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PubMed1976

Organization and morphologies of acetylcholinesterase-containing neurons in the thalamus and hypothalamus of the rat.

The distribution and morphologies of neurons containing acetyl cholinesterase (AChE, EC 3.1.1.7) in the thalamus and hypothalamus of the rat were studied by means of a pharmaco-histochemical technique involving staining for AChE (Karnovsky-Roots' procedure) at various times after administration of bis-(u-methylethyl)phosphorofluoridate. This method enables visualization of individual AChE-containing neuronal somata and their processes to a degree not possible with other protocols for the enyzme. The strongest AChE activity occurring at the level of the thalamus is found within the small, round to oval, somata of nucleus anterior dorsalis. Most of the intralaminar nuclei, as well as nucleus reticularis, are composed of medium-sized multipolar neurons displaying moderate to strong AChE activity. Moderately stained AChE neurons are also found in pars ventralis of nucleus geniculatus lateralis and in pars lateralis of nucleus habenularis. Most of the neurons of the lateral and posterior thalamic territories, however, are nearly devoid of AChE. At the level of the hypothalamus, the neurons of nuclei supraoptics and paraventricularis show strong AChE activity. The AChE neurons of nucleus supraopticus are surrounded by numerous AChE-containing processes of some large lateral preoptic area neurons that stain intensely for the enzyme. Numerous intensely stained AChE perikarya occur in the lateral, dorsal, and supra-mammillary hypothalmic areas. These neurons often possess several AChE-containing processes. Nuclei arcuatus and ventomedialis consist mainly of neurons displaying a weak to moderate intensity of AChE staining. At the level of the mamillary bodies most neurons show moderate AChE activity except the neuronal somata of nucleus mammillaris lateralis which stain very strongly for the enzyme.

Acetylcholinesterase↗

PubMed1977

Abnormal development of the suprachiasmatic nuclei of the hypothalamus in a strain of genetically anophthalmic mice.

In the anophthalmic mutant of the mouse the optic primordia are "genetically enucleated" well before the usual emergence of retinal ganglion cell axons (Silver and Hughes, '74). In eyeless animals, a portion of the mediobasal hypothalamus and one of its constituent nuclear pairs, nucleus suprachiasmaticus (SCN), were markedly abnormal in the embryo and adult. It has been reported that the ventral portion of the SCN receives a substantial, direct retinal innervation (Moore and Lenn, '72) and that these nuclei may mediate several light-induced hormonal and behavioral circadian rhythms (Stetson and Whitmyre, '76). During day 13 of mutant embryogenesis, just prior to the time of optic chiasm formation in normal animals, a large portion of ependyma and adjacent brain tissue herniated into the lumen of the would-be suprachiasmatic region of the third ventricle. In 70% of the animals examined histologically during the latter phase of development and as adults, regulation occurred and the brains were largely comparable with those of controls. However, in the remaining mutant mice, the overall size of either, or sometimse both, SCN was much reduced. The basal (but not the apical) dendrites of SCN neurons failed to develop fully. Some basal dendrites normally invade the optic chiasm below. In several mutant animals one or the other SCN had greatly increased numbers of cells, while the contralateral one had diminished numbers. These observations suggest that regular formation of the suprachiasmatic region of the hypothalamus and especially the suprachiasmatic nuclei, may depend during development upon the presence of the eye or the subjacent optic axons.

Animals↗

PubMed1978

Histochemical, pharmacological and microspectrofluorometric analysis of new sites of serotonin localization in the rat hypothalamus.

Histopharmacological and microspectrofluorometric analysis was applied to the rat hypothalamus for the identification of serotonin. Neuronal perikarya, especially certain of those of the endocrinologically important arcuate nucleus, were seen to contain a yellow histofluorescence which possessed spectral characteristics consistent with the serotonin fluorophor. Serotonin was localized also within varicosities of various hypothalamic nuclei, as well as within an ependymally associated network of fibers. Pharmacological administration of the serotonin precursor, tryptophan, resulted in the identification of serotonin histofluorescence and spectra within hypothalamic neurons yielding evidence that these neurons possess the capacity to synthesize serotonin. These newly described cells may be of importance to the regulatin of hypothalamically mediated endocrine function due to their neuroanatomical placement in the medical basal hypothalamus.

Animals↗

PubMed1981

Immunohistochemical analysis of magnocellular elements in rat hypothalamus: distribution and numbers of cells containing neurophysin, oxytocin, and vasopressin.

A cell-by-cell analysis of the magnocellular elements in hypothalami of fifty Long-Evans (normal) and Brattleboro (diabetes insipidus) rats was done using the unlabeled antibody enzyme technique (PAP) with primary antisera directed against oxytocin (OXY), vasopressin (ADH), and the neurophysins. The magnocellular neurons of the hypothalamus were found in the supraoptic (SON), paraventricular (PVN), and anterior commissural (ACN) nuclei, a number of accessory nuclei, and as individual cells in the anterior hypothalamic area. SON was divided by the optic tract into the principal part and retrochiasmatic SON. In retrochiasmatic SON a majority of the cells contained vasopressin. Within the principal part of SON oxytocin-producing cells tended to be found rostrally and dorsally, while the vasopressin cells were more common caudally and ventrally. PVN was divided into three subnuclei, the medial, lateral, and posterior subnuclei, on the basis of cellular morphology and peptide content. The magnocellular cells of the medial and lateral PVN were closely packed together and nearly round, while those of posterior PVN were more separated and fusiform in shape with their long axis running in a medio-lateral direction. Medial PVN consisted primarily of oxytocin-producing cells, while lateral PVN was formed by a core of vasopressin-producing cells with a rim of oxytocin cells. Posterior PVN contained largely oxytocin-producing cells. Both ADH and OXY cells were found in the accessory nuclei. In the Long-Evans rat the SON had, on the average, 1443 OXY and 3236 ADH cells; the PVN had 1174 OXY and 976 ADH cells; and the accessory magnocellular groups in the hypothalamus (including the ACN) had 1286 OXY and 552 ADH cells. The Brattleboro strain animal had similar numbers of cells in these nuclei. (The cells which contain ADH in normal animals were identified in the Brattleboro rat as large, neurophysin-negative cells.) Thus, a large fraction of the magnocellular oxytocin- and vasopressin-producing cells in the rat were located outside of the PVN and SON. One accessory cell group in particular, ACN, had 616 OXY cells, or about 50% as many as PVN. In each nucleus the sum of the numbers of OXY and ADH cells was approximately the number of neurophysin cells.

Animals↗

PubMed1984

Immunoreactive leu-enkephalin in the monkey hypothalamus including observations on its ultrastructural localization in the paraventricular nucleus.

The distribution of immunoreactive leu-enkephalin neurons and fibers in the monkey hypothalamus, including ultrastructural localization in the paraventricular nucleus (PVN), was examined with the peroxidase-antiperoxidase immunocytochemical method. Immunoreactive leu-enkephalin cell bodies and fibers were present in the PVN, the region of the dorsal nucleus and nucleus of the anterior commissure, the dorsomedial nucleus, ventromedial nucleus, and lateral hypothalamus. Within the PVN labeled cells were found mostly in the medial parvocellular region, and a smaller proportion including some large cells was present in the lateral, and dorsolateral zones. Immunoreactive neurons contained numerous large granular vesicles (LGV) which ranged from 63 to 235 nm in size, suggesting that at least some enkephalin-containing neurons belong to the population of neurosecretory cells. Positive neurons were postsynaptic to four types of unlabeled axon terminals. Leu-enkephalin-containing fibers (some of which were myelinated) and boutons contained small clear vesicles and numerous LGV. Axon terminals made synaptic contacts with the cell bodies, primary and distal dendrites of unlabeled neurons. The findings show that enkephalin-containing neurons in the PVN integrate a variety of neuronal inputs and provide morphological evidence for the inhibiting influence of enkephalins on the firing rate of PVN neurons. It may be speculated that the effects of opioids on the release of vasopressin and other substances possibly originating from PVN neurons may be regulated in part within the nucleus by locally synapsing axons belonging to enkephalin-containing neurons.

Animals↗

PubMed1988

Somatostatin-containing neuron systems in the rat hypothalamus: retrograde tracing and immunohistochemical studies.

By employing a combination of the immunohistochemistry for somatostatin (SRIF) and retrograde tracing with biotinylated wheat germ agglutinin (b-WGA) injected into the posterior pituitary (group 1) or into the median eminence (group 2), functional topography of hypothalamic SRIF neurons was determined in the rat hypothalamus. In group 1, large numbers of WGA-labeled neurons appeared in the rostral periventricular region and in the magnocellular division of the paraventricular and supraoptic nuclei; none of them were SRIF immunoreactive. In group 2, WGA-labeled neurons were numerous in the rostral periventricular region, the parvicellular division of the paraventricular nucleus, and the arcuate nucleus; most of the WGA-labeled neurons in the rostral periventricular region and some in the paraventricular nucleus were SRIF immunoreactive, but none in the arcuate nucleus showed immunoreactivity for SRIF. It is concluded that, in the rat hypothalamus, the locations of neurons containing hypophysiotrophic SRIF are confined within the rostral periventricular region and the parvicellular paraventricular nucleus. Our results do not support previous suggestions that SRIF immunoreactive axons innervate the posterior lobe of the pituitary.

Animals↗

PubMed1989

Distribution and morphometric characteristics of oxytocin- and vasopressin-immunoreactive neurons in the rabbit hypothalamus.

The distribution, morphological features, and morphometric characteristics of cell bodies producing oxytocin (OT) and vasopressin (AVP) were studied in the rabbit hypothalamus by means of a conventional immunoperoxidase method. The aim of the present study was to determine the existence or not of a species-specific OT-cell group that might be involved in the dense OT innervation of the intermediate lobe in the leporidae. No OT-cell group clearly distinct from the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei was found, even in colchicine-treated animals. Most immunoreactive perikarya were found within these nuclei. In addition, small AVP neurons occurred in the suprachiasmatic nucleus. In the SON, the predominant, tightly packed AVP cells occupied the ventral part of the nucleus, whereas OT neurons were dorsolaterally located. The PVN presented a loose organization without any obvious subdivision. OT cells, which predominated, occupied the medial part of the nucleus. The PVN had a prominent rostral anterobasal extension composed mainly of OT cells. Laterally to the nucleus, numerous large AVP neurons, with few and smaller OT cells, dispersed along the neurosecretory tract without forming definite cell clusters. AVP cell bodies had a rough granular aspect contrasting with the smooth and fine one of OT cells. Spinelike processes were rarely observed on the perikarya, except on large scattered AVP neurons, but frequently covered the proximal dendrites of both types of neurons. Throughout the hypothalamus, OT neurons had definitely smaller mean somal areas and were more homogeneous in size than AVP cells.

Animals↗

PubMed1990

Projections from the lateral geniculate nucleus to the hypothalamus of the Mongolian gerbil (Meriones unguiculatus): an anterograde and retrograde tracing study.

The lateral geniculate nucleus of the thalamus sends efferents to the hypothalamic suprachiasmatic nucleus, which is involved in generation and entrainment of several circadian rhythms. It seems reasonable to believe that the lateral geniculate conveys visual information about the length of the photoperiod to the circadian oscillator. In order to study in more detail the topographical relationship between the lateral geniculate and the suprachiasmatic nucleus, anterograde tracing with Phaseolus vulgaris leucoagglutinin (PHA-L) and retrograde tracing with wheatgerm agglutinin coupled to horseradish peroxidase (WGA-HRP) were performed in the gerbil. After iontophoretic injections of PHA-L in the lateral geniculate, a large number of PHA-L-immunoreactive fibers and nerve terminals were observed in the ventrolateral part of the suprachiasmatic nucleus. Nerve fibers were also present in the ventromedial and dorsolateral portions, particularly in the caudal half of the nucleus. PHA-L-immunoreactive nerve fibers continued outside the borders of the suprachiasmatic nucleus to the adjacent anterior hypothalamic, the periventricular, and the subparaventricular areas. A moderate number of fibers entered the lateral hypothalamic area and the tuber cinerum via the optic tract and chiasm. Moreover, the paraventricular nucleus, the supraoptic nucleus, the medial preoptic area, the lateral preoptic area, and the supramammillary nucleus contained a few labeled fibers. In all parts of the hypothalamus receiving an input from the lateral geniculate, fine beaded immunoreactive fibers with varicosities and nerve terminals were observed, some of which were found in close apposition to hypothalamic neurons. Only after labeling of neurons in the intergeniculate leaflet of the lateral geniculate nucleus, fibers were found in the hypothalamus. This topographical organization of the geniculohypothalamic pathway was supported by retrograde tracing after injections of WGA-HRP in the suprachiasmatic area. In these experiments, retrograde labeled neurons were observed in the intergeniculate leaflet and, in agreement with the anterograde studies, most of labeling was observed in the ipsilateral side. These results confirm that the suprachiasmatic nucleus receives a substantial input from the intergeniculate leaflet of the lateral geniculate. Moreover, the present data demonstrate that the suprachiasmatic nucleus is not the only nucleus that receives a direct visual input. Thus other hypothalamic areas might be influenced by a direct rhythmic neuronal input as well.

Animals↗

PubMed1990

A vasopressin and oxytocin containing nucleus in the pig hypothalamus that shows neuronal changes during puberty.

A vasopressin and oxytocin containing nucleus is described for the first time in the pig hypothalamus. It is located near the third ventricle, just dorsal to the suprachiasmatic nucleus, and consists of magnocellular neurons, similar to those of the supraoptic nucleus and paraventricular nucleus. Morphometric analysis of neuronal number, size, density, and volume was performed at four different ages: 1 day, 7 weeks, 16 weeks, and 30 weeks postnatally. No sex difference in these parameters was observed. In this period the volume of the nucleus increased gradually from 6.6 x 10(-3) to 54.2 x 10(-3) mm3. One day after birth 1,215 +/- 191 (mean +/- SEM) neurons were present in the vasopressin and oxytocin containing nucleus, followed by a decrease to 771 +/- 80 neurons at 7 weeks and 697 +/- 116 at 16 weeks. Between 16 and 30 weeks (puberty) there was a dramatic increase in neuron number up to 1,765 +/- 214 neurons. This increase in the number of vasopressin and oxytocin containing neurons in the pig hypothalamus is much later in development than has ever been reported so far.

Aging↗

PubMed1992

Distribution of vasopressin and oxytocin cells and fibres in the hypothalamus of the domestic pig (Sus scrofa).

The distribution of vasopressin and oxytocin cells and fibres was studied in the pig hypothalamus by means of monoclonal antibodies, in order to prevent the non-specific staining that is characteristic for the pig. The nucleus circularis is described for the first time in the pig hypothalamus and consists of both oxytocin and vasopressin containing cell bodies. A cell group, characteristic for the pig and horse, lateral and caudal from the vascular organ of the lamina terminalis, was found to be continuous with the supraoptic nucleus. It contained only oxytocin and it is proposed that this group be called pars dorsomedialis of the supraoptic nucleus. In this study, vasopressin has been demonstrated for the first time in the suprachiasmatic nucleus of the pig. The discussion focusses on how our findings in the pig differ from vasopressin and oxytocin systems identified in other mammalian species.

Animals↗

PubMed1994

Topography of neurons expressing luteinizing hormone-releasing hormone gene transcripts in the human hypothalamus and basal forebrain.

The distribution of neurons expressing luteinizing hormone-releasing hormone (LHRH) gene transcripts was mapped in the human hypothalamus and basal forebrain by in situ hybridization and computer-assisted microscopy. Hypothalamic blocks were dissected from five adult males and one adult female and snap frozen in isopentane. The blocks were serially sectioned either in the coronal or in the sagittal plane at a thickness of 20 microns. Approximately every twentieth section was incubated with a 35S-labeled cDNA probe complementary to LHRH mRNA. Specificity was confirmed by hybridization of adjacent sections with a probe targeted to the gonadotropin-associated protein (GAP) region of LHRH messenger ribonucleic acids (mRNA). Maps of neurons containing LHRH mRNA were manually digitized with the aid of an image-combining computer microscope system. We report a much wider distribution and greater numbers of LHRH neurons than have been previously described in the human brain. Three morphological subtypes were observed based on cell size and labeling density: 1) small, heavily labeled, oval or fusiform neurons, located primarily in the medial basal hypothalamus, ventral preoptic area, and periventricular zone; 2) small, oval, sparsely labeled neurons located in the septum and dorsal preoptic region and scattered from the bed nucleus of the stria terminalis to the amygdala ("extended amygdala"); and 3) large round neurons (> 500 microns 2 sectional profile area), intermediate in labeling density, scattered within the magnocellular basal forebrain complex, extended amygdala, ventral pallidum, and putamen. The pronounced differences in morphology, labeling density, and location of the three subtypes suggest that distinct functional subgroups of LHRH neurons exist in the human brain.

Adolescent↗

PubMed2005

Regionalization of the anterior hypothalamus in the chick embryo.

Loss-of-function experiments in mice have shown that the transcription factors Sim1, Otp, Sim2, and Brn2 form a cascade essential for the differentiation of neuroendocrine cells of the anterior hypothalamus that produce vasopressin, oxytocin, somatostatin (SS), thyrotropin-releasing hormone (TRH), and corticotropin-releasing hormone (CRH). Very little is known about how the differentiation of these cell types is regulated in chick. Here, we report the cloning of the chick homolog of Otp. Moreover, we have systematically compared the expression of Sim1, Sim2, Brn2, and Otp with that of the markers of terminal differentiation TRH, SS, CRH, vasotocin, and mesotocin during development of chick embryos. We have found that the cell types studied generally develop in domains expressing these transcriptional regulators but that the pattern of neuronal differentiation and the spatial distribution of some regulators were not the same as in mice. Our results provide a framework useful for the functional analysis of hypothalamus development in chick.

Animals↗

PubMed1995

Immunohistochemical localization of glucose transporters (GLUT1 and GLUT3) in the rat hypothalamus.

Immunohistochemical localization of the glucose transporters was studied in the rat hypothalamus by using a specific rabbit antiserum raised against either the isoform 1 (GLUT1) or the isoform 3 (GLUT3). Immunoreactive staining for GLUT1 was found in glia cells and capillaries, whereas positive staining for GLUT3 occurred mainly in neurons and partly in ependymal cells. Double immunostaining indicated that a small population of GLUT1-positive cells were reactive for glial fibrillary acidic protein, a marker for astrocytes. Another doubly stained section showed that GLUT1-positive glia cells were never stained with an OX42 antibody, a marker for microglia cells. Neurons staining positively for GLUT3, often large in cell size, were confined mainly in the lateral hypothalamic area and partly in the dorsomedial and periventricular hypothalamic nuclei. Possible significance of these two glucose transporters in the hypothalamus is briefly discussed.

Amino Acid Sequence↗

PubMed1980

Effects of 2-deoxy-D-glucose on multi-unit activity of the hypothalamus in rats.

Multi-unit activity was recorded from the diencephalon of unanesthetized, immobilized female rats. Less than 50% of the recording sites in the hypothalamus showed changes in the level of multi-unit activity to changes in the level of glucose utilization induced by injection of 2-deoxy-D-glucose (2-DG). It was also not possible to specificy an area within the hypothalamus in which there was a concentration of recording sites which consistently responded to 2-DG. Relatively large amplitude, long duration changes in multi-unit activity levels were also observed in the optic tract, suggesting that 2-DG induced a widespread cellular metabolic response to glucoprivation which may not have been related to the regulation of feeding.

Animals↗

PubMed1980

Effects of morphine on sensory-evoked responses recorded from central gray, reticular formation, thalamus, hypothalamus, limbic system, basal ganglia, dorsal raphe, locus ceruleus, and pineal body.

Field potential recordings of acoustic and photic-evoked responses were obtained from 15 brain sites of freely behaving unanesthetized rats previously implanted stereotaxically with permanent electrodes. Several dosages of morphine (1, 5, 10, 30, and 50 mg/kg) were examined. The activities recorded from all the structures in this study, except the cochlear nucleus (CoN), were affected by morphine. Different sensitivities to morphine threshold were observed between structures, and several structures exhibited dose-related patterns (ventromedial hypothalamus (VMH), caudate nuucleus (CN), central gray (CG), hippocampus (Hipp), and lateral septum (Spt)). Several brain sites, after the initial dose of morphine, did not recruit more responses to subsequent doses of the drug, ie, exhibited all-or-none responses (pineal body (PB), medial thalamus (MTh), anterior hypothalamus (AH), mesencephalic reticular formation (MRF), and the dorsal raphe (DR)). In some structures, morphine induced increases in the response amplitudes, while in other sites decreases in response amplitudes were elicited. Biphasic responses, ie, increases in response amplitude after low doses of the drug and decreases in response amplitude after higher dosages, were also observed (VMH, CN, DR, CG, and MRF). The acoustic-evoked responses were affected by morphine more than the photic responses. The present observations indicated that 1) morphine exerts effects in many parts of the central nervous system (CNS); 2) some structures are more sensitive to morphine than others; 3) only a few structures exhibit dose-related patterns and, thus, may represent sites of direct morphine action; 4) some structures exhibit all-or-none responses; and 5) morphine depressed activity in some structures and increased activity in others, ie, morphine elicited different effects in different structures.

Acoustic Stimulation↗

PubMed1991

Tamoxifen fails to block estradiol accumulation, yet is weakly accumulated by the juvenile zebra finch anterior hypothalamus: an autoradiographic study.

In experiment 1, we used autoradiographic procedures to examine whether tamoxifen could displace 3H-estradiol labeling in the anterior hypothalamus and the caudal nucleus of the ventral hyperstriatum (HVc) of ovariectomized 20-day-old female zebra finches. There was no significant reduction in labeling of cells by 3H-estradiol in birds preinjected with unlabeled tamoxifen. In experiment 2, we found that injections of 3H-tamoxifen caused-weak labeling of cells in the anterior hypothalamus of 20-day-old male and female zebra finches. These results are compatible with the idea that tamoxifen does not block the action of estradiol in the brain of zebra finches, and suggest that the effects of early tamoxifen treatment on the morphology of the song system may reflect central actions of tamoxifen.

Animals↗

PubMed1996

Developmental changes and molecular weight of immunoreactive glucocorticoid receptor protein in the ovine fetal hypothalamus and pituitary.

Parturition in the sheep is initiated by increased activity of the hypothalamus-pituitary-adrenal (HPA) axis in the fetus. Late in gestation, ACTH secretion increases because of increased stimulation to the HPA axis and a decreased cortisol negative feedback inhibition of ACTH secretion. We hypothesized that this alteration in cortisol negative feedback efficacy might result from a decrease in glucocorticoid receptor (GR) protein in the hypothalamus or pituitary. Fetal hypothalami and pituitaries were collected from healthy animals (74 days gestation to 4 weeks postnatally) and analyzed by Western blot for immunoreactive GR (iGR) protein. We detected two forms of iGR: a form consistent in molecular weight with the GR, and a form which is approximately half the size of the full-length receptor. We were not able to detect a developmental change in the intensity of immuno-staining for the full-length receptor protein in these tissues. However, the proportion of half-length relative to full-length GR increased significantly with developmental age. Exogenous estrogen, androgen, or tamoxifen treatment did not significantly affect the expression of GR protein. In conclusion, we detected a 97 kD iGR protein corresponding to the full-length receptor and a 45 kD iGR protein, or half-length receptor. The proportion of this half-length receptor protein relative to the full-length receptor increased significantly with development.

Amino Acid Sequence↗

PubMed1996

Dispersion patterns of clonally related cells during development of the hypothalamus.

To identify patterns of cell generation and migration during early development of the hypothalamus, clones were marked with a library of retroviral vectors. The majority of clones analyzed at Embryonic Days 8-10 were simple radial columns. A small number of clones had one of three unusual patterns of dispersion. One unusual clone type was bilaterally symmetric across the ventral forebrain. Another was composed of four or more columns of cells arranged in a line along the ventricular surface, and a third was composed of cells that appeared to disperse from the typical radial columns in a plane parallel to the ventricular surface. The appearance of the rare clones suggests that there are different mechanisms of clonal dispersion during early development of the ventral forebrain that could play a role in patterning the hypothalamus.

Animals↗