Preoptic Suprachiasmatic Nucleus Research Articles

Tyrosine hydroxylase (TH)- and aromatic-L-amino acid decarboxylase (AADC)-immunoreactive neurons of the common marmoset (Callithrix jacchus) brain: an immunohistochemical analysis.

From the perspective of comparative morphology, the distribution of non-monoaminergic neurons in the common marmoset (Callithrix jacchus) was investigated using an immunohistochemical method with specific antibodies to tyrosine hydroxylase (TH) and aromatic-L-amino acid decarboxylase (AADC).TH-immunoreactive (IR) neurons (but not AADC-IR) neurons were observed in the olfactory tubercle, preoptic suprachiasmatic nucleus, periventricular hypothalamic nucleus, arcuate nucleus, paraventricular nucleus, periaqueductal gray matter, medial longitudinal fasciculus, substantia nigra, and nucleus solitaris.In contrast, AADC-IR (but not TH-IR), small, oval and spindle-shaped neurons were sparsely distributed in the following areas: the hypothalamus from the anterior nucleus to the lateral nucleus, the dorsomedial nucleus, the dorsomedial area of the medial mammillary nucleus and the arcuate nucleus; the midbrain, including the stria medullaris and substantia nigra; and the medulla oblongata, including the dorsal area of the nucleus solitaris and the medullary reticular nucleus. The distribution of AADC-IR neurons was not as extensive in the marmoset as it is in rats. However, these neurons were located in the marmoset, but not the rat substantia nigra. Furthermore, AADC-IR neurons that are present in the human striatum were absent in that of the marmoset.The present results indicate that the distribution of non-monoaminergic neurons in the brain of the common marmoset is unique and different from that in humans and rodents.

Organization of projections from the dorsomedial nucleus of the hypothalamus: a PHA-L study in the rat.

The axonal projections of the dorsomedial nucleus of the hypothalamus were investigated by using Phaseolous vulgaris-leucoagglutinin. The main conclusion of this work is that these projections are largely intrahypothalamic, with smaller components directed toward the brainstem and telencephalon. Although the intrahypothalamic pathways are very complex and intermix at various levels, we conclude that dorsomedial nucleus outputs follow three distinct ascending pathways: periventricular, coursing through the hypothalamic periventricular zone; ventral, traveling beneath the medial zone; and lateral, ascending in medial parts of the lateral hypothalamic area. Within the hypothalamus, the most densely innervated areas are the paraventricular nucleus, other dorsal regions of the periventricular zone, the preoptic suprachiasmatic nucleus, and the parastrial nucleus. Other significant terminal fields include the median preoptic, anteroventral periventricular, lateral part of the medial preoptic, and anteroventral preoptic nuclei; and the retrochiasmatic (including perisuprachiasmatic) area. Descending projections follow two pathways that also converge at various levels: a dorsal pathway in the midbrain periventricular system travels through, and primarily innervates, the periaqueductal and pontine gray, and a ventral pathway extends through ventromedial regions of the brainstem. Although sparse, fibers in the later pathway can be traced as far caudally as the nucleus of the solitary tract. The results are discussed relative to the pathways and properties of nearby hypothalamic medial zone nuclei. Dorsomedial nucleus projections are similar to certain other nuclei (e.g., anteroventral periventricular and parastrial) with predominantly intrahypothalamic projections, and different from those arising in the medial zone nuclei (medial preoptic, anterior hypothalamic, ventromedial, and mammillary.

Distribution of NADPH-diaphorase staining and light-induced Fos expression in the rat suprachiasmatic nucleus region supports a role for nitric oxide in the circadian system.

Nitric oxide serves as a messenger molecule in some neuronal systems that use glutamate as a transmitter and it has been shown that glutamate mediates the transmission of photic signals by retinal ganglion cell axons terminating in the hypothalamic suprachiasmatic nucleus, site of the circadian pacemaker in rodents. Recent experiments have demonstrated that pharmacological treatments which block nitric oxide synthesis by nitric oxide synthase prevent glutamate-induced phase shifts of the cell firing rhythm in suprachiasmatic nucleus slice preparation in vitro; similar treatments were found to inhibit light transmission to the suprachiasmatic nucleus as well as light-induced phase shifts in activity rhythms in vivo, implicating nitric oxide in circadian light signalling in vivo. There is limited information, however, about the presence and function of nitric oxide synthase-containing neurons within retinorecipient regions of the rodent suprachiasmatic nucleus. In the present study we used NADPH-diaphorase histochemistry and immunostaining for the nuclear phosphoprotein Fos to assess the co-distribution of nitric oxide synthase-containing neurons and light-responsive cells in the rat suprachiasmatic nucleus region. A strong convergence between NADPH-diaphorase-stained cell bodies and fibres and cells that expressed Fos in response to photic stimulation was noted in the anterior periventricular nucleus, suprachiasmatic preoptic nucleus, retrochiasmatic area, the inter-suprachiasmatic nucleus region, and the dorsal aspect of the optic chiasm, below the suprachiasmatic nucleus. A similar convergence between NADPH-diaphorase-stained fibres and Fos-immunoreactive cells was noted inside the suprachiasmatic nucleus, but the number of NADPH-diaphorase-stained elements found in this region was substantially low compared with that found in retinorecipient regions bordering the nucleus. In many cases both inside and outside the suprachiasmatic nucleus, the Fos-immunoreactive cells appeared to make direct contact with NADPH-diaphorase-stained cells or fibres, but no co-localization of Fos immunoreactivity and NADPH-diaphorase histochemical activity within individual cells was detected. Extensive co-distribution of NADPH-diaphorase-stained cells and fibres and cells that express Fos in response to photic stimulation in the suprachiasmatic nucleus region is in line with the hypothesis that nitric oxide participates in the mechanism mediating circadian light signalling in the suprachiasmatic nucleus. However, lack of co-localization of the two markers to individual cells rules out the possibility that retinorecipient cells in the suprachiasmatic region synthesize and release nitric oxide when photically-activated. Instead, the results support the possibility that photic stimulation triggers nitric oxide synthesis in nitric oxide synthase-containing neurons located near the photically-activated cells.

Autoradiographic localization of [3H]RU 486 and [3H]progesterone in the uterus, pituitary and hypothalamus of the rat

Twenty-one castrated oestrogen-primed Wistar rats, which were 2-months-old, were injected via the jugular vein with 100 mu Ci/100 g body weight of [3H]RU 486 or [3H]progesterone. Some of these received unlabelled compounds for competition studies. Samples of reproductive tract, pituitary and hypothalamus were excised after 15 min. The 4-microns frozen sections were processed for thaw-mounted autoradiography. The exposure time of the autoradiogram was approximately 6 months. After the injection of [3H]RU 486 and [3H]progesterone, the nuclear concentration of radioactivity was most distinct in muscular and stromal cells of the uterus, and the epithelial nuclei of lumina and glands showed weak labelling. Nuclear localization was also observed in muscle cells of the vagina, cervix and oviduct. After injection of [3H]progesterone, the radioactivity was found in the nuclei and cytoplasm of anterior pituitary cells and some cells showed a preferential nuclear concentration of radioactivity. The distribution of [3H]RU 486 in the anterior pituitary was more extensive than that of [3H]progesterone. In the hypothalamus, specific localization of [3H]RU 486 and [3H]progesterone existed in neurones accumulated in the preoptic nucleus, preoptic suprachiasmatic nucleus and the periventricular nucleus. No localization was found in the diaphragm. Pretreatment with RU 486, but not with dexamethasone, reduced the nuclear concentration of radioactivity of [3H]progesterone in the vagina, uterus, oviduct, pituitary and hypothalamus. The nuclear concentration of radioactivity after injection of [3H]RU 486 was also decreased by preinjection with progesterone. The autoradiographic results suggest that RU 486 and progesterone competed for the specific binding site (possibly a progesterone receptor) in the target cells at the levels of the uterus, pituitary and hypothalamus in vivo.

Autoradiographic localization of [3H]RU 486 and [3H]progesterone in the uterus, pituitary and hypothalamus of the rat

Twenty-one castrated oestrogen-primed Wistar rats, which were 2-months-old, were injected via the jugular vein with 100 microCi/100 g body weight of [3H]RU 486 or [3H]progesterone. Some of these received unlabelled compounds for competition studies. Samples of reproductive tract, pituitary and hypothalamus were excised after 15 min. The 4-micron frozen sections were processed for thaw-mounted autoradiography. The exposure time of the autoradiogram was approximately 6 months. After the injection of [3H]RU 486 and [3H]progesterone, the nuclear concentration of radioactivity was most distinct in muscular and stromal cells of the uterus, and the epithelial nuclei of lumina and glands showed weak labelling. Nuclear localization was also observed in muscle cells of the vagina, cervix and oviduct. After injection of [3H]progesterone, the radioactivity was found in the nuclei and cytoplasm of anterior pituitary cells and some cells showed a preferential nuclear concentration of radioactivity. The distribution of [3H]RU 486 in the anterior pituitary was more extensive than that of [3H]progesterone. In the hypothalamus, specific localization of [3H]RU 486 and [3H]progesterone existed in neurones accumulated in the preoptic nucleus, preoptic suprachiasmatic nucleus and the periventricular nucleus. No localization was found in the diaphragm. Pretreatment with RU 486, but not with dexamethasone, reduced the nuclear concentration of radioactivity of [3H]progesterone in the vagina, uterus, oviduct, pituitary and hypothalamus. The nuclear concentration of radioactivity after injection of [3H]RU 486 was also decreased by preinjection with progesterone. The autoradiographic results suggest that RU 486 and progesterone competed for the specific binding site (possibly a progesterone receptor) in the target cells at the levels of the uterus, pituitary and hypothalamus in vivo.

Hormonal control of the development and regulation of tyrosine hydroxylase expression within a sexually dimorphic population of dopaminergic cells in the hypothalamus

In situ hybridization histochemistry was used to examine the development and regulation of tyrosine hydroxylase (TH) mRNA within the sexually dimorphic population of dopaminergic cells in the anteroventral periventricular nucleus (AVPv) of the hypothalamus. The AVPv contains over 3 times as many TH mRNA-containing cells in female rats, compared with males. This sexual dimorphism appears to be dependent on perinatal levels of gonadal steroids since orchidectomy of newborn males increased, and treatment of newborn females with testosterone decreased, the number of TH mRNA-containing cells detected within the AVPv. In addition, circulating gonadal steroids appear to downregulate TH expression within these cells in both adult male and female rats. In adult male animals, gonadectomy increased the number of TH mRNA cells in the AVPv within 7 days. Similarly, estradiol treatment prevented the increase in the number of TH mRNA-containing cells within the AVPv seen in ovariectomized female rats. No sexual differences were detected in the number of TH mRNA-containing cells within the suprachiasmatic preoptic nucleus, located just ventral to the AVPv. These findings indicate that perinatal gonadal steroids influence the number of cells within the AVPv that express TH in detectable amounts by determining the number of cells that are capable of producing sufficient quantities of TH message, as opposed to sex-specific alterations in the post-translational mechanisms. In the adult, circulating gonadal steroids appear to downregulate TH expression within these cells suggesting that testosterone and/or estrogen may exert a sustained influence on the biosynthetic activity of this sexually dimorphic population of dopaminergic cells.

Propranolol and methylatropine antagonize the cardiovascular effects produced by microinjection of the TRH analog MK-771 into the preoptic suprachiasmatic nucleus.

Thyrotropin-releasing hormone (TRH) has been shown to increase heart rate as well as blood pressure when administered into rat brain. The present study investigated the mechanism by which the TRH analog MK-771 produces these effects when injected into the preoptic suprachiasmatic nucleus (POSC). MK-771, at a dose of 125 pmol (50 ng), produced significant increases in both heart rate and blood pressure. These effects occurred within 5 minutes of microinjection and lasted approximately 20-30 minutes. Pretreatment with either the beta-adrenergic antagonist propranolol or the muscarinic antagonist methylatropine, administered into the POSC, significantly altered the response produced by MK-771. Propranolol, at a dose of 7 nmol, and methylatropine at a dose of 0.5 nmol, significantly inhibited the tachycardia produced by MK-771. In addition, methylatropine, at a dose of 0.5 nmol, significantly reduced the increase in diastolic pressure produced by the TRH agonist. These results are consistent with the idea that TRH agonists, when administered centrally, produce cardiovascular alterations through the autonomic nervous system.

The patterns of [ 14C]2-deoxyglucose uptake in female rat brain produced by electrical stimulation of hypothalamic and limbic brain areas

The aim of this study was to investigate the pattern of [ 14C]2-deoxyglucose uptake in anaesthetized rat brain produced by electrical stimulation of brain areas implicated, by previous electrical stimulation studies, in the neural control of pituitary hormone and especially gonadotrophin secretion. Stimulation of the median eminence led to a significant increase in the relative metabolic activities of the arcuate, ventromedial hypothalamic, supraoptic and paraventricular nuclei and the preoptic area. Stimulation of the suprachiasmatic or paraventricular nuclei or the medial preoptic area, anterior hypothalamic area, the dorsal or ventral hippocampus or amygdala led to an increase in the relative metabolic activity of many brain regions known to have direct connections with these areas, but in addition produced increases in the relative metabolic activity of areas which have secondary connections. Hippocampal stimulation confirmed previous neuroanatomical findings of major intrinsic functional connections between different fields of the ipsilateral and contralateral hippocampus. Stimulation of the amygdala, unexpectedly, did not change the relative metabolic activity of the arcuate nucleus and medial preoptic area which have neuroanatomical connections with the amygdala. Similarly, stimulation of the medial preoptic area did not change significantly the relative metabolic activity of the mamillary body and dorsomedial thalamic area. The effect of preoptic area stimulation on the relative metabolic activity of several brain regions was changed by ovariectomy and by injection of oestradiol benzoate. Stimulation of the preoptic area and suprachiasmatic nuclei, but not the anterior hypothalamic area or other brain regions, increased significantly the plasma concentrations of luteinizing hormone. These results show that (i) electrical stimulation of brain areas concerned with the control of gonadotrophin and other pituitary hormone secretion changes the metabolic activity of nuclei and neural pathways extrinsic as well as intrinsic to the hypothalamic-pituitary system, (ii) the [ 14C]2-deoxyglucose method can detect changes in antidromic as well as orthodromic activity and in multisynaptic pathways, (iii) neuroanatomical pathways are not always activated metabolically by electrical stimulation, (iv) the preoptic-suprachiasmatic nucleus gonadotrophin control system is discrete and is little affected by increased metabolic activity of the hypothalamus produced by stimulation of the anterior hypothalamic area or other brain areas.

Diurnal rhythmicity and hypothalamic deficits in glucose utilization in aged ovariectomized rats

Aging is associated with a loss of cyclic gonadotropin release in female animals. This deficit may reflect dampened circadian rhythmicity of neuroendocrine events and/or altered function in hypothalamic nuclei important to regulation of cyclic female reproduction. The purpose of this study was to determine if diurnal periodicity and glucose metabolism in the hypothalamus are altered with age and whether such changes could help to explain the age-related deficits in gonadotropin release. Young (3-4-month-old) and old (18-21-month-old) rats were ovariectomized and subjected to the 2-deoxy-D-1-14C-glucose technique to measure rates of cerebral glucose utilization (GU), an index of neural function (Sokoloff et al., 1977) in various brain areas and in the pineal gland. We measured GU during the light (1400 hours) and the dark (2200 hours) in 17 anatomical regions including the following hypothalamic areas: medial preoptic nucleus, suprachiasmatic preoptic nucleus, suprachiasmatic nucleus, paraventricular nucleus, arcuate nucleus, and median eminence. Serum concentrations of luteinizing hormone (LH) and prolactin were measured in the same rats to determine the effect of age on both of these hormones. Diurnal periodicity of GU was observed in the suprachiasmatic nucleus and the pineal gland in young and old rats. Although there was no age difference in GU of the pineal gland, GU was reduced during the light and dark in the suprachiasmatic nucleus and all other hypothalamic areas examined except the suprachiasmatic preoptic nucleus and the median eminence. Ovariectomy induced an attenuated increase in concentrations of LH in old, compared to young rats.(ABSTRACT TRUNCATED AT 250 WORDS)

The preoptic-suprachiasmatic nuclei though morphologically heterogeneous are equally affected by streptozotocin diabetes

Pituitary and gonadal disorders consistent with abnormal LHRH and LH secretion occur in streptozotocin-diabetic rats. A key role in the synthesis and regulation of LHRH and in the phasic LH release is played by the preoptic-suprachiasmatic region which is mainly formed by the medial preoptic area, the sexually dimorphic nucleus of the medial preoptic area, and the suprachiasmatic nucleus. Therefore we have studied this region by morphology and morphometry in normal and streptozotocin-diabetic rats. In normal animals, the neurons of the above mentioned nuclei were morphologically and morphometrically dissimilar. Independent of their localization, reduced cytoplasmic and nuclear areas were observed in the neurons of diabetic animals. These lesions are consistent with hypotrophied neurons. Consequently, diabetes may impair both synthesis and regulation of LHRH and may therefore account for pituitary disorders, testicular atrophy, and lacking preovulatory LH peaks. The structural differences of the neurons of the three nuclei in normal animals underline their different physiological role. Yet, the similarity of the changes found in all three nuclei suggests a generalized hypofunction of the whole preoptic-suprachiasmatic region under diabetic condition.

The distribution of neurotransmitter-specific cells and fibers in the anteroventral periventricular nucleus: Implications for the control of gonadotropin secretion in the rat

The anteroventral periventricular nucleus (AVPv), which lies in the periventricular zone of the preoptic region, is critical for normal phasic gonadotropin secretion since lesions of this nucleus abolish the progesterone-induced surge of luteinizing hormone secretion from the anterior pituitary, block ovulation, and induce persistent vaginal estrus in female rats. However, very little is known about the neurotransmitter-specific pathways associated with this nucleus. In the present study we evaluated the distribution of biochemically specific cells and fibers within the AVPv and adjacent regions by using an indirect immunohistochemical method with antisera to serotonin (5-HT), dopamine β-hydroxylase (DBH), tyrosine hydroxylase (TH), neuropeptide Y (NPY), cholecystokinin-8 (CCK), vasoactive intestinal polypeptide (VIP), substance P (SP), neurotensin (NT), corticotropin-releasing factor (CRF), luteotropin-releasing hormone (LRH), somatostatin (SS), thyrotropin-releasing hormone (TRH), oxytocin (OXY), vasopressin (OXY), adrenocorticotropic hormone (ACTH 1−24), alpha-melanocyte-stimulating hormone (α-MSH), leucine-enkephalin (L-ENK), and calcitonin gene-related peptide (CGRP). Our findings indicate that both cells and fibers containing these putative neurotransmitters are differentially distributed in and around the AVPv in accordance with the cytoarchitectonic organization of this part of the preoptic region. The AVPv itself appears to receive strong inputs from SP-, VAS-, CCK-, and SS-containing pathways, whereas the highest densities of L-ENK-, NT-, 5-HT-, NPY-, and DBH-immunoreactive fibers were found in the cell-sparse zone just lateral to the AVPv. The suprachiasmatic preoptic nucleus (PSCh), a small group of cells located ventral to the AVPv just dorsal to the optic chiasm, contained high densities of α-MSH- and ACTH-immunoreactive fibers, as well as substantial numbers of fibers containing catecholamines or NPY. In contrast, a dense plexus of VAS-stained fibers was distributed fairly evenly throughout the AVPv and PSCh. Numerous L-ENK-immunoreactive cell bodies, and moderate numbers of CCK-, NT-, and CRF-stained cell bodies were found in the AVPv. The PSCh contained many TH-stained cells (presumably dopaminergic), in addition to a moderate number of CCK-containing cell bodies, while a high density of NT- and CRF-stained cells were found in the cell-sparse zone lateral to the AVPv, in addition to several CCK-, SP-, VIP-, and TH-containing cells. These results are consistent with the view that the AVPv may be a nodal point in which a variety of putative neurotransmitter-specific pathways influence the neural control of gonadotropin-release, and perhaps other visceral functions as well.

Electrophysiological test of an amphiphilic β-structure in LHRH action

Micropipettes were used to record electrical activity from single neurons in hypothalamic tissue slices, in the hypothalamic arcuate nucleus (ARC), and in the periventricular and suprachiasmatic preoptic nuclei (POA). Responses were measured following in vitro application of luteinizing hormone releasing hormone (LHRH), and two analogues: LHRH models 1 and 3. Model 1 (pyroGlu-His-Trp-Ser-Phe-Thr-Ile-Lys-Ile-ThrNH 2) had amino acid substitutions in residues 5–10 designed to form an amphiphilic β-strand structure. Model 3 (pyroGlu-His-Trp-Ser-Phe-Gly-Ile-Lys-Pro-SerNH 2) was also designed to possess amphiphilic characteristics, but also more closely to resemble the native peptide. Electrical recording results showed that LHRH was able both to excite or inhibit different hypothalamic neurons, and that it was more effective in the preoptic area than in the arcuate nucleus. Responses to LHRH model 3 were strongly correlated with responses to LHRH, in their occurrence and their direction, for both of the brain regions studied. Moreover, LHRH models 1 and 3 also retained the neuromodulatory effects of LHRH on cellular responses to norepinephrine and serotonin. Thus, LHRH analogues, designed to possess an amphiphilic β-structure, preserved some of the properties of LHRH when tested electrophysiologically in the central nervous system.

Cardiovascular Effects Produced by Microinjection of Atrial Natriuretic Peptide (ANP) into the Preoptic Suprachiasmatic Nucleus (POSC) of the Rat

Cardiovascular Effects Produced by Microinjection of Atrial Natriuretic Peptide (ANP) into the Preoptic Suprachiasmatic Nucleus (POSC) of the Rat

Electron-microscopic cytochemistry of the catecholaminergic innervation of TRH neurons in the rat hypothalamus.

The catecholaminergic innervation of thyrotropin-releasing hormone (TRH) neurons was examined by use of a combined method of 5-hydroxydopamine (5-OHDA) uptake or autoradiography after intraventricular injection of 3H-noradrenaline (3H-NA) and immunocytochemistry for TRH in the same tissue sections at the electron-microscopic level. TRH-like immunoreactive nerve cell bodies were distributed abundantly in the parvocellular part of the paraventricular nucleus (PVN), in the suprachiasmatic preoptic nucleus and in the dorsomedial nucleus of the rat hypothalamus. In the PVN, a large number of immunonegative axon terminals were found to make synaptic contact with TRH-like immunoreactive cell bodies and fibers. In the combined autoradiography or 5-OHDA labeling with immunocytochemistry, axon terminals labeled with 3H-NA or 5-OHDA were found to form synaptic contacts with the TRH immunoreactive nerve cell bodies and fibers. These findings suggest that catecholamine-containing neurons, probably noradrenergic, may innervate TRH neurons to regulate TRH secretion via synapses with other unknown neurons in the rat PVN.

Medianosomes as integrative units in the external layer of the median eminence. Studies on grf/catecholamine and somatostatin/catecholamine interactions in the hypothalamus of the male rat☆

Somatostatin/catecholamine as well as growth hormone releasing factor/catecholamine interactions have been characterized in the hypothalamus and the preoptic area using morphometrical and quantitative histofluorimetrical analyses. The combined results of the present and previous studies have led us to put forward the medianosome concept. The medianosome is defined as an integrative unit, which consists of well defined aggregates of transmitter identified nerve terminals interacting with one another in the external layer of the median eminence. Our present data indicate the existence of putative medianosomes consisting predominantly of growth hormone releasing factor nerve terminals costoring dopamine as well as of somatostatin and dopamine nerve terminals, which interact locally to control growth hormone secretion. A complementary control of growth hormone secretion may be exerted by noradrenaline mechanisms in the subependymal layer, in the ventral zone and/or in the suprachiasmatic preoptic nucleus. However, further analyses in view of the differential effects seen with the present doses of rat hypothalamic and human pancreatic growth hormone releasing factor have to be done. The results also indicate the possible existence of growth hormone releasing factor receptors in the median eminence which may participate in the feedback control of the growth hormone releasing factor immunoreactive neurons in the ventral zone of the hypothalamus.

Increases in heart rate and blood pressure produced by microinjections of atrial natriuretic factor into the AV3V region of rat brain

Recent studies have provided evidence for the dense localization of atrial natriuretic factor (ANF) in the anteroventral third ventricle (AV3V) region of the rat brain. This area is currently thought to be involved in the regulation of blood pressure and fluid and electrolyte balance. To investigate whether ANF may play a role in central cardiovascular regulation, the effects of microinjection of ANF into the preoptic suprachiasmatic nucleus (POSC), which is located in the AV3V region of the brain, were examined in the present study. Low doses of ANF (2–4 pmol) produced modest elevations in systolic and diastolic pressures, approximately 10–14%, and a small rise in HR of roughly 7%. Higher doses of ANF (20–40 pmol) produced significant increases in systolic (15–19%), mean arterial (12–14%) and pulse (25–36%) pressures. In addition, much larger increases in HR, approximately 20%, were produced by these higher doses of ANF. The onset of effects produced by ANF on BP and HR was seen 15–45 min after injection. Peak effects were usually observed approximately 60–150 min after onset, and the duration of the effect was 2–4 hours, after which time values usually returned to baseline. These studies indicate that ANF produces significant increases in BP and HR when injected at pmol doses into the POSC, and lends support to the idea that this peptide may play an important role in central cardiovascular regulatory mechanisms.

Effects of estradiol and prolactin on incertohypothalamic dopaminergic neurons in the male rat

The activity of incertohypothalamic, tuberoinfundibular, mesolimbic and nigrostriatal dopamine (DA) neurons was estimated by measuring the rate of turnover of DA (decline of DA after α-methyltyrosine) in brain regions containing the cell bodies and terminals of these neurons. Estradiol benzoate (25 μg/kg/day × 3; s.c.), markedly increased the concentration of prolactin in plasma and increased the rate of turnover of DA in median eminence, but failed to alter DA neuronal activity in the striatum or nucleus accumbens. An intracerebroventricular injection of prolactin (10 μg/rat, 12 h) mimicked the effect of estradiol on the turnover of DA in the median eminence. Within the incertohypothalamic DA system, both estradiol and prolactin increased DA turnover in the dorsomedial nucleus but decreased DA turnover in the rostral periventricular, medial preoptic and preoptico-suprachiasmatic nuclei. Hypophysectomy decreased the tonic activity of tuberoinfundibular DA neurons, but failed to alter the activity of incertohypothalamic DA neurons. The stimulatory effects of estrogen on DA turnover in the median eminence and dorsomedial nucleus, and the inhibitory effects of this hormone on rostral hypothalamus DA neurons were blocked by hypophysectomy. Taken together these results suggest that estradiol-induced changes in intact male rats are mediated by prolactin. Estradiol increased incertohypothalamic DA neuronal activity in the medial preoptic and preoptic-suprachiasmatic nuclei of hypophysectomized rats suggesting that in the absence of circulating prolactin, estradiol has a direct stimulatory effect on incertohypothalamic DA neurons in the rostral, periventricular hypothalamus. Thus, while incertohypothalamic DA neurons resemble the major ascending extrahypothalamic DA neurons in their responsiveness to DA agonists and antagonists, they differ from the mesolimbic and nigrostriatal DA neurons in that they are also responsive to estrogen and prolactin, and in this regard resemble tuberoinfundibular DA neurons.

Cardiovascular effects produced by injections of thyrotropin-releasing hormone in specific preoptic and hypothalamic nuclei in the rat

Microinjection of 1.4 pmol TRH (0.5 ng; 50–150 nl) into both the preoptic suprachiasmatic nucleus (pos) and the A6800–7000 region of the medial preoptic nucleus (pom) produced increases in blood pressure and heart rate of 7% and 19%, respectively; heart rate responses in these two areas were higher than those occurring in other areas tested. TRH induced a significant increase in blood pressure and heart rate in the posterior hypothalamic nucleus (nhp) and increased heart rate only in the anterior (nha) and dorsomedial (ndm) hypothalamic nuclei. A small decrease in both blood pressure and heart rate resulted with TRH injections in the A7050–7400 region of the pom. No changes in respiratory rate or rectal temperature were observed at any site with this dose of TRH. Preliminary studies into the mechanism of the cardiovascular actions of TRH suggested that inhibition of the parasympathetic nerves to the heart make a partial contribution to the TRH-induced heart rate increase in the pos and that adrenal catecholamine release mediates the TRH response in the nhp. Neither methylatropine pretreatment nor adrenalectomy prevented the response to TRH injected into the nha, suggesting that activation of the cardiac sympathetic nerves may mediate TRH actions in this region. In the ndm, neither methylatropine nor adrenalectomy prevented the response to TRH; however, there was a tendency for the response to be less after methylatropine. Therefore, both inhibition of the parasympathetic and activation of the sympathetic nervous systems may contribute to the response observed, but no adrenal involvement could be demonstrated. Discrete injections of 0.8 nmol TRH produced increases in heart rate and blood pressure in all preoptic and hypothalamic nuclei tested with accompanying changes in respiratory rate and rectal temperature in some areas. Lateral cerebral ventricle injections of as little as 2.8 pmol TRH produced increases in blood pressure and heart rate; cardiovascular responses to higher doses (0.8–22 nmol) in the ventricle were often accompanied by arousal, piloerection, “wet dog” shakes and changes in respiratory rate and rectal temperature. Previous immunohistochemical demonstration of nerve cells and fibers in the preoptic-hypothalamic area and the present finding of specific sites responsive to low dose TRH injections (1.4 pmol) both support a physiological role for this peptide in central control of the cardiovascular system.

Cardiovascular effects of discrete intrahypothalamic and preoptic injections of Bradykinin

Blood pressure and heart rate were monitored during discrete injections of bradykinin (5 nmol; 100–300 nl) in the hypothalamus and preoptic area of halothane anesthetized rats. In the paraventricular nucleus, bradykinin produced bradycardia without effecting blood pressure. The decrease in heart rate was abolished by pretreatment with methylatropine (IP), suggesting that the parasympathetic nervous system mediates this response. In contrast, in the dorsomedial and posterior hypothalamic nuclei, bradykinin increased both heart rate and blood pressure; methylatropine pretreatment (but not adrenalectomy) blocked these responses, suggesting that inhibition of the parasympathetic nervous system is responsible for the actions of bradykinin in these nuclei. In the preoptic suprachiasmatic nucleus, bradykinin produced an increase in heart rate only, which was attenuated by either methylatropine or adrenalectomy, indicating that both inhibition of the parasympathetic nervous system and adrenal catecholamine release contribute to the actions of bradykinin at this site. The increase in heart rate observed with bradykinin in the medial preoptic and anterior hypothalamic (A6400-6001 region) nuclei was not effected by either methylatropine or adrenalectomy, therefore activation of the sympathetic nervous system may be involved in responses in these regions. Finally, a 5 nmol dose of bradykinin potentiating factor (converting enzyme inhibititor; CEI) had effects similar to bradykinin when injected into the posterior hypothalamus, but no effect at any other brain site. CEI administration into brain sites 15 min prior to bradykinin injections failed to alter the bradykinin response. In summary, the central cardiovascular responses to bradykinin depend upon the specific site of injection and these sites correspond with the localization of bradykinin-like immunoreactivity previously reported by others.

Role of the ventrolateral part of the medulla oblongata in the control of gonadotropin secretion. I. Electrophysiological studies on the neural features between the limbic-preoptic structures and the ventrolateral part of the medulla oblongata in the female rat

Electrophysiological studies were undertaken in order to investigate the neural features between the preoptic suprachiasmatic nucleus (POSC) and the ventrolateral part of the medulla oblongata (VLMO) and between the bed nucleus of stria terminalis (BST) and VLMO by eliciting antidromic responses and performing microionophoresis of estradiol. Unit activity of neurons in the VLMO was recorded in ovariectomized estradiol benzoate-primed Wistar female rats under urethane anesthesia (1.2g/kg body weight). Antidromic activations were performed by applying negative square wave pulses (intensity 1.0--1.5mA, duration 0.1msec, frequency 0.6Hz) which were delivered through a bipolar concentric stainless steel electrode to the POSC or the BST. In total, 116 neurons in the POSC and 118 neurons in the BST with ascending projection were identified in the VLMO. Both in the POSC and the BST, two types of antidromic spike potentials were distinguished on the basis of their waveforms. One type ("fast spikes") was characterized by a sharp and smooth rising phase. The other ("slow spikes") had a notch in the rising phase. In the POSC, mean antidromic spike latency for the fast spikes was 9.8msec, while the value for the slow spikes was 30.4msec and in the BST, mean antidromic spike latency for the fast spikes was 10.2msec, while the value for the slow spikes was 31.5msec. In the POSC, ionophoretic injection of estradiol hemisuccinate was accomplished on 57 of the 116 antidromically identified cells, of which 32 showed slow responses and 25 responded with fast spikes. In cells with slow spikes, estradiol hemisuccinate facilitated (n = 14) or suppressed (n = 5) their generation of action potentials. None of the cells with fast responses changed their activity in response to estradiol hemisuccinate. In the BST, ionophoretic injection of estradiol hemisuccinate was accomplished on 51 of 118 antidromically identified cells, of which 30 showed slow responses and 21 responded with fast spikes. In cells with slow spikes, estradiol hemisuccinate facilitated (n = 9) or suppressed (n = 8) their generation of action potentials. None of the cells with fast responses changed their activity in response to estradiol hemisuccinate. In addition, POSC or BST unit responses to the VLMO stimulation (intensity 1.0--1.5mA, duration 0.1msec, frequency 0.6Hz) was examined. Unit firing activity in the POSC or BST, elicited by the VLMO stimulation, was much more enhanced in proestrous rats than in diestrous rats. After the injection of phenoxybenzamine, an alpha-adrenergic receptor blocker, the rate of facilitatory responses in the POSC or in the BST neurons was significantly decreased in proestrous rats.(ABSTRACT TRUNCATED AT 400 WORDS)