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PubMed1997

Duality of glutamatergic and GABAergic control of pulsatile GnRH secretion by rat hypothalamic explants: I. Effects of antisense oligodeoxynucleotides using explants including or excluding the preoptic area.

Using antisense oligodeoxynucleotides we aimed to study the role of N-methyl-D-aspartate (NMDA) and gamma-aminobutyric acid (GABA) receptors in the mechanism of Gonadotrophin-releasing hormone (GnRH) secretion in vitro. Since GnRH cell bodies are located in the rat preoptic hypothalamus while most GnRH terminals are in the retrochiasmatic hypothalamus, we compared the effects of oligodeoxynucleotides on explants of the whole (preoptic area included) or retrochiasmatic hypothalamus. When GnRH secretion is evoked by muscimol and NMDA, a time-related reduction of GnRH secretion is caused by antisense oligodeoxynucleotides for the beta subunit of the GABAA receptor and the NR2A subunit of the NMDA receptor, respectively. After 6-7 h, binding studies of tritiated ligands show a decrease in GABA- and NMDA-receptor expression. While these antisense effects are observed using whole explants, no such effects are seen using retrochiasmatic explants, indicating that the facilitatory GABAA and NMDA receptors are encoded in the preoptic area. Using several missense oligodeoxynucleotides or antisense for the NR2B and NR2C subunits of the NMDA receptor, the muscimol- and NMDA-evoked release of GnRH is not affected. When spontaneous pulsatile GnRH secretion is studied, the NR2A antisense oligodeoxynucleotides cause an increase of the interpulse interval. This increase is seen using whole but not retrochiasmatic explants. In contrast, the GABAA and NR2C antisense oligodeoxynucleotides result in a reduction of GnRH interpulse interval. Such a reduction is seen using whole as well as retrochiasmatic explants, indicating that the GABAA and NMDA receptors which mediate inhibition of GnRH pulsatility are encoded in the retrochiasmatic hypothalamus. We conclude that NMDA receptors (NR2A subunit) encoded in the preoptic hypothalamus mediate a facilitatory effect on GnRH pulsatility while GABAA and NMDA (NR2C subunit) receptors encoded in the retrochiasmatic hypothalamus mediate an inhibition of GnRH pulsatility. Pulsatile GnRH secretion is affected differently than the agonist-evoked release of GnRH suggesting that the GnRH secretory neurons and the GnRH pulse generator consist of different cellular entities.

Animals↗
PubMed1998

Androgen receptor and mating-induced fos immunoreactivity are co-localized in limbic and midbrain neurons that project to the male rat medial preoptic area.

Two studies were designed to document neuronal colocalization of androgen receptor immunoreactivity and mating-induced Fos immunoreactivity (AR-ir, Fos-ir) in brain of male rats and to examine the extent to which limbic and midbrain neurons that project to the preoptic area are androgen sensitive and activated by mating. Brains from male rats, killed 1 h after ejaculating with receptive females, were examined for Fos-ir and AR-ir and compared with those from control rats not given access to females. PG21 anti-AR and anti-c-fos primary antibodies were visualized by fluorescence microscopy using cyanine-conjugated and fluorescein-conjugated secondary antibodies. In mated males (Expt. 1), Fos-ir and AR-ir were colocalized in neurons of the medial preoptic nucleus (MPN), the dorsal medial amygdala (dMEA), the central tegmental field (CTF), the bed nucleus of the stria terminalis, the anterior hypothalamus, the lateral hypothalamus, and the ventral premamillary nucleus. In Expt. 2, male rats received a unilateral injection of the retrograde tracer FluoroGold (FG) in the preoptic area and four days later were killed after ejaculating with receptive females. Brains were subsequently examined for FG transport, Fos-ir and AR-ir. Fluorogold-containing neurons were present in dMEA and CTF as well as in other hypothalamic and limbic regions known to project to the MPN. In dMEA and CTF, nuclear colocalization of AR-ir and mating-induced Fos-ir was present in a proportion of FG-containing neurons. Sexually relevant information may be carried through the brain by an interconnected network of hormone-sensitive neurons.

Animals↗
PubMed1979

Effects of transecting lateral neural connections of the medial preoptic area on maternal behavior in the rat: nest building, pup retrieval and prolactin secretion.

In lactating female rats bilateral parasagittal cuts transecting the dorsolateral neural connections of the medial preoptic area (MPOA) abolished nest building and retrieving components of maternal behavior, while crouching and nursing were unaffected. While few animals with these cuts were suckled when presented with pups the prolactin secretion response was undiminished when suckling did occur. Whereas previous studies have demonstrated that extensive surgical separation of the medial preoptic area-anterior hypothalamic continuum from the lateral preoptic area-lateral hypothalamus disrupts all aspects of maternal behavior, the present study has determined more specifically the zone of fibers essential for the active components of maternal behavior, i.e., nest building and retrieving. These fibers appear to enter/leave MPO dorsolaterally beneath the crossing of the anterior commissure in the region of the bed nucleus of the stria terminalis. These neural connections are not the same as those that regulate prolactin secretion in the lactating rat.

Animals↗
PubMed1990

Differential acute influence of medial and lateral preoptic areas on sleep-wakefulness in freely moving rats.

The role of preoptic area (POA) in sleep-wakefulness and related EEG changes is well established. Anatomically the area is divided into medial (mPOA) and lateral (IPOA) portions having different physiological functions. Knowledge regarding the differential role, if any, of those two areas in sleep and wakefulness was lacking in the literature. Therefore, an attempt was made in this study, to investigate the same systematically. Experiments were conducted during day and night in freely moving rats. Electrophysiological parameters defining sleep and wakefulness were recorded before and after reversible inactivation of those two areas separately by microinjection of a local anaesthetic, marcain. The responses were opposite in nature depending upon the time, day or night, when the anaesthetic was applied. During the day, anaesthetization induced wakefulness while during the night, sleep was precipitated. However, anaesthetization of both the areas though induced similar qualitative response, the degree of the responses differed significantly. The results suggest that the mPOA is more effective in maintaining tonic sleep while the IPOA is more potent in the maintenance of tonic wakefulness in the normal rats. The finding supports and fits well with the existing knowledge.

Animals↗
PubMed1993

Copulation increases dopamine activity in the medial preoptic area of male rats.

Dopamine (DA) metabolites in microdialysates from the medial preoptic area (MPOA) of male rats increased during copulation. These increases were not observed during eating of a highly palatable food, or if the animal failed to copulate, or if the microdialysis probe was anterior or dorsal to the MPOA. The only two animals with measurable serotonin (5-HT) levels while the female was present were also the only two that either failed to copulate or copulated but failed to ejaculate. These data are consistent with previous evidence for a facilitative role of MPOA DA in the control of male sexual behavior; however, 5-HT activity in the MPOA may impair copulation.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1996

State-dependent changes of extracellular glutamate in the medial preoptic area in freely behaving rats.

Microdialysis technique was applied to the medial preoptic area (mPOA) of the hypothalamus, a crucial site for the regulation of sleep, in order to analyze the interrelationship between the extracellular level of glutamate (Glu) and the vigilance states. Dialysates from the mPOA were sampled at 5-min intervals for 2-h diurnal period with a perfusion rate of 2.0 microliters/min in freely moving rats, whose sleep-waking behaviors were polysomnographically monitored. Extracellular Glu increased during wakefulness, exhibiting a peak at the transition period from wakefulness to non-rapid-eye-movement sleep (NREMS) (18.0% above the average), whereas it decreased during NREMS (9.2% below the average). It is likely that Glu in the mPOA is dynamically involved in the alterations of the vigilance states.

Animals↗
PubMed2003

Preoptic area warming inhibits wake-active neurons in the perifornical lateral hypothalamus.

Activation of the preoptic area (POA) warm sensitive neurons is known to promote non-REM (NREM) sleep and inhibit neuronal discharge in arousal-related brain structures. The perifornical area of the lateral hypothalamus (PF/LH) was recently recognized to be an additional important arousal promoting region. We studied the behavior of PF/LH neurons in rats during the normal sleep-wake cycle and in response to local POA warming. Most PF/LH neurons were wake-active, and exhibited low discharge throughout NREM. Seventy four percent of these wake-active neurons exhibited moderate or strong activation in REM sleep compared to NREM sleep. A substantial group (26%) exhibited very low discharge in REM as well as NREM sleep. Fifty two percent of units in the PF/LH area were responsive to POA warming; 90% of responsive neurons exhibited a significant reduction (-26.47+/-2.16% for 1 degrees C of POA warming) in their discharge rate. The inhibitory effect of POA warming on PF/LH neurons was not associated with EEG slowing. This study supports the hypothesis that sleep induction by POA warm sensitive neurons is mediated through the inhibition of multiple arousal-related structures.

Animals↗
PubMed1989

Oestrogen and noradrenaline modulate endogenous GABA release from slices of the rat medial preoptic area.

Endogenous gamma-aminobutyric acid (GABA) release from the rat medial preoptic area (MPOA) was measured in an in vitro slice technique with sensitive HPLC analysis. Oestrogen is demonstrated to increase GABA activity in the ovariectomised, oestrogen-primed (OVX-EB) rat prior to the luteinising hormone (LH) surge compared with ovariectomised (OVX) animals. Noradrenaline (NA) at a concentration of 10 microM was found to significantly enhance GABA release in response to 30 mM potassium stimulation in both OVX and OVX-EB animals. A significantly greater response to NA was observed in the OVX-EB animal. No effect of NA on basal GABA release was detected. The effects of NA were blocked by the alpha-adrenergic receptor blocker phenoxybenzamine (PB). These data suggest that GABA activity is modulated both by oestrogens and noradrenergic-mediated input in the MPOA.

Animals↗
PubMed1998

Sex differences in estrogenic regulation of preproenkephalin mRNA levels in the medial preoptic area of prepubertal rats.

Opioids have been implicated in sexual differentiation of the brain and in the regulation of reproductive behavior and endocrinology of mammals. Previous studies have indicated that estrogen administration in adults regulates preproenkephalin MRNA levels in several hypothalamic brain nuclei. We have determined preproenkephalin mRNA levels in estrogen-treated juvenile male and female rats to investigate the developmental pattern of estrogenic regulation of enkephalinergic neurons in the medial preoptic area. Rats were treated with estradiol benzoate (20 microgram/kg/day) or oil from day 21 to 23. Sections of the medial preoptic area (mPOA) were studied by in situ hybridization histochemistry at the single cell level and quantified with the assistance of an image analysis system. Our data indicate that males contain higher levels of preproenkephalin mRNA per neuron than females. In addition, our results indicate that estrogen causes an upward shift in the amount of mRNA expressed per cell, females demonstrating a greater response to estrogen than males. An increase in soma cell area following estrogen treatment was observed only in female mPOA enkephalinergic neurons. Taken together, these results indicate a sex difference in total preproenkephalin levels and in estrogenic regulation of preproenkephalin mRNA in the POA of juvenile rats. These results are discussed in relation to the differential role opioids may play in male and female reproductive physiology.

Animals↗
PubMed2002

Allopregnanolone enhancement of GABAergic transmission in rat medial preoptic area neurons.

Gamma-aminobutyric acid (GABA)-mediated transmission in the medial preoptic area (MPOA) of the hypothalamus plays an important role in functions such as sex steroid hormone dynamics and control of body temperature. The action of allopregnanolone, the primary metabolite of progesterone, on GABAergic transmission was investigated by employing patch clamp whole cell recording on acutely dissociated rat MPOA neurons with the functional connection of presynaptic terminals. Allopregnanolone enhanced spontaneous GABA release on the MPOA neurons and induced prolonged decay of miniature GABAergic-inhibitory postsynaptic currents (mIPSCs). The facilitation of GABA release from the presynaptic terminals by allopregnanolone disappeared in Ca2+-free extracellular solution. The presynaptic action of this neurosteroid was also blocked by bumetanide, a blocker of cation-Cl- cotransporters, and by removal of extracellular Na+. The results suggest that allopregnanolone enhances GABAergic transmission at the MPOA neurons by pre- and postsynaptic mechanisms. The enhancement of GABA release by allopregnanolone might require a high Cl- concentration in the presynaptic terminal maintained by Na+-dependent, bumetanide-sensitive mechanisms (e.g., Na+-K+-Cl- cotransporter) and might be mediated by Ca2+ influx into presynaptic terminal.

Animals↗
PubMed1989

Effects of gonadectomy and treatment with gonadal steroids and luteinizing hormone secretion in hypogonadal male and female mice with preoptic area implants.

The ability of male and female hypogonadal (hpg) mice with preoptic area (POA) grafts to show negative feedback was studied. GnRH neurons within POA grafts send axons into the median eminence of the hpg host (HPG/POA), resulting in increased gonadotropin production and gonadal development in the mice that are genetically unable to produce GnRH. The present studies evaluated whether negative feedback, an aspect of normal reproductive function, is present in male and female HPG/POA mice. In normal male mice plasma LH was increased 1 or 5 months after castration and returned to baseline after testosterone propionate treatment. In contrast, no alterations in plasma LH were measured in similarly treated HPG/POA males. HPG/POA female mice were ovariectomized 6 weeks or 3 or 6 months after graft surgery and received sc 17 beta-estradiol (E2) implants 3 months later. Normal mice were studied when 6 weeks old and 8 months old (age-matched to HPG/POA mice ovariectomized 3 months after graft surgery). Further, to determine whether the mice were capable of positive feedback, 1 week after receiving E2 implants, mice in the 6-week and 3-month postgraft surgery groups were challenged with sequential administration of estradiol benzoate and progesterone. The significant increase in plasma LH after ovariectomy or decrease after E2 implant in normal female mice was not present in most HPG/POA female mice. Just 2 of the 24 HPG/POA females studied had increased plasma LH after gonadectomy, and in only 1 of these was plasma LH suppressed by E2 treatment. The ability of an individual HPG/POA mouse to show positive feedback did not predict the ability to show negative feedback, nor did the ability to show negative feedback predict positive feedback capability. Among the mice that failed to respond to ovariectomy with increased LH release were some that had elevated LH in response to steroid challenge or had spontaneously ovulated. On the other hand, neither mouse that had increased LH release after ovariectomy had shown positive feedback to a steroid challenge. Immunocytochemical evaluation revealed GnRH cells within the grafts and GnRH fiber innervation of the host's median eminence, but there was no correlation between numbers of GnRH cells or extent of innervation with the ability to show either negative or positive feedback, nor was the presence of vasoactive intestinal peptide cells within the grafts of predictive value. The failure of negative feedback in most of the HPG/POA mice tested may be due to the failure to establish as yet unidentified but essential afferents to the grafted GnRH cells and/or their axonal processes.

Animals↗
PubMed1990

Angiotensin II antiserum decreases luteinizing hormone-releasing hormone in the median eminence and preoptic area of the rat.

Ovariectomized female rats were sacrificed 3 h after intracerebroventricular microinjection of normal rabbit serum (NRS), specific antiserum against angiotensin II (AB-AII) or atrial natriuretic factor (AB-ANF). AB-AII decreased plasma LH by 50% and LH-RH content by 70% in the median eminence and medial preoptic area, respectively, but did not change plasma FSH when compared to animals which received NRS. There was no difference in these parameters when the animals were injected with AB-ANF or NRS. These results indicate that endogenous AII plays a physiological role in LH release acting directly or indirectly through LH-RH neurons of the median eminence and medial preoptic area.

Angiotensin II↗
PubMed1988

Inhibitory effect of hypothalamic medial preoptic area somatostatin on growth hormone-releasing factor in the rat.

Possible inhibitory effects of somatostatin (SRIF) on GRF were studied by assessing spontaneous GH secretion and GRF content and release in adult male rats depleted of hypothalamic SRIF by anterolateral hypothalamic deafferentation (AHD) or electrolytic lesions in the medial preoptic area (MPO). Plasma GH levels were measured 7 days postoperatively every 20 min in conscious animals with indwelling iv cannulae. Median eminence SRIF was markedly reduced 8 days postoperatively in both AHD and MPO rats, as determined by immunohistochemistry and RIA (P less than 0.01). Although GRF immunoreactivity in the median eminence of AHD and MPO animals appeared well preserved immunocytochemically, hypothalamic GRF content by RIA was significantly decreased at 8 days (P less than 0.01). Spontaneous GH secretion was pulsatile in sham-operated animals. In contrast, basal GH levels in AHD and MPO animals were markedly elevated (P less than 0.01), and secretory pulses were absent. Intravenous injection of specific anti-GRF serum into MPO animals decreased the elevated plasma GH levels (P less than 0.01), indicating increased hypothalamic GRF secretion. GRF release from hypothalamic median eminence-arcuate nucleus complexes in vitro was significantly greater in AHD and MPO animals than in control animals 4 and 8 days postoperatively in response to 30 mM K+ (P less than 0.01), but not under basal conditions. These results suggest that hypothalamic medial preoptic area somatostatinergic neurons play a tonic inhibitory role in the regulation of GRF release and that GH hypersecretion observed after MPO and AHD is attributable to changes in both SRIF and GRF.

Animals↗
PubMed1986

Intermale social aggression: suppression by medial preoptic area lesions.

The intermale social aggressive behavior of male rats cohabiting with a female rat was quantitatively scored weekly in response to the introduction of an unfamiliar intruding male. Resident male rats whose aggressiveness toward an intruder reached a criterion level were subjected to either sham lesions or bilateral lesions in the region of the medial preoptic area. The lesioned rats continued to exhibit levels of piloerection and lateral attack that were not significantly lower than those of sham-lesioned animals. However, the lesioned animals did emit significantly fewer bites and spent significantly less time in the "on-top" position than did sham-lesioned animals. The lesioned animals also displayed significantly less sexual behavior than the sham-lesioned animals but were not different in terms of defensiveness toward the experimenter. It is suggested that bilateral lesions in the region of the medial preoptic area cause a decrease in the intensity of intermale social aggression but do not prevent external stimuli from eliciting the aggression.

Aggression↗
PubMed2001

Partial aortic ligature induces selective long-term c-fos like immunoreactivity in the organum vasculosum of the lamina terminalis, medial preoptic area and choroid plexus in the rat.

Partial aortic ligature causes an increase in water and sodium intake. Circumventricular brain regions are known to be involved in the regulation of these processes. In this work we use c-fos-like immunoreactivity to detect active areas involved in the long-term control of increased water and sodium intake due to partial aortic ligature. A significant increase in water intake was found on the first day after the induction, while natriophilia was observed on the fourth day. c-fos-like immunoreactivity was found selectively in the subfornical organ, the organum vasculosum of the lamina terminalis, the medial preoptic area, and the choroid plexus of the third ventricle. Present results provide further evidence for the involvement of circumventricular organs and the preoptic area in the regulation of hydromineral balance. Moreover, they suggest a maintained and long-term regulation of sodium intake by these same brain areas.

Animals↗
PubMed2002

Increased galanin synapses onto activated gonadotropin-releasing hormone neuronal cell bodies in normal female mice and in functional preoptic area grafts in hypogonadal mice.

Galanin synaptic input onto gonadotropin-releasing hormone (GnRH) neuronal cell bodies was analysed in female mice using the presynaptic vesicle-specific protein, synaptophysin (Syn) as a marker. In the first experiment, forebrain sections from normal ovariectomized ovarian steroid-primed mice exhibiting a surge of luteinizing hormone were processed for immunohistochemical labelling for GnRH, synaptophysin, galanin and Fos. Two representative sections from each brain, one passing through the anterior septum (anterior section) and the other through the organum vasculosum lamina terminalis-preoptic area (posterior section), were analysed under the confocal microscope. None of the GnRH cells analysed in the anterior sections were Fos immunoreactive (IR) or received input from galanin-IR fibres. In contrast, the majority of GnRH cells in the posterior sections analysed were Fos-positive. The number of galanin synapses onto the Fos-positive GnRH cells was significantly higher than that in the Fos-negative cells in this area of the brain, even though the number of Syn-IR appositions was comparable to each other. Transplantation of preoptic area (POA) into the third cerebral ventricle of hypogonadal (HPG) mice corrects deficits in the reproductive system. In the second experiment, synaptic input to GnRH cells was compared between HPG/POA mice with (functional graft) or without (nonfunctional graft) gonadal development. The mean numbers of Syn-IR appositions and galanin synapses per GnRH cell and the proportion of GnRH cells with galanin input were significantly higher in the functional than in the nonfunctional grafts. The results suggest that galanin can act directly on the GnRH cell bodies and may have an important regulatory role on the GnRH system.

Animals↗
PubMed2003

The morphometry of astrocytes in the rostral preoptic area exhibits a diurnal rhythm on proestrus: relationship to the luteinizing hormone surge and effects of age.

The morphometry of astrocytes in the arcuate nucleus exhibits cyclic changes during the estrous cycle leading to dynamic changes in the communication between neurotransmitters and neuropeptides that regulate pituitary hormone secretion. Data suggest that remodeling of direct and/or indirect inputs into GnRH neurons may influence the timing and/or amplitude of the preovulatory LH surge in young rats. We have previously found that aging alters the timing and amplitude of the LH surge. Therefore, the purpose of this study was to focus on the rostral preoptic area where GnRH cell bodies reside. We assessed the possibility that the morphometry of astrocytes in the rostral preoptic area displays time-related and age-dependent changes on proestrus. Our results demonstrate that, in young rats, astrocyte cell surface area decreases between 0800 h and 1200 h, before the initiation of the LH surge. Changes in surface area over the cycle were specific to astrocytes in close apposition to GnRH neurons. In contrast, in middle-aged rats astrocyte surface area was significantly less than in young rats and did not change during the day. These findings suggest that a loss of astrocyte plasticity could lead to the delayed and attenuated LH surge that has been previously observed in middle-aged rats.

Aging↗
PubMed2003

Evidence that thyroid hormones act in the ventromedial preoptic area and the premammillary region of the brain to allow the termination of the breeding season in the ewe.

Thyroid hormones are permissive for various species to enter seasonal anestrus. In the ewe they act centrally to permit the onset of potent estradiol-negative feedback responsible for anestrus, but the specific sites of action are unknown. Therefore, we tested whether T(4) replacement via chronic microimplants in any of five brain areas could reverse the reproductive effects of thyroidectomy. Diffusion of (125)I-T(4) from the microimplant was largely (>98%) limited to a 1.2-mm radius. A marked decline in LH concentration in ovariectomized, estradiol-treated ewes was used as an index for anestrus. In experiment 1, all thyroidectomized (THX) ewes with microimplants in the medial preoptic area, A15 area, and medial basal hypothalamus failed to enter anestrus; instead, LH levels remained elevated, similar to those in untreated THX controls. In ventromedial preoptic area (vmPOA)-microimplanted ewes, only the two animals with the most caudal microimplants entered anestrus, as did thyroid-intact controls and THX ewes receiving icv or sc T(4) replacement. In experiment 2, all vmPOA-treated ewes with similar placements to those effective in experiment 1 along with all ewes microimplanted in the premammillary region entered neuroendocrine anestrus. Thus, the premammillary region and vmPOA are brain sites in which thyroid hormones act to permit the onset of seasonal anestrus.

Anestrus↗