-BSA] binding in the dorsal tectum, medial basal hypothalamus, ventral tegmental area or the thymus.

Effects of testosterone on the behavioral response to arginine vasopressin microinjected into the central gray and septum

Arginine vasopressin (AVP) plays an important role in the control of a gonadal hormone-dependent communicative behavior in the Syrian hamster ( Mesocricetus auratus) called flank marking. Previous studies have shown that gonadal hormones alter the amount of flank marking stimulated by the microinjection of AVP into the medial preoptic area-anterior hypothalamus (MPOA-AH). The purpose of the present study was to determine if testicular hormones alter the amount of flank marking stimulated by the microinjection of AVP into two other sites involved in the control of flank marking, the lateral septum-bed nucleus of the stria terminalis (LS-BNST) and the central gray. The data of the present study indicate that testicular hormones may influence the amount of AVP-stimulated marking in the central gray and LS-BNST; however, these effects are subtle and appear to occur primarily at high concentrations of AVP. When taken together with previous studies, these data indicate that gonadal hormones have greater effects on AVP-stimulated marking in the MPOA-AH than in the LS-BNST or central gray.

Effects of hypothalamic lesions on courtship and copulatory behavior in sexual and unisexual whiptail lizards

Lesions of the anterior hypothalamus-medial preoptic area (AH-MPOA) impair courtship and copulatory behavior in male Cnemidophorus inornatus, a sexually reproducing species of whiptail lizard, and male-like pseudosexual behaviors in C. uniparens, an all-female species. These results suggest that, since C. inornatus is a direct ancestor to C. uniparens, the neural structures responsible for mediating male-typical courtship and copulatory behavior in male whiptails are conserved in the evolution of the all-female parthenogen.