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PubMed2001

Orbitomedial prefrontal cortical projections to hypothalamus in the rat.

A previous study in the rat revealed that distinct orbital and medial prefrontal cortical (OMPFC) areas projected to specific columns of the midbrain periaqueductal gray region (PAG). This study used anterograde tracing techniques to define projections to the hypothalamus arising from the same OMPFC regions. In addition, injections of anterograde and retrograde tracers were made into different PAG columns to examine connections between hypothalamic regions and PAG columns projected upon by the same OMPFC regions. The most extensive patterns of hypothalamic termination were seen after injection of anterograde tracer in prelimbic and infralimbic (PL/IL) and the ventral and medial orbital (VO/MO) cortices. Projections from rostral PL/IL and VO/MO targeted the rostrocaudal extent of the lateral hypothalamus, as well as lateral perifornical, and dorsal and posterior hypothalamic areas. Projections arising from caudal PL/IL terminated within the dorsal hypothalamus, including the dorsomedial nucleus and dorsal and posterior hypothalamic areas. There were also projections to medial perifornical and lateral hypothalamic areas. In contrast, it was found that anterior cingulate (AC), dorsolateral orbital (DLO), and agranular insular (AId) cortices projected to distinct and restricted hypothalamic regions. Projections arising from AC terminated within dorsal and posterior hypothalamic areas, whereas DLO and AId projected to the lateral hypothalamus. The same OMPFC regions also projected indirectly, by means of specific PAG columns, to many of the same hypothalamic fields. In the context of our previous findings, these data indicate that, in both rat and macaque, parallel but distinct circuits interconnect OMPFC areas with specific hypothalamic regions, as well as PAG columns.

Animals↗
PubMed2006

Innervation of the paraventricular nucleus of the thalamus from cocaine- and amphetamine-regulated transcript (CART) containing neurons of the hypothalamus.

The paraventricular nucleus of the thalamus (PVT) is a midline thalamic nucleus with heavy projections to the nucleus accumbens and other limbic regions. Previous studies have shown that the PVT contains fibers immunoreactive for cocaine- and amphetamine-related transcript (CART). The purpose of the present study was to determine the location of CART neurons innervating the PVT of the rat by using retrograde tracing with cholera toxin B (CTb) combined with immunofluorescence for CTb and CART (amino acid sequence 55-102). Immunohistochemical analysis of CART in the dorsal thalamus showed that the PVT is densely innervated by CART fibers whereas adjacent midline and intralaminar thalamic nuclei are unlabeled. Injections of CTb in the dorsal midline thalamus retrogradely labeled neurons in several areas of the hypothalamus and brainstem which also contained CART neurons. The largest number of double-labeled neurons (CTb/CART) was found in the arcuate nucleus of the hypothalamus. CTb/CART neurons were also found in the lateral hypothalamus, zona incerta, and periventricular hypothalamus. These results indicate that the arcuate nucleus is a major source of CART fibers in the PVT. CART neurons in the arcuate nucleus monitor circulating hormonal signals and may regulate food intake and hypothalamic-pituitary-adrenal (HPA) activity. Consequently, CART neurons in the arcuate nucleus may transmit signals to the PVT which in turn may influence limbic regions involved in regulating food intake and the HPA.

Animals↗
PubMed2006

Distribution of MT1 melatonin receptor immunoreactivity in the human hypothalamus and pituitary gland: colocalization of MT1 with vasopressin, oxytocin, and corticotropin-releasing hormone.

Melatonin is implicated in numerous physiological processes, including circadian rhythms, stress, and reproduction, many of which are mediated by the hypothalamus and pituitary. The physiological actions of melatonin are mainly mediated by melatonin receptors. We here describe the distribution of the melatonin receptor MT1 in the human hypothalamus and pituitary by immunocytochemistry. MT1 immunoreactivity showed a widespread pattern in the hypothalamus. In addition to the area of the suprachiasmatic nucleus (SCN), a number of novel sites, including the paraventricular nucleus (PVN), periventricular nucleus, supraoptic nucleus (SON), sexually dimorphic nucleus, the diagonal band of Broca, the nucleus basalis of Meynert, infundibular nucleus, ventromedial and dorsomedial nucleus, tuberomamillary nucleus, mamillary body, and paraventricular thalamic nucleus were observed to have neuronal MT1 receptor expression. No staining was observed in the nucleus tuberalis lateralis and bed nucleus of the stria terminalis. The MT1 receptor was colocalized with some vasopressin (AVP) neurons in the SCN, colocalized with some parvocellular and magnocellular AVP and oxytocine (OXT) neurons in the PVN and SON, and colocalized with some parvocellular corticotropin-releasing hormone (CRH) neurons in the PVN. In the pituitary, strong MT1 expression was observed in the pars tuberalis, while a weak staining was found in the posterior and anterior pituitary. These findings provide a neurobiological basis for the participation of melatonin in the regulation of various hypothalamic and pituitary functions. The colocalization of MT1 and CRH suggests that melatonin might directly modulate the hypothalamus-pituitary-adrenal axis in the PVN, which may have implications for stress conditions such as depression.

Adult↗
PubMed1978

Development of the diencephalon in the rat. I. Autoradiographic study of the time of origin and settling patterns of neurons of the hypothalamus.

Groups of pregnant rats were injected with two successive daily doses of 3H-thymidine from gestational day 13 (E13 + 14) until the day before birth (E21 + 22). This double labelling procedure was combined with an injection schedule of a single day delay between groups (E13 + 14; E14 + 15; E15 + 16 . .). The two injections assured the comprehensive labelling of practically all neurons of a given structure prior to the onset of their differentiation (comprehensive labelling), whereas the progressive daily delay in injections made it possible to estimate the proportion of neurons formed in various regions of the hypothalamus on a single day. Hypothalamic areas or nuclei were assigned into four classes on the basis of their cytogenetic isochronicity. Structures composed of the earliest arising (class 1) neurons constitute a lateral tier that includes the lateral preoptic and lateral hypothalamic areas, and the lateral mammillary nucleus. Structures composed of early arising (class 2) neurons form a heterogeneous collection of nuclear systems, including the paraventricular, internuclear and supraoptic magnocellular neurons, and several intermediate tier nuclei of the anterior and posterior hypothalamus. The late arising (class 3) and latest arising (class 4) nuclei constitute a periventricular system anteriorly and a more extensive region posteriorly. The latter two nuclear systems may constitute the hypophysiotropic area of the hypothalamus. The nuclei of the mammillary system, which are produced sequentially, are distinguished from other hypothalamic structures by their more rapid generation time. Internuclear labelling gradients were used to infer the neuroepithelial site of origin and settling pattern of neurons. Common sites of origin were indicated for the following structures: the magnocellular neurohypophysial neurons; the neurons of the dorsomedial and ventromedial nuclei; and the neurons of the tubermammillary and arcuate nuclei. The sites of origin of these groups of nuclei were related to specialized ventricular linings in the mature hypothalamus.

Animals↗
PubMed1987

Mapping of neuropeptide Y-like immunoreactivity in the feline hypothalamus and hypophysis.

The distribution of neuropeptide Y (NPY) in the cat hypothalamus and hypophysis was studied with the indirect immunofluorescence technique of Coons and co-workers (Coons, Leduc, and Connolly: J. Exp. Med. 102:49-60, 1955), which provided a detailed map of NPY-like immunoreactive neurons. The immunolabelling was detected in cell bodies, fibers, and terminallike structures widely distributed throughout the whole hypothalamus. A large population of medium-sized NPY-like immunoreactive cell bodies was localized in the area of arcuate nucleus. The number of immunoreactive cell bodies visualized was dramatically increased after intracerebroventricular injections of colchicine. Numerous immunolabelled cell bodies were also visible in the median eminence and scattered in the lateral hypothalamic area. Dense plexuses of NPY-immunoreactive fibers were observed in the arcuate nucleus, internal layer of median eminence, periventricular zone, and paraventricular nucleus. Other regions of hypothalamus displaying numerous NPY-like immunoreactive fibers included dorsal and ventrolateral hypothalamic areas. In contrast, certain hypothalamic areas were almost devoid of NPY-like immunoreactive fibers-namely, the mammillary bodies and suprachiasmatic nucleus. Finally, in neurohypophysis, bright immunofluorescent fibers were observed along the pituitary stalk and penetrating the neural lobe. These results suggest the widespread distribution of the NPY-containing neuronal systems in the cat hypothalamus and hypophysis.

Animals↗
PubMed1987

Distribution of neuropeptide Y-like immunoreactivity in the hypothalamus of the adult golden hamster.

The distribution of neuropeptide Y (NPY)-like immunoreactivity within the hypothalamus of the adult golden hamster was investigated with conventional immunohistochemical techniques. Neuropeptide Y immunoreactive cell bodies were found in greatest numbers in the arcuate nucleus while a few stained perikarya were seen in the internal and subependymal zones of the median eminence. Isolated perikarya were observed in the anterior commissure and supracommissural portion of the interstitial nucleus of the stria terminalis. Immunoreactive axons were located throughout the hypothalamus with the highest concentrations in the subependymal and internal zones of the median eminence, the interstitial nucleus of the stria terminalis, the medial preoptic area, and in the following nuclei: periventricular, suprachiasmatic, paraventricular, perifornical, median preoptic, and arcuate. Moderate to dense plexuses of immunoreactive fibers were observed in the anterior, lateral, and posterior hypothalamic areas and in the infundibular stalk. The supraoptic nucleus and lateral preoptic area displayed a small number of labeled axons whereas the ventromedial nucleus contained only a few fibers. NPY immunoreactive fibers were present in the optic tract and in the dorsomedial aspect of the optic chiasm. Labeled fibers penetrated the ependymal lining of the third ventricle throughout the ventral aspect of the periventricular zone. Additional fibers were observed in the pia lining the ventral aspect of the hypothalamus. This systematic analysis of hypothalamic NPY immunoreactivity in the adult golden hamster suggests that a portion of the labeled fibers display a distribution that is similar to previously described noradrenergic fibers in the hypothalamus.

Animals↗
PubMed1987

Some anatomical observations on the projections from the hypothalamus to brainstem and spinal cord: an HRP and autoradiographic tracing study in the cat.

The hypothalamus is closely involved in a wide variety of behavioral, autonomic, visceral, and endocrine functions. To find out which descending pathways are involved in these functions, we investigated them by horseradish peroxidase (HRP) and autoradiographic tracing techniques. HRP injections at various levels of the spinal cord resulted in a nearly uniform distribution of HRP-labeled neurons in most areas of the hypothalamus except for the anterior part. After HRP injections in the raphe magnus (NRM) and adjoining tegmentum the distribution of labeled neurons was again uniform, but many were found in the anterior hypothalamus as well. Injections of 3H-leucine in the hypothalamus demonstrated that: The anterior hypothalamic area sent many fibers through the medial forebrain bundle (MFB) to terminate in the ventral tegmental area of Tsai (VTA), the rostral raphe nuclei, the nucleus Edinger-Westphal, the dorsal part of the substantia nigra, the periaqueductal gray (PAG), and the interpeduncular nuclei. Further caudally a lateral fiber stream (mainly derived from the lateral parts of the anterior hypothalamic area) distributed fibers to the parabrachial nuclei, nucleus subcoeruleus, locus coeruleus, the micturition-coordinating region, the caudal brainstem lateral tegmentum, and the solitary and dorsal vagal nucleus. Furthermore, a medial fiber stream (mainly derived from the medial parts of the anterior hypothalamic area) distributed fibers to the superior central and dorsal raphe nucleus and to the NRM, nucleus raphe pallidus (NRP), and adjoining tegmentum. The medial and posterior hypothalamic area including the paraventricular hypothalamic nucleus (PVN) sent fibers to approximately the same mesencephalic structures as the anterior hypothalamic area. Further caudally two different fiber bundles were observed. A medial stream distributed labeled fibers to the NRM, rostral NRP, the upper thoracic intermediolateral cell group, and spinal lamina X. A second and well-defined fiber stream, probably derived from the PVN, distributed many fibers to specific parts of the lateral tegmental field, to the solitary and dorsal vagal nuclei, and, in the spinal cord, to lamina I and X, to the thoracolumbar and sacral intermediolateral cell column, and to the nucleus of Onuf. The lateral hypothalamic area sent many labeled fibers to the lateral part of the brainstem and many terminated in the caudal brainstem lateral tegmentum, including the parabrachial nuclei, locus coeruleus, nucleus subcoeruleus, and the solitary and dorsal vagal nuclei.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Type B monoamine-oxidase-containing cells and fibers in the cat hypothalamus demonstrated by an improved enzyme histochemical method.

The present study, using a diaminobenzidine (DAB)-coupled peroxidation method, examined the distribution and morphological characteristics of neuronal structures containing type B monoamine oxidase (MAO-B) in the cat hypothalamus. Large and intensely stained, distinctive MAO-B-positive cells, multipolar and with long dendritic arbors, were principally distributed in the ventral hypothalamus extending from A7 to A12.5 of the Horsley-Clarke plane. These cells were located caudally in the ventral surface of the brain including the tuberomamillary nucleus (TM) and the region surrounding the mamillary nuclei. Rostrally, they were aggregated in the area surrounding the fornix, particularly in the lateral perifornical region, and dispersed in the anterior mamillary nucleus, lateral hypothalamic area (HLA), and the medial tip of the entopeduncular nucleus. The most rostral positive cell group was identified in a narrow space between the optic tract and the entopeduncular nucleus at the A12.5 level. In addition to these large cells, the present study disclosed the presence of "small" to "very small" MAO-B-positive cells in the area surrounding the mamillary recess and the lateral part of the caudal arcuate nucleus. Distinct MAO-B-stained fibers were identified in all regions of the hypothalamus. A large number of thick labeled fibers were observed in the ventral hypothalamus including the TM and premamillary nucleus and posterior and lateral hypothalamic areas. A dense network of MAO-B-positive terminal-like fibers was observed in the dorsomedial nucleus where very small labeled cells were scattered. Many intensely stained thick and straight fibers were seen running ventrolaterally in the anterior part of the HLA and in the narrow space between the entopeduncular nucleus and optic tract. In the area of the tuber cinereum and the ventral part of the HLA, there were many positive fibers cut transversely, possibly projecting to the more anterior parts of the brain such as the diagonal band of Broca or septal nuclei.

Animals↗
PubMed1993

Noradrenergic innervation of the hypothalamus of rhesus monkeys: distribution of dopamine-beta-hydroxylase immunoreactive fibers and quantitative analysis of varicosities in the paraventricular nucleus.

The distribution of noradrenergic processes within the hypothalamus of rhesus monkeys (Macaca mulatta) was examined by immunohistochemistry with an antibody against dopamine-beta-hydroxylase. The results revealed that the pattern of dopamine-beta-hydroxylase immunoreactivity varied systematically throughout the rhesus monkey hypothalamus. Extremely high densities of dopamine-beta-hydroxylase-immunoreactive processes were observed in the paraventricular and supraoptic nuclei, while relatively lower levels were found in the arcuate and dorsomedial nuclei and in the medial preoptic, perifornical, and suprachiasmatic areas. Moderate levels of dopamine-beta-hydroxylase immunoreactivity were found throughout the lateral hypothalamic area and in the internal lamina of the median eminence. Very few immunoreactive processes were found in the ventromedial nucleus or in the mammillary complex. Other midline diencephalic structures were found to have high densities of dopamine-beta-hydroxylase immunoreactivity, including the paraventricular nucleus of the thalamus and a discrete subregion of nucleus reuniens, the magnocellular subfascicular nucleus. A moderate density of dopamine-beta-hydroxylase immunoreactive processes were found in the rhomboid nucleus and zona incerta whereas little dopamine-beta-hydroxylase immunoreactivity was found in the fields of Forel, nucleus reuniens, or subthalamic nucleus. The differential distribution of dopamine-beta-hydroxylase-immunoreactive processes may reflect a potential role of norepinephrine as a regulator of a variety of functions associated with the nuclei that are most heavily innervated, e.g., neuroendocrine release from the paraventricular and supraoptic nuclei, and gonadotropin release from the medial preoptic area and mediobasal hypothalamus. Additionally, quantitative analysis of dopamine-beta-hydroxylase-immunoreactive varicosities was performed on a laser scanning microscope in both magnocellular and parvicellular regions of the paraventricular nucleus of the hypothalamus. The methodology employed in this study allowed for the high resolution of immunoreactive profiles through the volume of tissue being analyzed, and was more accurate than conventional light microscopy in terms of varicosity quantification. Quantitatively, a significant difference in the density of dopamine-beta-hydroxylase-immunoreactive varicosities was found between magnocellular and parvicellular regions, suggesting that parvicellular neurons received a denser noradrenergic input. These differential patterns may reflect an important functional role for norepinephrine in the regulation of anterior pituitary secretion through the hypothalamic-pituitary-adrenal stress axis.

Animals↗
PubMed1993

Developmental study of GnRH neuronal projections to the medial basal hypothalamus of the male Djungarian hamster.

The present study in the male Djungarian hamster determined the neuroanatomical distribution and morphology of gonadotropin-releasing hormone (GnRH) neurons which innervate the medial basal hypothalamus during sexual maturation. Prepubertal, peripubertal, and postpubertal males were perfused, brains were removed, and crystals of the fluorescent tract tracer, DiI, were implanted directly into the median eminence of the brain. Eight weeks later, brains were sectioned and processed for GnRH immunofluorescence. At all ages, GnRH cell bodies were bipolar or unipolar; both subtypes were labeled with DiI in proportion to their respective numbers in each brain region. GnRH perikarya were distributed in a diffuse ventromedial continuum from the septum through the anterior hypothalamus. In prepubertal males, DiI was present in the majority of GnRH neurons (54% of total) that were located in brain regions rostral to and including the medial preoptic area. In lateral and caudal brain areas, fewer GnRH perikarya contained DiI (28% of total or less). With sexual maturation, fewer GnRH somata were labeled with DiI in areas rostral to the hypothalamus. The data suggest that bipolar and unipolar GnRH neurons in the forebrain, rostral to the preoptic area, are major contributors to the GnRH innervation of the median eminence in the male Djungarian hamster. With the onset of puberty, the finding that decreasing numbers of GnRH perikarya directly project to the medial basal hypothalamus suggests that fewer GnRH neurons constitute the final common pathway that controls gonadotropin secretion.

Aging↗
PubMed1993

Efferent projections of the sexually dimorphic area of the gerbil hypothalamus: anterograde identification and retrograde verification in males and females.

Outputs of the sexually dimorphic area (SDA) of the gerbil hypothalamus were identified by injecting Phaseolus vulgaris-leucoagglutinin into the medial or lateral SDA (mSDA, lSDA) in males and females. They were verified by injecting Fluoro-Gold or rhodamine-labeled beads into over half the areas that contained labeled fibers. Both anterograde and retrograde tracing showed that the mSDA and lSDA project to many of the same sites but often to differing degrees. The mSDA projects more heavily than the lSDA to many of their forebrain targets including the ventral part of the lateral septal nucleus, the bed nucleus of the stria terminalis, the medial tuberal area, and the anteroventral periventricular, arcuate, ventromedial and ventral premammillary nuclei of the hypothalamus. The lSDA projects more heavily than the mSDA to many of their mid- and hindbrain targets including the caudal, ventrolateral part of the periaqueductal gray, the retrorubral field, the pedunculopontine tegmental nucleus, and the locus coeruleus. In many other areas of the brain, the projections of the mSDA and lSDA are similar in size. These areas include the substantia innominata, the vascular organ of the lamina terminalis, the anterior amygdala, the posterior hypothalamus, the reuniens and paraventricular nuclei of the thalamus, and the pontine periaqueductal gray lateral to the fourth ventricle. The SDA pars compacta (SDApc), a small cell group embedded in the mSDA of males, projects to many fewer areas than the surrounding mSDA. It was strongly labeled when retrograde tracers were injected into the encapsulated part of the bed nucleus of the stria terminalis, the anteroventral periventricular nucleus, or the mSDA. It was also labeled from the vascular organ of the lamina terminalis, the caudal part of the lateral bed nucleus of the stria terminalis, the lSDA, the area lateral to the mSDA, the arcuate nucleus, the ventral premammillary nucleus, and the ventrolateral part of the ventromedial nucleus of the hypothalamus. Nothing resembling an SDApc was identified during retrograde tracing in females.

Animals↗
PubMed1995

Metabotropic glutamate receptor mGluR5 subcellular distribution and developmental expression in hypothalamus.

The metabotropic glutamate receptor mGluR5 is a G-protein coupled receptor that plays a key role in release of Ca2+ from internal stores via inositol triphosphate mobilization. Western and Northern blot analyses revealed a greatly enhanced expression of mGluR5 in rats during early stages of hypothalamic development compared with the adult. This enhanced developmental expression provides an explanation for the dramatic physiological response of developing neurons to metabotropic glutamate receptor activation and supports the argument that metabotropic glutamate receptors may play an important role in hypothalamic development. During development, expression of the mGluR5 gene was reduced, not only in the hypothalamus but also in other regions of the brain. A differential decrease in mGluR5 protein was found in different brain regions with Western blot analysis. The hypothalamus showed a sixfold decrease in mGluR5 with development, whereas the cortex showed only a threefold decrease. Immunocytochemistry with an affinity-purified antibody against a peptide deduced from the cloned mGluR5 gene revealed selective expression in some regions in the adult hypothalamus. In the adult and developing (postnatal day 10) brain, immunoreactive neurons were found in the suprachiasmatic nucleus, preoptic area, lateral hypothalamus, and mammillary region, areas where the related metabotropic glutamate receptor mGluR1 is also found. In contrast, the ventromedial nucleus, an area critically involved in the regulation of food intake and metabolic balances, showed strong mGluR5 immunoreactivity but no mGluR1 immunoreactivity. Little or no mGluR5 staining was found in the neurosecretory neurons of the paraventricular, supraoptic, and arcuate nuclei. Ultrastructurally, mGluR5 was associated with the cytoplasmic face of the plasmalemma on hypothalamic dendrites, dendritic spines, and neuronal perikarya in the adult. The strongest immunoreactivity was found in patches on the membrane, sometimes associated with the postsynaptic side of synapses and sometimes associated with nonsynaptic dendritic or perikaryal membrane. Intense immunostaining was found on some astrocyte processes surrounding synaptic complexes containing asymmetrical synapses. These astrocytes would be in an ideal position to receive excitatory signals from glutamatergic axons. Unlike the punctate appearance of immunolabeling on neuronal membranes, astrocytes showed continuous staining along the plasma membrane.

Animals↗
PubMed1985

In situ hybridization of putative somatostatin mRNA within hypothalamus of the rat using synthetic oligonucleotide probes.

The distribution of mRNA with high sequence homology to somatostatin mRNA within the periventricular hypothalamus of rat was assessed using in situ hybridization techniques with synthetic oligodeoxyribonucleotide probes, complementary to the 3' coding region of rat somatostatin mRNA. The probes (22- and 24-mers) were 5'-end labeled using T4 polynucleotide kinase and gamma-32P-ATP. They were used either individually or after ligation with T4 DNA ligase to form a 46-mer. Serial tissue sections (less than 10 microns) were taken from the level of the preoptic/anterior hypothalamus through the paraventricular hypothalamus. In situ hybridizations were conducted at room temperature in hybridization buffer. Neurons immunoreactive with antiserum raised against somatostatin were identified in alternate sections using standard immunocytochemical procedures. The anatomical location of the hybridization signal was determined by autoradiography. Our results show that the peri- and paraventricular hypothalamus is rich in transcripts putatively coding for somatostatin and that these transcripts are co-distributed with neurons immunoreactive with antisomatostatin immunoglobulin.

Animals↗
PubMed1992

Application of a fluorescent dye to study connectivity between third ventricular preoptic area grafts and host hypothalamus.

The mutant hypogonadal (hpg) mouse lacks a functioning gene for the neurohormone gonadotropin releasing hormone (GnRH). Previous studies from our laboratory had indicated that the initiation and maintenance of reproductive function in these mice could be brought about by the implantation of normal fetal grafts into adult hosts. Testicular or ovarian growth and other indicators of normal neurosecretory output were always accompanied by survival of GnRH neurons and growth of GnRH axons into the host median eminence where such axons terminate on the hypophysial portal capillaries. To determine if other connections exist between graft and the host hypothalamus, small crystals of the carbocyanine dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (DiI) were applied to either graft or host after fixation of the brain. Tissue sections were analyzed for retrograde and and anterograde movement of the dye. When crystals were placed on the graft, labeled axons were found in the host median eminence or in the host hypothalamus taking an arching trajectory toward the median eminence. Retrogradely labeled neurons in the host were few in number and largely confined to the host arcuate nucleus. With DiI crystals applied to the basal hypothalamus, labeled axons were distributed widely in the host but much sparser in the graft. Axons appeared to enter primarily at sites where the graft and host interface lacked an ependymal lining. Small numbers of retrogradely labeled neurons were also seen in the graft. Most were cells of very simple morphology and were distributed randomly in the graft. When double label experiments were carried out most DiI positive cells in the graft contained GnRH. These results indicate the connectivity between host hypothalamus and the third ventricular preoptic area grafts exists but is limited in nature.

Afferent Pathways↗
PubMed1993

Neuropeptide Y-producing neurons of the arcuate nucleus regenerate axons after surgical deafferentation of the mediobasal hypothalamus.

Dorsolateral and ventomedial surgical deafferentiation of the hypothalamus were used to study the capacity of different types of neuropeptide Y-containing axons afferent to the dorsal hypothalamus to regenerate through surgical lesions. The kinetics of the postlesional responses of transected neuropeptide Y-axons was studied on 30-40 microns thick vibratome sections, either (i) by light or electron microscopy after peroxidase immunostaining for neuropeptide Y or (ii) by confocal microscopy after double fluorescence immunostaining for neuropeptide Y and for glial fibrillary acidic protein. The dorsolateral cut was found to sever 2 main pathways containing neuropeptide Y axons located, respectively, below the bed nucleus of the stria terminalis and in the perifornical region. In both regions transected fibers were found to abut onto the surgical lesion, but even 45 days after the lesion, they were very rarely observed to penetrate into the astroglial scar forming along the lesion. The ventromedial cut was found to sever numerous neuropeptide axons that originate in the underlying arcuate nucleus. Seven to 15 days after the lesion neuropeptide Y fibers located below this type of cut presented a dramatic increase in both their numerical density and their immunostaining intensity. With increasing post-surgery times, an increased number of neuropeptide Y fibers was observed to penetrate and to cross the lesional scar formed by densely packed astrocytic processes. Electron microscope observations further demonstrated that 45 days after the lesion, numerous neuropeptide Y-immunoreactive axonal profiles were included in the scar matrix, which appeared to be mainly composed of closely interdigitating astrocytic processes containing dense bundles of filaments. These data indicate that, in contrast to other neuropeptide Y neurons innervating the dorsal hypothalamus, neuropeptide Y neurons of the arcuate nucleus regenerate axons through the astroglial scar produced by a surgical lesion placed in the ventromedial hypothalamus.

Animals↗
PubMed1996

Somatotropin release-inhibiting factor and galanin innervation in the hypothalamus and pituitary of seabream (Sparus aurata).

The distribution of galanin (GAL) and somatotropin-release-inhibiting-factor (SRIF) immunoreactivity in the hypothalamus and pituitary of the sea bream (Sparus aurata) was studied by immunocytochemistry. An extensive system of neurons immunoreactive with antisera to the two peptides was identified throughout the brain with staining particularly in the hypothalamus. In the hypothalamus, GAL immunoreactive perikarya were detected principally in the nucleus preopticus and nucleus tuberis. Major nerve tracts were observed to sweep down from the hypothalamic nuclei and reached the pituitary via the preoptico-hypophysial tract. Many of the fibers had varicose swellings indicating they were secretory. SRIF immunoreactivity was distributed similarly to GAL but the network of nerve fibers was less dense; no colocalization of these two peptides was seen. SRIF immunoreactive perikarya were present in the preoptic nucleus, the tuberal nucleus, and the basolateral hypothalamus. These perikarya were large and densely staining and were predominately bipolar, although some multipolar perikarya were observed. In the pituitary GAL and SRIF immunoreactivities were confined principally to the pars distalis where fibers infiltrated between growth hormone, prolactin, and adrenocorticotrophic cells. More of the fibers were immunoreactive for SRIF than for GAL. There was no immunoreaction for GAL or SRIF in any of the pituitary cells. There is thus morphological evidence for a neuroendocrine control of the pars distalis by GAL and SRIF and for a possible functional interaction between these two systems.

Animals↗
PubMed1999

Conjugated estradiol increases female sexual receptivity in response to oxytocin infused into the medial preoptic area and medial basal hypothalamus.

The ovarian steroid estradiol (E) has been found to increase both receptor affinity and release of the neuropeptide oxytocin (OT) in plasma membrane preparations. Therefore, we hypothesized that E conjugated to bovine serum albumin at position 6 (E-6-BSA) would increase behavioral responsiveness to OT. Preliminary results showed that 200 ng/microl of E-6-BSA increased sexual receptivity slightly, but not significantly. Therefore, this dose was used as a subthreshold dose to test whether it would increase sexual responsiveness when infused in combination with 100 ng/microl OT. After recovery from cannula implantation surgery animals were injected with 0.5 microg E benzoate daily for 3 days before testing. On the fourth day, after a baseline preinfusion test rats were infused bilaterally with E-6-BSA alone or with OT, OT with BSA, or conjugated progesterone, luteinizing hormone-releasing hormone equimolar to OT alone, or with E-6-BSA or conjugated progesterone alone. When infused into either the medial preoptic area-anterior hypothalamus or the medial basal hypothalamus the combination of OT and E-6-BSA significantly increased sexual receptivity over receptivity after artificial cerebrospinal fluid control infusions. Neither bilateral infusions of OT in combination with conjugated progesterone nor E-6-BSA in combination with luteinizing hormone-releasing hormone enhanced sexual receptivity. Results presented here strongly support the conclusion that some of the effects that E has in sensitizing brain systems to the facilitating effects of OT occur at the membrane level in the medial preoptic area-anterior hypothalamus and medial basal hypothalamus.

Animals↗
PubMed2000

Ultrastructural localization of the receptor for leptin in the rat hypothalamus.

Ultrastructural localization of the leptin receptor in the rat hypothalamus was studied by immunocytochemistry. The antiserum against the leptin receptor which was used specifically recognized the carboxy terminal of the cytoplasmic domain. Intense leptin receptor immunoreactivity was detected in the arcuate, paraventricular, and ventromedial nuclei of the hypothalamus and in the lateral hypothalamic area. At the ultrastructural level, leptin receptor-like immunoreactivity appeared to be concentrated predominantly in perikarya and dendrites of these areas and strong immunolabeling for the leptin receptor was detected in the plasma membrane, rough endoplasmic reticulum, Golgi apparatus, and cytoplasmic matrix. This study provides the first detailed fine structure of leptin receptor-immunoreactive neurons in the rat hypothalamus. It may help to provide better understanding of the functions of leptin in the rat hypothalamus.

Animals↗