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Sexual stimulation induces Fos immunoreactivity within GnRH neurons of the female rat preoptic area: interaction with steroid hormones.

We have shown previously that sexual stimulation (copulation with intromission or vaginocervical stimulation) induces c-fos mRNA and Fos-like immunoreactivity (IR) within estrogen-concentrating and nonconcentrating regions of the female rat forebrain, including regions that contain gonadotropin-releasing hormone (GnRH) neurons in septum and anterior preoptic area. The overall induction of Fos-like IR within these regions was specific to afferent sensory stimulation and did not require treatment with estrogen and progesterone. Because vaginocervical stimulation facilitates lordosis and increases the release of luteinizing hormone, the present study examined whether hormone treatment that promotes sexual receptivity, with or without sexual stimulation, increases Fos-like IR specifically within GnRH-containing neurons. Sexually experienced ovariectomized rats were administered estradiol benzoate (10 micrograms) 48 h and progesterone (500 micrograms) 4 h before either 1 h of paced copulation with a sexually vigorous male, 50 vaginocervical stimulations with a glass rod distributed over 1 h, or no stimulation. Control rats received injections of the oil vehicle. Fos-like IR was found within a significant number of GnRH-positive neurons in the anterior preoptic area caudal to the organum vasculosum following copulation with intromission or vaginocervical stimulation as compared with no stimulation. Although few GnRH cells coexpressed Fos following hormone treatment alone, this treatment enhanced the number of GnRH neurons that coexpressed Fos following vaginocervical stimulation as compared with the effect of vaginocervical stimulation in oil-treated rats. Together, these data indicate that estrogen and progesterone can augment the responsiveness of certain GnRH neurons to vaginocervical stimulation, consistent with the effects of sexual activity on GnRH release.

Animals↗

Ontogeny of sex differences in the mammalian hypothalamus and preoptic area.

1. There are numerous sites in the nervous system where steroid hormones dramatically influence development. Increasing interest in mechanisms in neural development is providing avenues for understanding how gonadal steroids alter the ontogeny of these regions during sexual differentiation. 2. An increasing number of researchers are examining effects of gonadal steroids on neurite outgrowth, cell differentiation, cell death, cell migration, and synaptogenesis. The interrelated timing of these events may be a key aspect influenced by gonadal steroids throughout development. 3. The preoptic area and hypothalamus are characteristically heterogeneous in terms of cell type (e.g., different neuropeptides) and cell derivation. Perhaps a major reason for the ontogeny of sexual differences in the preoptic area and hypothalamus lies in the convergence of many different cell types from diverse sources (i.e., proliferative zones surrounding the lateral and third ventricles, and the olfactory placodes) that can be influenced in an interactive manner by gonadal steroid mechanisms. 4. The characterization of multiple mechanisms (e.g., trophic, migratory, apoptotic, fate, etc.,) that contribute to permanent changes in brain structure and ultimately function is essential for unraveling the process of sexual differentiation.

Animals↗

Microinjection of galanin-like peptide into the medial preoptic area stimulates food intake in adult male rats.

Galanin-like peptide (GALP) is a neuropeptide implicated in the regulation of feeding behaviour, metabolism and reproduction. GALP is an endogenous ligand of the galanin receptors, which are widely expressed in the hypothalamus. GALP is predominantly expressed in arcuate nucleus (ARC) neurones, which project to the paraventricular nucleus (PVN) and medial preoptic area (mPOA). Intracerebroventricular or intraparaventricular (iPVN) injection of GALP acutely increases food intake in rats. The effect of GALP injection into the mPOA on feeding behaviour has not previously been studied. In the present study, intra-mPOA (imPOA) injection of GALP potently increased 0-1-h food intake in rats. The dose-response effect of imPOA GALP administration on food intake was similar to that previously observed following iPVN administration. The effects of GALP (1 nmol) or galanin (1 nmol) on food intake were then compared following injection into the PVN, mPOA, ARC, dorsal medial nucleus (DMN), lateral hypothalamus and rostral preoptic area (rPOA). GALP (1 nmol) increased food intake to a similar degree when injected into the imPOA or iPVN, but produced no significant effect when injected into the ARC, DMN, lateral hypothalamus or rPOA. Similarly, galanin (1 nmol) significantly increased food intake following injection imPOA and iPVN. However, the effect was significantly smaller than that following administration of GALP (1 nmol). Galanin also had no significant effect on food intake when administered into the ARC, DMN, lateral hypothalamus and rPOA. These data suggest that the mPOA and the PVN may have specific roles in mediating the orexigenic effect of GALP and galanin.

Animals↗

Effects of chemical lesion of the ventral noradrenergic bundle or the medial preoptic area on preovulatory LH release in rats.

The noradrenergic innervation of the medial preoptic area (MPO) and the hypothalamus is provided by mesencephalic neurons via the ventral noradrenergic tract. Fibers of these neurons emerge through the MPO. Bilateral microinjections of 6-OHDA into the ventral noradrenergic bundle (VNB) depletes large parts of the diencephalon of norepinephrine (NE) and dopamine (DA). Since the total hypothalamic DA content is of intrahypothalamic origin, 6-OHDA injection into the VNB does not reduce hypothalamic DA content. Similarly microinjections of 6-OHDA into the MPO reduces hypothalamic and preoptic NE content without altering NE concentrations in other diencephalic structures. Microinjections of 6-OHDA and of the carrier solution of 6-OHDA into the VNB or into the MPO of female rats with regular estrous cycles results in a slight disturbance of the cyclic activity for few days. Within 1--4 days normal cyclic activity is resumed. Preovulatory LH release is substantially reduced 8--12 days after injection of 6-OHDA into the VNB or into the MPO. On the basis of these and previous results it is concluded that the availability of NE in the MPO is an important factor in determining the hight of the preovulatory LH surge.

Animals↗

Distribution of preprodynorphin mRNA and dynorphin-a immunoreactivity in the sheep preoptic area and hypothalamus.

Endogenous opioid peptides (EOP) are important modulators in a variety of neuroendocrine systems, including those mediating reproduction, energy balance, lactation, and stress. Recent work in the ewe has implicated the EOP, dynorphin (DYN), in the inhibitory effects of progesterone on pulsatile gonadotropin releasing hormone secretion. Although DYN is involved in a number of hypothalamic functions in the sheep, little is known regarding the localization of preprodynorphin (PPD) expression and its major product DYN A (1-17). In this study, we determined the distribution of PPD mRNA and DYN A-containing cell bodies in the brains of ovary-intact, luteal ewes. To detect PPD mRNA, an ovine PPD mRNA was subcloned by reverse transcription-polymerase chain reaction from sheep hypothalamus and used to create a (35)S-labeled riboprobe for in situ hybridization. Neurons that expressed PPD mRNA and DYN A immunoreactivity were widely distributed in the ovine preoptic area and hypothalamus. PPD mRNA-expressing cells were seen in the supraoptic nucleus, paraventricular nucleus, preoptic area, anterior hypothalamus area, bed nucleus of the stria terminalis, ventromedial nucleus (VMN), dorsomedial nucleus of the hypothalamus, and the arcuate nucleus. All of these regions also contained DYN A-positive cell bodies except for the VMN, raising the possibility that PPD is preferentially processed into other peptide products in the VMN. In summary, based on the expression of both mRNA and peptide, DYN cells are located in a number of key hypothalamic regions involved in the neuroendocrine control of homeostasis in sheep.

Animals↗

Effects of preoptic area lesions on muramyl dipeptide-induced sleep and fever.

Lesions of the preoptic area of rabbits caused hyperthermia, although they did not alter sleep-coupled changes in brain temperature, and reduced slow-wave sleep, rapid eye movement sleep, and amplitudes of electroencephalographic slow waves during slow wave sleep. The direction and magnitude of the muramyl dipeptide-induced responses were similar before and after the lesions, although the baseline values upon which the changes were superimposed had shifted.

Acetylmuramyl-Alanyl-Isoglutamine↗

Estrogen increases GAP-43 (neuromodulin) mRNA in the preoptic area of aged rats.

Estrogen has been shown to affect the growth, differentiation, and survival of brain neurons and to modulate processes involved in synapse formation and connectivity. These trophic effects are diminished with aging as secretion of estrogen declines. The growth associated protein GAP-43 is found concentrated in axonal growth cones and is implicated in neuronal growth and regeneration. Previous studies have established that expression of GAP-43 can be modulated by estrogen in the preoptic area of developing and adult rat brain. This study was undertaken to determine whether this estrogenic regulation of GAP-43 mRNA is retained in aged rat brain. Young (3 months) and aged (24 months) rats were ovariectomized to remove endogenous estrogen and GAP-43 mRNA in the preoptic area was evaluated using in situ hybridization to compare estrogen and vehicle treatments between age groups. The results demonstrate an age-related decline in GAP-43 mRNA hybridization signal that can be restored to levels comparable to that seen in young animals with estrogen treatment.

Age Factors↗

Effects of ethanol injection to the preoptic area on sleep and temperature in rats.

Bilateral microinjection of ethanol to the preoptic area of rats causes a dose-dependent hypnotic effect at doses that do not affect brain temperature. Rats were polygraphically recorded for 6 h, and brain temperature was recorded every 10 min for 3 h. Administration of 0.047 mumol ethanol increased total sleep during the 3-6 h interval, while 0.24 and 0.47 mumol ethanol increased deep slow-wave sleep (SWS2) and total sleep during the 3-6- and 0-6-h intervals. The 0.24-mumol ethanol microinjections also increased REM sleep for the 0-6-h interval. No changes in sleep latencies were observed.

Animals↗

Opioids and sexual behavior of male rats: involvement of the medial preoptic area.

Local infusion of beta-endorphin (beta-END) into the medial preoptic area (MPOA) dose-dependently impaired the gating of the copulatory response and the execution of the sexual performance of sexually experienced, intact male rats. Local naloxone treatment prevented the impairment of the sexual response by beta-END, but failed to facilitate unimpaired copulation. Local infusion into the MPOA of equimolar doses of alpha-endorphin, dynorphin-A-(1-17) or met-enkephalin were less effective than beta-END. It is suggested that endogenous opioid systems in the MPOA are normally quiescent, and increased activity may be related to disrupted or inhibited male sexual behavior.

Animals↗

Studies of instrumental behavior with sexual reinforcement in male rats (Rattus norvegicus): II. Effects of preoptic area lesions, castration, and testosterone.

We studied effects of lesions to the medial preoptic area (POA), castration, and testosterone replacement on instrumental and unconditioned sexual behavior in male rats. We achieved instrumental measures of sexual motivation by training males to work for an estrous female, presented in an operant chamber under a second-order schedule of reinforcement. POA lesions abolished mounts, intromissions, and ejaculation but did not disrupt instrumental responses, investigation of the female, or abortive mounting attempts. Castration abolished attempts to copulate and also caused a marked decrease in instrumental responses. Testosterone resulted in the prompt reinstatement of instrumental responses and more gradual recovery of unconditioned sexual behavior. We discuss these results in terms of the motivational and performance effects of these neuroendocrine manipulations.

Animals↗

Magnetic resonance imaging of NMDA-induced lesion of the medial preoptic area and changes in sleep, temperature and sex behaviour.

Destruction of the medial preoptic area (mPOA) neurons of rat brain, induced by intracerebral injection of N-methyl D-aspartic acid (NMDA), has been studied by employing the non-invasive Magnetic Resonance Imaging (MRI) technique. Changes in the MRI images are compared and correlated with the functional changes after the mPOA lesion. The progress of the lesion at the injected site has been monitored (using MRI) from 15 min to 1 month after the stereotaxic microinfusion of NMDA (5 micrograms in 0.2 microliter). This study shows that the localised hyperintense (bright) area starts appearing at the mPOA from 3 h after NMDA injection, and the brightness increases progressively for about 2 days. The size and brightness of hyperintense area decrease thereafter. It has not been possible to locate the lesion site after 3 days, using MRI, except in one rat where a vacuole-like area was seen at the NMDA injected site on postmortem histological examination. The reduction in sleep after the mPOA lesion does not show any correlation with the changes in MRI, as it persists throughout the 3 weeks of recording. On the other hand, the initial drastic reduction in male sex behaviour and the increase in body temperature correlated to some extent with the increased brightness in MRI at the site of lesion. The size and location of the hyperintense area, observed during the first 2 days, match with the lesioned area which was histologically identified after 1 month of NMDA administration. Control administration of normal saline into the mPOA did not produce any alteration in the brightness of the MRI image and practically no loss of neurons at the injected site. Though some functional changes have correlation with the alteration in MRI, this cannot be used to interpret the changes in all the physiological parameters. This study also demonstrates that the disappearance of the brightness in MRI should not be taken to indicate a positive prognosis. Though the lesion could not be seen in MRI within 2 hours, its detection after 3-4 h (but within 3 days) after NMDA lesion would give very valuable information for long term studies.

Animals↗

A nitric oxide synthesis inhibitor in the medial preoptic area inhibits copulation and stimulus sensitization in male rats.

Dopamine in the medial preoptic area (MPOA) facilitates copulation in male rats, and nitric oxide (NO) regulates basal and female-stimulated MPOA dopamine release. Microinjection of L-nitro-arginine methyl ester (L-NAME, an NO synthesis inhibitor) into the MPOA blocked copulation in naive rats and impaired copulation in sexually experienced males. In other naive rats, L-NAME or saline was microinjected into the MPOA before each of 7 daily exposures to a receptive female placed over their cage. In a drug-free test on Day 8, copulation by L-NAME-treated rats was similar to that of unexposed controls and was impaired relative to saline-treated males. Therefore, NO in the MPOA is important for copulation and stimulus sensitization in male rats.

Analysis of Variance↗

Opiate receptor ontogeny in the rat medial preoptic area is androgen-dependent.

The relationship of opiate receptors in the medial preoptic area of the hypothalamus (MPOA) to the gonadal steroid hormone environment during development was assessed using regional densitometric analysis of [3H]naloxone binding in autoradiographs prepared using brain sections from 5-day-old male and female rats treated postnatally either with tamoxifen (0.5 mg/kg), flutamide (20 mg/kg), dihydrotestosterone (DHT; 12.5 mg/kg), or sesame oil vehicle. Tamoxifen, a specific estrogen receptor antagonist, did not alter MPOA binding density in either males or females. Flutamide, a specific androgen receptor antagonist, and DHT, a nonaromatizable androgenic compound, altered the MPOA binding density in males and females, respectively. No treatment altered the binding density in several other brain regions. The results suggest that androgen, not estrogen, regulates the differentiation of MPOA opiate receptors. Since the neuronal development in the region is thought to be mediated by estrogen, both hormones probably act concurrently to affect the ontogeny of different parameters in the same brain region.

Androgens↗

NG-monomethyl-L-arginine co-injection attenuates the thermogenic and hyperthermic effects of E2 prostaglandin microinjection into the anterior hypothalamic preoptic area in rats.

Prostaglandin E2 (PGE2) microinjection (25 ng, 250 nl) into the preoptic area of the anterior hypothalamus (POAH) stimulated heat production in brown adipose tissue (BAT) and increased core temperature in urethane-anesthetized rats. These thermogenic and hyperthermic effects were attenuated by co-injection of NG-monomethyl-L-arginine (NMMA, 25 micrograms), a competitive inhibitor of nitric oxide (NO) production from L-arginine. Inclusion of L-arginine (50 micrograms), though not D-arginine (50 micrograms) reversed the inhibitory effect of NMMA (25 micrograms) on intra-POAH PGE2-induced increases in interscapular BAT (IBAT) and core temperatures. Intra-POAH injection of NMMA (25 micrograms) or L-arginine (50 micrograms) alone had no effect on IBAT and core temperatures. The results suggest that the effect on thermoregulation induced by action of PGE2 in the POAH is modulated by a local L-arginine-dependent and NMMA-sensitive NO-generating system.

Animals↗

A quantitative, immunochemical demonstration of the postnatal development of neurotensin in the medial preoptic area of the rat.

The postnatal ontogeny of neurotensin (NT) in the medial preoptic area (MPO) of the rat was quantitatively investigated using immunocytochemical and neurochemical techniques. On postpartum day 0, NT-like immunoreactive neurons could not be identified within the MPO by peroxidase-antiperoxidase histochemistry and, in fact, did not appear until 9 days postpartum. After this age, the number of NT cells in the MPO increased ontogenetically, as did concentrations of NT in MPO extracts measured by radioimmunoassay and enzyme-linked immunosorbent assay. In organotypic MPO explants derived from neonatal rats, NT-like immunoreactivity was detected only after the explants were maintained 14 days in vitro, after which, the number of NT-like immunoreactive somata increased as the time in vitro was extended. These findings indicate that a substantial amount of differentiation occurs postnatally in the MPO both in vivo and in vitro and, more specifically, that the ontogenetic expression of NT may be an intrinsic property of the MPO.

Age Factors↗

Potentiation of pressor response to angiotensin II at the preoptic area in spontaneously hypertensive rat.

Cardiovascular responses to angiotensin II(AII) at the preoptic area (POA) were compared between normotensive Wistar Kyoto rat (WKY) and spontaneously hypertensive rat(SHR) by measuring blood pressure and heart rate under unrestrained, conscious state via a catheter implanted chronically into the abdominal aorta and by injection of drugs into POA through a chronic guide cannula. AII injected into POA at doses of 0.3 ng and 1 ng produced a dose-dependent pressor response, accompanied with a slight decrease of heart rate, in both WKY and SHR. However, in SHR, the pressor response to AII was more than 2 times greater than that in WKY and was quick in onset and lasted about 30 min. When AII in combination with [Sar1, Ile8]-angiotensin II (0.5 microgram), an AII receptor antagonist, were simultaneously administered to POA, the pressor response to AII was strongly inhibited in both WKY and SHR. The results suggest that the pressor response to AII due to its receptor stimulation at POA is markedly potentiated in SHR.

Angiotensin II↗

Immunocytochemical localization of luteinizing hormone-releasing hormone and proopiomelanocortin neurons within the preoptic area and hypothalamus of the bovine brain.

The anatomical locations of proopiomelanocortin (POMC) and luteinizing hormone-releasing hormone (LHRH) neurons were examined in brain tissue from peripubertal female calves and from mature, luteal-phase cows. Biotin-avidin-peroxidase procedures were used for single- and double-labeled immunostaining. LHRH perikarya formed a loosely arranged continuum, extending posteriorly from the diagonal band of Broca, passing through the medial and lateral preoptic areas, and ending within the anterior hypothalamic area. LHRH fibers, apparently directed toward the median eminence, passed (1) posteroventrally in the periventricular area and through the arcuate nucleus, and (2) ventromedially lateral to the arcuate nucleus and medial to the supraoptic nucleus. POMC perikarya were located within and about the arcuate nucleus, some penetrating into the median eminence. Fibers from these POMC perikarya passed ventrally to terminate in the median eminence and to regions ventral to the mammillary nuclei. POMC fibers also projected dorsally and laterally from the arcuate nucleus to other hypothalamic regions as well as anteriorly along the third ventricular wall to the preoptic area, bed nucleus of stria terminalis, and stria terminalis. Less than 6% of the POMC or LHRH processes were in close anatomical apposition to LHRH perikarya and dendrites. Extensive intermingling of LHRH and POMC fibers occurred within zona externa of the median eminence.

Animals↗

Effect on luteinizing hormone secretion of GABA receptor modulation in the medial preoptic area at the time of proestrous luteinizing hormone surge.

Using a bilateral medial preoptic area (MPOA) infusion system in conscious female rats we have investigated the role of the GABA system in this area on the proestrous luteinizing hormone (LH) surge. Fifteen-minute blood samples for LH estimation were taken throughout the afternoon of proestrus from female rats exhibiting 4-day oestrous cycles and implanted at least 2 weeks prior with cerebral guide cannulae. Between 15:00 and 17:00 h rats received an infusion (1 microliter/30 min) of artificial cerebrospinal fluid (n = 7), 10 microM GABA (n = 6) or 10 microM bicuculline methiodide (BMI, n = 6). Animals infused with GABA failed to exhibit an LH surge, while BMI-treated animals displayed an LH surge which was not significantly different to controls. These data show that on the afternoon of proestrus, there are no tonic modulatory actions of the GABA system, acting through the GABAA receptor, on neural elements controlling the LH surge in the MPOA. If, however, GABA levels are elevated in the MPOA at this time then the LH surge is blocked. In conjunction with data from correlative studies showing a decrease in endogenous GABA release prior to the LH surge, we suggest that this fall in activity is an essential component of the LH surge mechanism.

Animals↗