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PubMed1976

Characterization of the androgen receptors in the hypothalamus, preoptic area and brain cortex of the rat.

The specific androgen receptors for testosterone (T) (1) and 5alpha-dihydrotestosterone (DHT) in the cytosol fraction of the hypothalamus, preoptic area and brain cortex of the rat have been characterized using electrophoresis and isoelectric focusing in polyacrylamide gels. After labeling of the cytosol fractions in vivo and in vitro we were able to demonstrate androgen-receptor complexes moving with an electrophoretic mobility (R(f) of 0.5 in 3.25% acrylamide gels containing 0.5% agarose and 10% glycerol. Polyacrylamide gel electrophoresis was used as a quantitative assay for androgen receptors in the tissues. The hypothalamus, preoptic area and brain cortex were found to possess a single class of high affinity binding sites for androgens and the dissociation constants (K(D) were estimated to be 3.4, 4.3 and 2.6 X 10 (-10M) respectively. The binding capacities were 3.7 (hypothalamus), 3.5 (preoptic area) and 1.8 X 10 (-15) (brain cortex) moles of high affinity binding sites per mg protein. Like other androgen-receptor complexes, the testosterone-receptor complexes of the hypothalamus, preoptic area and brain cortex were temperature labile, sulfhydryl dependent and revealed a very slow rate of dissociation at o degrees C (t1/2 greater than 36 hr). The receptors in all the tissues had an isoelectric point of 5.8. The steroid specificity of the cytoplasmic androgen receptors was tested in vitro by the competing efficiency of different unlabeled steroids for (3H)-testosterone binding. In the three tissues in investigation the following order of affinity was found: DHT greater than T greater than Cyproterone acetate greater than progesterone greater than androstenedione greater than 17beta-estradiol. Cortisol did not effect androgen binding significantly. Thus, the physiochemical characteristics of the cytoplasmic androgen receptors of the hypothalamus, preoptic area and brain cortex are very similar, if not identical, to those of the androgen receptors described in the anterior pituitary, ventral prostate, epididymis and testis.

Androgens↗
PubMed1993

Laterality of the gonadotrophin releasing hormone pulse generator in the rat: interaction of neurons located in both preoptic areas as a basic feature.

Though the pivotal importance of pulsatile gonadotrophin releasing hormone (GnRH) release from the hypothalamus for sufficient luteinizing hormone (LH) secretion is well recognized, the mechanisms causing GnRH pulses are still largely unknown. In the rat, the GnRH neurons are concentrated in the preoptic area, which is a bilateral structure of the anterior hypothalamus. This distribution into two distinct brain areas raises the question of whether the GnRH neurons in both preoptic areas are simultaneously or alternatively active and whether an interaction between both areas is required to generate GnRH pulses. We addressed these questions by unilateral manipulations of the activity of GnRH neurons using either electrical lesion or local infusion of gamma-aminobutyric acid (GABA), a neurotransmitter known to inhibit LH release via a hypothalamic action. Acute lesion of one preoptic area caused complete cessation of pulsatile LH release which was restored after 48 h. An acute inhibition of LH release was also observed after unilateral preoptic application of GABA. It is concluded that an interaction of both preoptic areas is mandatory for GnRH pulse generation. However, we suggest that the pacemaker function is not intrinsic to GnRH nor GABA neurons but the cross-talk between these neurons within and between both preoptic areas results in episodic GnRH release.

Animals↗
PubMed1992

Reciprocal connections between the medial preoptic area and the midbrain periaqueductal gray in rat: a WGA-HRP and PHA-L study.

The midbrain periaqueductal gray (PAG) participates in diverse functions such as analgesia, autonomic regulation, sexual behavior, and defense/escape responses. Anatomical studies of the circuits involved in such functions have largely focused on the connections of PAG with the medulla. Projections to PAG from forebrain structures are extensive, but their organization has received little attention. Previous anatomic studies indicate that the medial preoptic area (MPO), involved in a variety of physiological and behavioral functions, is a major source of afferent input to the periaqueductal gray. Here, we have examined the topography of reciprocal connections between these two structures in the rat by using wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) and Phaseolus vulgaris leucoagglutinin (PHA-L). Multiple WGA-HRP injections at several rostrocaudal levels of PAG retrogradely labeled large numbers of neurons in the medial preoptic area; labeled cells were primarily located in the medial preoptic nucleus, the median preoptic nucleus, and the region lateral to the medial preoptic nucleus. The distribution of labeled cells shifted medially to laterally along the rostral to caudal axis of the medial preoptic area. Rostrally, there was selective retrograde labeling in the central and lateral divisions of medial preoptic nucleus, whereas caudally, labeled cells were primarily located only in the lateral subdivision of medial preoptic nucleus. Tracer injections in PAG also produced strong anterograde labeling in MPO. WGA-HRP and PHA-L injections in the medial preoptic area resulted in dense anterograde labeling along the entire rostrocaudal axis of PAG. The terminal labeling in PAG from the medial preoptic area was not uniformly distributed throughout PAG, however. Instead, this projection formed one or two rostrocaudally oriented longitudinal columns that terminated in different subregions of PAG along the entire rostrocaudal axis of this structure. Rostrally, inputs from the medial preoptic area project heavily to dorsomedial PAG, and at mid-PAG levels, the projection becomes distinctly bipartite with two discrete longitudinal terminal columns in dorsomedial and lateral PAG; caudally, the heaviest labeling is in ventrolateral PAG. The projection also exhibited a central to peripheral (radial) gradient; labelled fibers and terminals were heaviest near the aqueduct and much lower in the peripheral parts of PAG. WGA-HRP injections in MPO also produced retrograde labeling of neurons at all rostrocaudal levels of PAG; more neurons were labeled in the rostral than the caudal half of PAG. The majority of labeled cells were located in dorsomedial and ventral/ventrolateral parts of PAG; only a few neurons in the dorsal raphe region appear to project to MPO.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗
PubMed1992

Testosterone regulates substance P within neurons of the medial nucleus of the amygdala, the bed nucleus of the stria terminalis and the medial preoptic area of the male golden hamster.

The medial nucleus of the amygdala, bed nucleus of the stria terminalis, and medial preoptic area appear to mediate steroidal regulation of mating behavior in male rodents. The mechanism of action has not been determined. One way testosterone could enhance neuronal function is by increasing neurotransmitter levels, thus altering neuronal transmission. To assess this hypothesis, we examined the effect of castration and testosterone treatment on substance P levels in the neurons of these three brain regions. Brains from male Syrian hamsters that were (1) gonadally intact, (2) castrated for 13 weeks, or (3) castrated for 9 weeks and treated with testosterone for 4 weeks, were processed for substance P, and the numbers of substance P immunoreactive neurons in the medial nucleus of the amygdala, bed nucleus of the stria terminalis, and medial preoptic area were determined. Castration reduced the number of substance P neurons in the bed nucleus of the stria terminalis and medial preoptic area relative to those in intact hamsters; the number of substance P neurons in these regions was restored by testosterone treatment. Castration did not reduce the number of substance P neurons in the medial nucleus of the amygdala; however, testosterone treatment increased the numbers of these neurons when compared to intacts. Thus, testosterone regulates substance P levels in areas that regulate mating behavior. As substance P enhances male copulatory behavior our results suggest that testosterone may regulate copulatory behavior by enhancing substance P levels in medial nucleus of the amygdala, bed nucleus of the stria terminalis and medial preoptic area.

Amygdala↗
PubMed1995

Warm and cold signals from the preoptic area: which contribute more to the control of shivering in rats?

1. To find out whether the thermosensitive neurones in the preoptic area that control shivering are predominantly warm or cold sensitive, we tested the effects of injecting the excitatory amino acid L-glutamate at various sites in and adjacent to the preoptic area of anaesthetized rats shivering at ambient temperatures of 15-21 degrees C. 2. L-Glutamate injections (0.2 mM in 0.5-1.0 microliter), as well as preoptic warming and electrical stimulation, suppressed shivering, whereas control saline injections had no effect. Effective sites were restricted to the anterior part of the preoptic area, and a tenfold lower concentration of L-glutamate did not influence shivering. 3. Injections of procaine (0.2 M) into the sites where L-glutamate suppressed shivering did not affect strong shivering activity, but facilitated shivering in three out of seven cases when shivering was weak or absent at higher ambient temperatures (25-30 degrees C). 4. L-Glutamate injections, as well as preoptic warming and electrical stimulation, also elicited vasodilatation of the paw skin and the tail. Procaine elicited vasoconstriction when it was applied during vasodilatation induced by local preoptic warming. 5. These results indicate that the contribution of the preoptic area to the control of shivering and vasomotion is influenced more by signals from warm-sensitive neurones than by signals from cold-sensitive neurones.

Animals↗
PubMed1986

A substance P-containing pathway from the hypothalamic ventromedial nucleus to the medial preoptic area of the rat: an immunohistochemical analysis.

The destruction of the hypothalamic ventromedial nucleus, which contains a group of substance P-like immunoreactive neurons, resulted in a marked ipsilateral reduction of these fibers in the medial preoptic area. To test if and to what extent the substance P-like immunoreactive neurons in the ventromedial nucleus project to the medial preoptic area, we applied a sensitive double-labeling method capable of detecting substance P-like immunoreactivity in neurons retrogradely labeled with biotin-wheat germ agglutinin following injection of the tracer in the medial preoptic area. The appearance of many double-labeled cells in the hypothalamic ventromedial nucleus provides strong evidence for the existence of a prominent substance P containing pathway from the ventromedial nucleus to the medial preoptic area. A few doubled-labeled cells were also seen in the lateral hypothalamus, which therefore seems to be an additional source of substance P-like immunoreactive fibers in the medial preoptic area.

Animals↗
PubMed1998

Estrogen implants in the medial preoptic area stimulate maternal behavior in male rats.

The present study investigated whether the medial preoptic area (MPOA) mediates estrogen stimulation of maternal behavior in the male as it does in the female. Previous studies have shown that lesions of the medial preoptic area prevent sensitization of maternal behavior in male rats and that in gonadectomized, hormonally primed males, systemically administered estradiol benzoate stimulates short-latency maternal behavior. These findings are similar to those found in females. In the present study adult males were gonadectomized and hormonally primed with subcutaneously implanted capsules of estradiol (Days 1-16) and progesterone (Days 3-15) and then were stereotaxically implanted bilaterally in the MPOA with implants containing 10% estradiol. Tests with young pups were started 48 h later and continued for 10 days (11 tests). Control groups were implanted in the MPOA with cholesterol or were injected subcutaneously with estradiol benzoate (100 microg/kg). Estradiol implanted males had shorter latencies for maternal behavior (retrieving, crouching, licking pups) than cholesterol implanted males, but their latencies were slightly longer than those of estradiol benzoate injected males. The medial preoptic area, therefore, mediates estrogen stimulation of maternal behavior in males as it does in females.

Animals↗
PubMed2004

Sex differences in the distribution and abundance of androgen receptor mRNA-containing cells in the preoptic area and hypothalamus of the ram and ewe.

Rams and ewes show a negative-feedback response to peripheral treatment with testosterone, with both sexes having a similar degree of suppression in luteinizing hormone (LH) secretion during the breeding season. At least part of the action of testosterone to suppress gonadotropin-releasing hormone/LH secretion is exerted via interaction with an androgen receptor. The distribution of androgen receptor-containing cells in the hypothalamus has been described for the ram, but similar studies have not been performed in the ewe. In the present study, we tested the hypothesis that levels of androgen receptor mRNA expression in the preoptic area and hypothalamus would be similar in rams and ewes. Perfusion-fixed brain tissue was obtained from adult Romney Marsh ewes (luteal phase) and rams during the breeding season (n = 4/sex). Androgen receptor mRNA expression was quantified in hypothalamic sections by in situ hybridization using an (35)S-labelled riboprobe and image analysis. Hybridizing cells were found in the medial preoptic area, bed nucleus of the stria terminalis, anterior hypothalamic area, ventromedial nucleus, arcuate nucleus and premamillary nucleus. The level of androgen receptor mRNA expression was higher in rams than ewes in the rostral preoptic area, caudal preoptic area and rostral portion of the bed nucleus of the stria terminalis, with no sex difference in other regions. The preoptic area and bed nucleus of the stria terminalis are important for reproductive behaviour and the sex differences in androgen receptor mRNA expression at these levels may relate to this. The high level of androgen receptor mRNA expression in the basal hypothalamus, with no sex difference, is consistent with the role of this region in the regulation of gonadotropin secretion.

Animals↗
PubMed2000

Gonadotropin-releasing hormone exhibits circadian rhythm in phase with arginine-vasopressin in co-cultures of the female rat preoptic area and suprachiasmatic nucleus.

To determine whether the suprachiasmatic nucleus can drive a circadian release of gonadotropin-releasing hormone (GnRH) in the preoptic area, we measured the release of GnRH, arginine-vasopressin and vasoactive intestinal polypeptide (VIP) in cocultures of the preoptic area and the suprachiasmatic nucleus at 2-h intervals over a period of 120 h. The release of GnRH in cocultures exhibited a significant circadian rhythm in the presence of oestrogen but not in the absence of oestrogen. The period of the GnRH circadian rhythm was the same as that of the arginine-vasopressin circadian rhythm, and different from the VIP circadian rhythm in each coculture. Furthermore, the peak phase of the GnRH rhythm occurred at the time same as that of the arginine-vasopressin rhythm in each coculture. However, the peak phase of the GnRH rhythm was not always the same as that of the VIP rhythm. Administration of arginine-vasopressin significantly increased GnRH release in single preoptic area cultures in the presence of oestrogen, but VIP did not. The result suggests that, in cocultures of the suprachiasmatic nucleus and the preoptic area, arginine-vasopressin neurones drive the circadian release of GnRH in the presence of oestrogen. We suggest that arginine-vasopressin neurones in the suprachiasmatic nucleus mediate the clock information to GnRH neurones in vivo as well.

Animals↗
PubMed1984

Evidence for a projection from the lateral preoptic area and substantia innominata to the 'mesencephalic locomotor region' in the rat.

A series of anatomical and electrophysiological experiments have been carried out to examine the organization of a direct projection from the substantia innominata and the lateral preoptic area of the hypothalamus, referred to collectively as the subpallidal region, to the pedunculopontine nucleus and adjacent parts of the dorsal midbrain in the adult rat. In the first series of experiments, the retrogradely transported fluorescent tracer SITS, which does not appear to be taken up by fibers-of-passage, was injected into the pedunculopontine nucleus, and the distribution of labeled neurons was plotted in the substantia innominata and lateral preoptic area, as well as in adjacent regions including the medial preoptic area, the bed nucleus of the stria terminalis and parvocellular parts of the paraventricular nucleus. Then, the anterogradely transported lectin PHA-L, which also does not appear to be taken up in effective amounts by fibers-of-passage, was injected into parts of the substantia innominata and lateral preoptic area that project directly to the pedunculopontine nucleus. These experiments demonstrated that fibers from both regions descend through the medial forebrain bundle and give rise in the pedunculopontine nucleus to a terminal field that contains many structures with the appearance of terminal boutons. They also indicated that individual fibers from the subpallidal region innervate both the pedunculopontine nucleus and adjacent parts of the central gray, and that the pathway innervates areas along the length of the medial forebrain bundle on its way to the dorsal midbrain. In a third series of experiments the retrogradely transported fluorescent tracer True Blue was injected into upper thoracic levels of the spinal cord, and it was found that the region of the pedunculopontine nucleus that receives the densest input from the subpallidal region contained many retrogradely labeled neurons on both sides of the brain. And finally, a series of electrophysiological experiments demonstrated that single-pulse stimulation of the substantia innominata and the lateral preoptic area altered the firing rate of a majority of the neurons in and around the pedunculopontine nucleus, and that excitatory and inhibitory responses occurred about equally. These results clearly suggest that the subpallidal region projects directly to the pedunculopontine nucleus and adjacent regions including the central gray and the superior colliculus.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗
PubMedDate not supplied

Efferents of the medial preoptic area in the guinea pig: an autoradiographic study.

Medial preoptic axons were traced into the diagonal band of Broca and septum, particularly lateral septum. Other labeled fibers could be followed dorsally from medial preoptic area injections adjacent to the stria medullaris, and in the periventricular fiber system and the stria terminalis and its bed nucleus. The anterior and medial amygdaloid nuclei were labeled by fibers via the stria terminalis and other arching over the optic tract and through the substantia innominata. The lateral habenula was labeled. Labeled periventricular fibers reached the periventricular nucleus of the thalamus. Descending efferents were traced principally below the fornix and in the adjacent lateral hypothalamus to label the anterior hypothalamus, the tuberal nuclei, and median eminence. Axons of the medial preoptic area joined the medial part of the medial forebrain bundle and distributed to the reticular formation and the central gray of the midbrain and pons. A small amount of contralateral connections were described.

Animals↗
PubMed1997

Differential expression of estrogen receptor mRNA and protein in the female rat preoptic area.

The expression of estrogen receptor (ER) mRNA and ER protein in the medial preoptic area of ovariectomized rat was investigated at both cellular and regional levels using non-isotopic in situ hybridization and immunohistochemistry. ER mRNA was localized in the cytoplasm, while both liganded and unliganded forms of the ER protein were confined to the nucleus. Furthermore, ER mRNA containing cells were evenly distributed throughout the medial preoptic area, showing a homogeneous staining pattern compared to that of ER protein. ER immunoreactive cells were highly distributed in the medial, moderately in the lateral aspect of the medial preoptic area, showing a heterogeneous staining pattern with strongly and weakly labeled cells. These results suggest that ER protein levels are controlled by cellular posttranscriptional mechanisms.

Animals↗
PubMed1995

Co-localization of aromatase enzyme and estrogen receptor immunoreactivity in the preoptic area during reproductive aging.

Immunoreactive aromatase enzyme (AROM-IR) was studied in the preoptic and septal ares of the male Japanese quail brain relative to the age-related decline in endocrine and behavioral components of reproduction. Additional analyses were conducted to determine if the co-localization of AROM-IR and estrogen receptor immunoreactivity (ER-IR) in the medial preoptic area change during aging. Young, sexually active, male quail (6 months of age) were compared to aged sexually active or inactive, male quail (36 months of age). Testis size decreased in old, sexually inactive males, similar to our previous observations. The numbers of AROM-IR neurons in the medial preoptic area (POM) and the lateral septum (LS) decreased significantly with aging and sexual activity. The number of cells that co-localized both AROM-IR and ER-IR did not differ with age. As a consequence of the age-related change in AROM-IR cells, the relative percentage of dual labelled (AROM-IR and ER-IR) and single labelled cells (AROM-IR) increased in aged males. These data provide histochemical evidence that alterations in the aromatase enzyme system in the medial preoptic area may underlie behavioral and endocrine events associated with reproductive aging.

Aging↗
PubMedDate not supplied

Effects of succinylcholine and related substances administered into the medial preoptic area on the local EEG, body temperature, heart rate, galvanic skin resistance and biogenic amines.

Succinylcholine (Sch) which is a cholinergic neuromuscular blocker has been known to occasionally lead to episodes of malignant hyperthermia in swine and humans. In order to find whether it produces any hyperthermic effects through action on medial preoptic area, experiments were carried on by administering intracerebrally the chemical into the medial preoptic area through an in-dwelling cannula-cum-electrode in the free moving rat. The changes in body temperature and the local EEG were studied. For comparison purpose, the effects of carbachol, atropine and phenylephrine were also studied. Further, in the curarized state of no muscular activity, the effect of SCh on the preoptic area was again tested and also the changes in the other autonomic parameters of heart rate and galvanic skin resistance (GSR) were studied. It was observed that SCh given into preoptic area caused a clear hyperthermic effect. The effect was countered by prior administration of atropine into the site. After SCh the local EEG changed into a high amplitude slow wave format. The heart rate was not altered but the GSR increased by two-fold. Carbachol caused a rise in body temperature, heart rate and also GSR. SCh also caused a reduction in noradrenaline content of the hypothalamus by 23% while no change in dopamine and serotonin occurred. Serotonin increased by 28% in the brainstem with no change in the other amines. Septum showed an increase of noradrenaline and dopamine contents by 40% and 25% respectively. Keeping in view the monoaminergic connections and thermoregulatory role of the preoptic area, one may postulate that SCh could inhibit the warm sensors and the controls of the dual sympathetic mechanism which normally leads to an increase of sudomotor activity and a decrease of vasomotor activity, the inhibition resulting in rise of body temperature.

Animals↗
PubMed1986

Autonomic thermoregulation after separation of the preoptic area from the hypothalamus in rats.

To clarify the role of the hypothalamic preoptic area in autonomic thermoregulation, the preoptic area (POA) of rats was disconnected from the rest of the brain-stem by bilateral microknife cuts which spared or included the medial forebrain bundle. The animals' metabolic responses to exogenous norepinephrine (0.5 mg/kg, im) were then measured at ambient temperatures of 25 degrees and 15 degrees C. Oxygen uptake and colonic and tail skin temperatures were also measured at ambient temperatures of 34 degrees, 25 degrees, and 15 degrees C. Finally, the febrile response to a challenge with live Salmonella enteritidis was studied. Except for a slightly higher oxygen uptake at all ambient temperatures in the rats in which the medial forebrain bundle was cut, no differences were found in any of the variables studied between the POA-disconnected and the sham-operated animals. We conclude, therefore, that the POA is not essential for the integration of autonomic thermoregulatory responses in the rat.

Animals↗
PubMed1988

Interrelationship of thermal and sleep-wakefulness changes elicited from the medial preoptic area in rats.

The study investigated the possible interrelationship between changes in sleep-wakefulness and body temperature, primarily induced by manipulation of the noradrenergic system in the medial preoptic area. Saline, norepinephrine, and its alpha- and beta-blockers were injected in the medial preoptic area and in some control areas of rats, during their sleeping and active periods. 5-Hydroxytryptamine was injected in the medial preoptic area in another group of animals. Simultaneous changes in sleep-wakefulness and the body temperature were continuously recorded. Norepinephrine produced hypothermia and arousal, whereas alpha-adrenergic blockers induced hyperthermia and sleep. These changes in body temperature and in sleep-wakefulness did not follow an identical time course. 5-Hydroxytryptamine induced hyperthermia without affecting sleep-wakefulness. It is suggested that there are different neuronal mechanisms in the medial preoptic area that bring about the drug-induced changes in temperature and sleep-wakefulness.

Animals↗
PubMed1987

Effects of hormone deprivation on the karyometric development of the medial and lateral preoptic area of the male mouse. II. Hypothyroidism.

We have analysed the karyometric development of the medial and lateral preoptic area of the hypothalamus in two groups of experimentally hypothyroid male mice, from the 25th to the 160th postnatal day. In one group, animals were treated with propylthiouracil from birth until the day of sacrifice. In the second experimental group animals were treated only until the 35th postnatal day, and thereafter sacrificed at the 45th, 55th, 85th and 160th postnatal day. We compare the results of both groups of animals with a control group of male mice. Hypothyroidism did not alter the nuclear sizes of the medial preoptic area, but in the lateral preoptic area, values were significantly higher in the hypothyroid animals after the 55th day.

Animals↗
PubMed2000

Mu opioid receptor mRNA expression in neuronal nitric oxide synthase-immunopositive preoptic area neurons.

Nitric oxide (NO) as well as beta-endorphin are involved in the neuroendocrine control of gonadotropin-releasing hormone (GnRH) secretion. Recently, morphological and microdialysis experiments have suggested that beta-endorphin may exert an inhibitory influence on NO release in the preoptic area of rat hypothalamus. The present study determines if the mu opioid receptor mRNA is expressed in neuronal NO synthase (nNOS)-immunopositive neurons and if this expression varies among the regions of the basal forebrain being examined. We found, through the use of immunohistochemical and in situ hybridization techniques, that the mu opioid receptor mRNA is expressed in a representative subpopulation of nNOS-immunoreactive neurons in the rat preoptic area. Interestingly, the mu opioid receptor mRNA/nNOS-immunoreactive coexpression is predominant in the rostral and median preoptic area, containing most of GnRH cell bodies. These results strongly suggest that beta-endorphin, via an action through mu opioid receptors, may directly participate in the regulation of NO production in the preoptic area. Our results strengthen the hypothesis that beta-endorphin may participate in GnRH neuronal modulation at the cell body level by regulating NO release from the interneurons of the preoptic area that express nNOS.

Animals↗