Light and electron microscopic immunocytochemistry of GRF-like immunoreactive neurons and terminals in the rat hypothalamic arcuate nuclues and median eminence

Growth hormone-releasing factor (GRF) synthesizing neuronal perikarya and terminals were investigated by light and electron microscopic immunocytochemistry using rat hypothalamus. Immunoreactive neuronal perikarya were located mainly in the ventrolateral part of the arcuate nucleus. They contained well developed cell organella such as mitochondria and rough surfaced endoplasmic reticulum with some expansion. They also contained immunoreactive dense granules (80-120 nm in diameter). On the surface of the immunoreactive neuronal perikarya were frequently found non-immunoreactive axo-somatic synapses. Therefore, the GRF-like immunoreactive neurons were assumed to receive neuronal inputs from other neurons on their neuronal soma. In the external layer of the median eminence large numbers of immunoreactive terminals were distributed particularly around the capillaries of the portal vessel. Electron microscopic immunocytochemistry revealed large numbers of immunoreactive terminals containing immunoreactive dense granules, synaptic vesicles and mitochondria in the vicinity of the basement membrane of the pericapillary space of the portal vessel. Therefore, we concluded that GRF-like immunoreactive substances are released into the portal capillaries from the nerve terminals, which originate from the neuronal perikarya in the ventrolateral part of the arcuate nucleus, and act on growth hormone release in the anterior pituitary. We also suggest that GRF-like immunoreactive neurons have abundant terminal arborization in the external layer of the median eminence.     

Light and electron microscopic immunocytochemistry of β-endorphin/β-LPH-like immunoreactive neurons in the arcuate nucleus and surrounding areas of the rat hypothalamus

β-Endorphin/β-LPH-like immunoreactive neurons in the hypothalamic arcuate nucleus and its surrounding areas were visualized by light and electron microscopic immunocytochemistry. Immunoreactive processes were found in the vicinity of the pia mater, in the lateral part of the external layer of the median eminence and near the lateral wall of the third ventricle. Neuronal perikarya contained immunoreactive dense granules as well as developed cell organellae. They received neuronal inputs from other neurons through axoplasmic and axodendritic synapses. Immunoreactive neuronal processes containing dense granules and mitochondria were found as preterminal elements on non-immunoreactive neuronal soma and dendrites. Immunoreactive processes also make intimate contact with capillaries in the arcuate nucleus near the median eminence.     

Some cellular characteristics of somatostatin neurons and terminals in the periventricular nucleus of the rat hypothalamus and median eminence. Electron microscopic immunohistochemistry

Somatostatin neuronal perikarya and their processes, presumably dendrites, in the periventricular nucleus of the rat hypothalamus and terminals in the median eminence were observed by electron microscopic immunohistochemistry. Neuronal perikarya and processes contained immunoreactive dense granules (100-120 nm in diameter) and other cellular components such as polysomes, rER membranes occasionally showed high electron density. Few axo-somatic terminals were found on the somatostatin neurons, but we could detect a number of preterminal axons on immunoreactive processes, presumably dendrites. Therefore, we considered that somatostatin neurons receive mainly neuronal input through axo-dendritic synapses rather than through axo-somatic ones. In the somatostatin terminals in the external layer of the median eminence immunoreactivity was completely restricted on the granules.     

Immunocytochemical demonstration of dynorphin(PH-8P)-like immunoreactive elements in the human hypothalamus

PH-8P (dynorphin[1-8])-like immunoreactive neuronal perikarya, processes, and terminals located within the human hypothalamus were investigated by the avidin-biotin peroxidase complex (ABC) immunocytochemical procedure. Immunopositive neurons were distributed throughout the hypothalamus. The distributional pattern was found to be similar to that in other mammalian species by the use of antisera against dynorphin. A large number of immunoreactive neuronal perikarya were detected in the supraoptic nucleus (SON) and the magnocellular portion of the paraventricular nucleus (PVN). Their processes appeared to project to the posterior pituitary via the internal layer of the median eminence and their distribution seemed to be less dense than in other mammalian species. PH-8P and vasopressin were colocalized in the neuronal perikarya in the human SON unlike the colocalization of these peptides in the rat SON and PVN. There were a few immunoreactive terminals in the external layer of the median eminence; their immunoreactive substances may be released into the portal veins to act on anterior pituitary cells. In addition, PH-8P-like immunoreactive neurons in the human hypothalamus may project to the extrahypothalamic area.     

Localization of Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) in the Hypothalamus-Pituitary System in Rats: Light and Electron Microscopic Immunocytochemical Studies

The localization of pituitary adenylate cyclase-activating polypeptide (PACAP) in the hypothalamus-pituitary system in rats was examined in light and electron microscopic immunocytochemistry using a specific antiserum to synthetic PACAP 1–38 (R0831). In light microscopic study, intensely PACAP-immunostained perikarya were observed in the supraoptic and paraventricular magnocellular nucleus in the hypothalamus. In the median eminence, many immunoreactive nerve fibers were observed in the internal layer, but a few immunoreactive terminals were noticed in the external layer. In the pituitary gland, numerous immunoreactive nerve fibers were observed in the posterior lobe. In the intermediate lobe, moderately immunostained cells were observed, but in the anterior lobe no immunostained cells were noticed. In electron microscopic study, PACAP-immunoreactivity was examined by avidin-biotin peroxidase complex method. In the perikarya of the supraoptic and paraventricular magnocellular nucleus, DAB-reaction products were distributed diffusely in the cytoplasmic matrix, frequently attaching to the rough-surfaced endoplasmic reticulum. In the nerve terminals of the posterior lobe, reaction products were observed among the secretory granules, but sometimes upon them. In the cells of the intermediate lobe, reaction products were also distributed in the cytoplasmic matrix.     

Immunocytochemical distribution of [Met]enkephalin-Arg-Gly-Leu immunoreactivity in the rat diencephalon

Recently, [Met]Enkephalin-Arg-Gly-Leu (MEAGL) was isolated from bovine adrenal glands, and it was found to be derived exclusively from proenkephalin. Therefore, we investigated the distribution of MEAGL-like immunoreactive neuronal perikarya and fibers in the rat diencephalon pretreated with colchicine by PAP immunocytochemistry. In the thalamus MEAGL immunoreactive neuronal perikarya were distributed in the paraventricular nucleus and the ventral part of the lateral geniculate nucleus. Immunoreactive fibers were found in the paraventricular, paracentral, anteroventral, reuniens and rhomboid nuclei. In addition, immunoreactive fibers were also noted in the anterior pretectal nucleus. In the hypothalamus, immunoreactive neuronal perikarya were observed in the medial preoptic area, anterior and lateral hypothalamic nuclei, perifornical region, parvocellular and postero-magnocellular regions of paraventricular nucleus, ventromedial nucleus, dorsomedial nucleus, arcuate nucleus, premammillary, medial mammillary and lateral mammillary nuclei. The distribution of immunoreactive fibers was similar to that of neuronal perikarya. However, immunoreactive fibers were also observed in the supraoptic and suprachiasmatic nuclei where no immunoreactive neuronal perikarya were detected. Numerous immunoreactive fibers were detected in the external layer of the median eminence, but there were few in the internal layer. The similarity and difference in the distribution between MEAGL and other proenkephalin peptides such as [Met]enkephalin were also discussed.     

GRF immunoreactive neurons in the paraventricular nucleus of the rat: an immunohistochemical study with monoclonal and polyclonal antibodies

Our study demonstrates a complex GRF neuronal system within the rat hypothalamus. Using both high affinity polyclonal and high specificity monoclonal antibodies to rat (r) GRF, we have substantiated evidence for immunoreactive GRF (GRF-i) perikarya in the parvocellular portion of the paraventricular nucleus. Other hypothalamic areas containing rGRF-positive perikarya include the lateral arcuate nucleus, lateral hypothalamus, perifornical area and dorsomedial nucleus. GRF-i neuronal terminals were seen in the external zone of the median eminence, more rostrally in the periventricular nucleus, and near the suprachiasmatic nucleus and more caudally in the dorsomedial nucleus and ventral premammillary nucleus.     

Neurotensin in the rat median eminence: the possible sources of neurotensin-like fibers and varicosities in the external layer

The possible sources of neurotensin-like immunoreactive axons in the median eminence were studied after several experimental surgical approaches including unilateral lateral retrochiasmatic area transection, midsagittal knife cut through the median eminence, complete surgical isolation of the medial basal hypothalamus and bilateral paraventricular nucleus lesions. Both immunohistochemical and radioimmunoassay data demonstrate that neurotensin-containing neuronal located in the hypothalamic arcuate nuclei represent the main source of neurotensin occurring in the external zone of the median eminence of the rat: (1) neither the complete isolation of the medial basal hypothalamus nor the transection of the major neuronal input channel to the median eminence in the lateral retrochiasmatic area altered neurotensin-like immunoreactivity in the median eminence; (2) bilateral lesioning of the paraventricular nucleus resulted in insignificant changes of neurotensin level in the median eminence; and (3) two days after lesioning the median eminence an increased amount of retrogradely accumulated neurotensin-like immunoreactivity was found in several perikarya of the arcuate nuclei due to the blockage of axonal transport in the transected fibers. Retrograde accumulation of neurotensin-like material in other cells scattered in the anterior hypothalamus (in the paraventricular, paraventricular and anterior hypothalamic nuclei) indicates that in addition to the arcuate neurons these neurons may also participate in the neurotensin innervation of the median eminence.     

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.     

Light and electron microscopic study of somatostatin-like immunoreactive neurons in rat hippocampus

The distribution and fine structure of somatostatin-like immunoreactive neurons in rat hippocampus and gyrus dentatus were investigated by light and electron microscopic immunocytochemistry. Somatostatin-like immunoreactive neuronal perikarya and fibers could be visualized by using modified PAP immunocytochemistry. Immunoreactive neurons were selectively localized in the stratum orience of the hippocampus and hilar region of the gyrus dentatus, however, immunoreactive neurons were also sparsely observed throughout hippocampal formation. Somatostatin-like immunoreactive varicosities were abundantly distributed in the stratum pyramidale and some of them were considered to terminate on the pyramidal cells. Somatostatin-like immunoreactive neurons in the hippocampal formation generally contained many mitochondria, well-developed rough surfaced endoplasmic reticulum (rER), polysomes and some dense granules. Immunoreactivity was observed especially in the dense granules and membrane of rER. Pre- and post-synaptic elements of somatostatin-like immunoreactive neurons were detected throughout the hippocampal formation.     

Substance P-like immunoreactive neurons in the arcuate nucleus project to the median eminence in rat

Substance P-containing nerve fibers in the median eminence of rat arise in cells located in the arcuate nucleus. Two days following surgical lesioning of the median eminence immunoreactive substance P accumulated in neuronal perikarya in the middle part (rostrocaudally) of the arcuate nucleus, mainly in its ventromedial portion. Substance P-immunostained cells appeared nowhere else in the hypothalamus following surgical lesion of the median eminence while they were found in several hypothalamic and extrahypothalamic cell groups after colchicine treatment.     

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.     

Immunohistochemical mapping of neuropeptides in the premamillary region of the hypothalamus in rats

The topographical distribution of neuropeptide-containing cell bodies, fibers and terminals was studied in the premamillary region of the rat hypothalamus using light microscopic immunohistochemistry. Alternate coronal sections through the posterior third of the hypothalamus of normal and colchicine-treated male rats were immunostained for 19 different neuropeptides and their distributions were mapped throughout the following structures: the ventral and dorsal premamillary, the supramamillary, the tuberomamillary and the posterior hypothalamic nuclei, as well as the premamillary portion of the arcuate nucleus and the postinfundibular median eminence. Seventeen of the investigated neuropeptides were present in neuronal perikarya, nerve fibers and terminals while the gonadotropin associated peptide and vasopressin occurred only in fibers andn terminals. Growth hormone-releasing hormone-, somatostatin-, α-melanocyte stimulating hormone-, adrenocorticotropin, β-endorphin- and neuropeptide Y-immunoreactive neurons were seen exclusively in the premamillary portion of the arcuate nucleus. Thyrotropin-releasing hormone-, dynorphin A- and galanin-containing neurons were distributed mainly in the arcuate and the tuberomamillary nuclei. A high number of methionine- and leucine-enkephalin-immunoreactive cells were detected in the arcuate and dorsal premamillary nuclei, as well as in the area ventrolateral to the fornix. Substance P-immunoreactive perikarya were present in very high number within the entire region, in particular in the ventral and dorsal premamillary nuclei. Cell bodies labelled with cholecystokinin- and calcitonin gene-related peptide antisera were found predominantly in the supramamillary and the terete nuclei, respectively. Corticotropin-releasing hormone-, vasoactive intestinal polypeptide- and neurotensin-immunoreactive neurons were scattered randomly in low number, mostly in the arcuate and the ventral and dorsal premamillary nuclei. Peptidergic fibers were distributed unevenly throughout the whole region, with each peptide showing an individual distribution pattern. The highest density of immunoreactive fibers was presented in the ventral half of the region including the arcuate, the ventral premamillary and the tuberomamillary nuclei. The supramamillary nucleus showed moderately dense fiber networks, while the dorsal premamillary and the posterior hypothalamic nuclei were poor in peptidergic fibers.     

Immunohistochemical localization of avian pancreatic polypeptide-like immunoreactivity in the rat hypothalamus

The distribution of avian pancreatic polypeptide-like (APP) immunoreactivity within the rat hypothalamus was investigated with the indirect immunoperoxidase method. APP immunoreactive perikarya are found in largest numbers in the retrochiasmatic area, the arcuate nucleus, and the supracommissural portion of the interstitial nucleus of the stria terminalis. Small clusters of immunoreactive neurons are also consistently observed in the ventral aspect of the medial preoptic area and lateral hypothalamic area, immediately dorsolateral to the optic chiasm and tracts. These neurons are apparent in all animals but are more intensely strained and occur in larger numbers following colchicine pretreatment. Other immunoreactive neurons are visible only in colchine-treated rats and are scattered throughout the anterior and lateral hypothalamic areas and the supramammillary nucleus. Immunoreactive axons and terminal fields present an extensive and highly characteristic distribution throughout the hypothalamus, which in many instances exhibits differential distribution within specific subfields of hypothalamic nuclei and areas. The heaviest concentrations of APP immunoreactive axons are present in the periventricular nucleus throughout the rostrocaudal extent of the hypothalamus, the ventrolateral portion of the suprachiasmatic nucleus, the retrochiasmatic area, the parvocellular paraventricular nucleus, the ventral supraoptic nucleus, the perifornical nucleus, the ventral dorsomedial nucleus, and the arcuate nucleus. Moderate plexuses of immunoreactive fibers are also present in the medial preoptic area, the anterior and lateral hypothalamic areas, the nucleus circularis, the median eminence, and the ventral premammillary area. Other areas, such as the ventromedial nucleus, contain virtually no immunoreactive axons but are encapsulated by a dense plexus of immunoreactive terminals. The distribution of a major component of APP immunoreactive fibers exhibits a marked similarity to that of previously described norepinephrine-containing hypothalamic afferents. Other groups of APP immunoreactive perikarya and fibers appear to represent components of intrinsic diencephalic systems.     

Monosynaptic Pathway Between the Arcuate Nucleus Expressing Glial Type II Iodothyronine 5'-Deiodinase mRNA and the Median Eminence-Projective TRH Cells of the Rat Paraventricular Nucleus

Recent evidence suggests that the thyroid regulation of thyrotropin-releasing hormone (TRH)-containing neurons in the paraventricular nucleus of the hypothalamus involves the activation of other hypothalamic neural circuits. For example, the arcuate nucleus and not the paraventricular nucleus contains the highest enzyme activity of 5′-deiodinase type II, an enzyme that is pivotal for the local synthesis of T3. This experiment was undertaken to demonstrate whether a monosynaptic pathway exists between the arcuate nucleus and those TRH cells of the paraventricular nucleus that are neuroendocrine, i.e. project to the external layer of the median eminence. A specific cRNA probe derived from the coding region of deiodinase type II was used for the in situ hybridization histochemistry which was combined with immunocytochemistry for a specific marker of glial cells, glial fibrillary acidic protein (GFAP). The hybridization signals were present within the hypothalamus in the arcuate nucleus–median eminence region and in the periventricular area. The periventricular labeling was localized to the ependymal layer of the third ventricle and no hybridization product was detected in the paraventricular nucleus and other hypothalamic nuclei adjacent to the third ventricle. Within the median eminence, numerous cells containing the hybridization product were located in the internal layer adjacent to the floor of the third ventricle and in the external layer adjacent to the surface of the brain. In the dorso- and ventromedial regions of the arcuate nucleus, deiodinase type II mRNA-containing cells were also detected. Numerous type II deiodinase mRNA-containing cells in the median eminence and arcuate nucleus were also found to be immunopositive for GFAP. The abundance of arcuate cells expressing the hybridization product was lower than those in the periventricular region or in the median eminence. The anterograde tracer, Phaseolus vulgaris leucoagglutinin, was injected into the medial parts of the arcuate nucleus where the in situ hybridization experiment detected deiodinase type II mRNA. Simultaneously with the anterograde tracing, the retrograde tracer, Fluoro-Gold, was injected into either the median eminence or the general circulation. Light and electron microscopic double and triple immunolabeling experiments on vibratome sections of colchicine-pretreated animals revealed that arcuate fibers innervate TRH cells within the parvicellular region of the paraventricular nucleus. Populations of these TRH cells receiving afferents from the arcuate nucleus were also retrogradely labelled from either the median eminence or the general circulation indicating their direct role in the regulation of thyrotropin secretion from the anterior pituitary. The majority of arcuate nucleus efferents on TRH cells were found to establish symmetrical synaptic connections. The present results provided direct evidence of a monosynaptic pathway between the hypothalamic site of local thyroid hormone production, the arcuate nucleus, and neuroendocrine TRH cells in the paraventricular nucleus. This signalling modality may play an important role in thyroid feedback on TRH cells. Since the arcuate nucleus is involved in the regulation of central mechanisms controlling diverse homeostatic functions, including reproduction and feeding, the pathway described in this study may also carry integrated signals related to reproduction and ingestion to TRH-producing cells.     

Vasoactive intestinal peptide (VIP) in magnocellular neurons of the hypothalamo-neurohypophysial system of the mink (Mustela vision) is co-localized with vasopressin or oxytocin.

The distribution of vasoactive intestinal peptide (VIP) was analysed in perikarya of the mink hypothalamus with immunohistochemistry and, surprisingly, a large population of magnocellular VIP-immunoreactive neurons was present in the paraventricular and supraoptic nuclei as well as in accessory hypothalamic nuclei. From perikarya in the paraventricular as well as supraoptic nuclei, a large number of VIP immunoreactive nerve fibers was observed to enter the hypothalamo-neurohypophysial tract. Within the median eminence, a high density of VIP-immunoreactive nerve fibers was present in the external and internal zones. Fibers in the external zone of the median eminence were endowed with varicosities and perivascular terminals, while fibers in the internal zone were smooth and without terminal specializations. From the internal zone of the median eminence, fibers coursed via the infundibular stalk to terminate in perivascularly situated terminals in the neurohypophysis. In addition, a substantial number of small VIP-immunoreactive perikarya was observed within the suprachiasmatic nucleus. These perikarya were immunoreactive to neither vasopressin nor neurophysin. To elucidate the co-existence of VIP-immunoreactivity with vasopressin, oxytocin or neurophysin, a sequential double immunoperoxidase procedure to localize antigens with diaminobenzidine and benzidine dihydrochloride as chromagens was performed. From these experiments it was evident that VIP in nearly all magnocellular hypothalamo-neurohypophysial neurons co-existed with neurophysin. Based on a semi-quantitative estimate, half the VIP-immunoreactive magnocellular perikarya co-stored vasopressin, while another half co-stored oxytoxin. The present study describes the presence of a large population of VIP-containing neurons in the hypothalamo-neurohypophysial system of the mink. These findings raise evidence that within the mink, VIP may be involved in neurohypophysial physiology.     

Distribution of immunoreactive substance P neurons in the hypothalamus and pituitary of the rhesus monkey

The distribution of immunoreactive substance P (IR-SP) neurons was examined in the hypothalamus and pituitary gland of the rhesus monkey by using the peroxidase-antiperoxidase immunocytochemical technique. Immunoreactive SP cell bodies were observed in the arcuate nucleus, in the region lateral to the arcuate nucleus, and in the median eminence (ME). Immunoreactive SP cells were also seen in the periventricular area of the dorsal tuberal region. A rich network of SP fibers was concentrated in the arcuate region, and the fiber stain was particularly dense in the external zone of the median eminence and in the external layer of the infundibular stalk. Also, substance P fibers were seen in the internal layer of the pituitary stalk and in the neural lobe of the pituitary gland. Outside the hypothalamus a dense network of IR-SP fibers was observed in the globus pallidus.     

Electron microscopic immunocytochemical evidence for the existence of bidirectional synaptic connections between growth hormone-releasing hormone- and somatostatin-containing neurons in the hypothalamus of the rat

Synaptic contacts between growth hormone-releasing hormone (GHRH)- and somatostatin-containing neurons were demonstrated in the rat hypothalamus by a double-staining immunocytochemical method at the electron microscopic level. Somatostatin-immunoreactive nerve terminals synapse on GHRH-positive dendrites and cell bodies in the arcuate nucleus. A fine network of GHRH-immunopositive nerve terminals was observed at the light microscopic level in the rostral part of the periventricular nucleus and in the dorsal part of the arcuate nucleus around somatostatin-containing neuronal elements. With the electron microscope synaptic contact between GHRH-containing nerve terminals and somatostatin-containing dendrites are demonstrated. The reciprocal innervation between GHRH- and somatostatin-containing neurons that project to the median eminence and regulate growth hormone secretion must allow them to coordinate their activities.     

The fine structural organization of tyrosine hydroxylase immunoreactive neurons in rat arcuate nucleus

The fine structure of the tyrosine hydroxylase (TH) immunoreactive neurons of the hypothalamic arcuate nucleus was examined by means of immunocytochemistry [peroxidase-antiperoxidase (PAP) method], utilizing an antibody against TH. Immunolabeled axon terminals were observed infrequently and were located predominantly in the lateral region, whereas numerous labeled perikarya and dendrites were found throughout the nucleus. The labeled terminals, containing primarily clear and occasionally dense core vesicles, were never observed in synaptic contact. On the other hand, unlabeled axon terminals were frequently seen synapsing on labeled dendrites. In addition, the labeled dendrites were often seen in direct apposition to other neuronal elements such as both labeled and unlabeled perikarya. In contrast, unlabeled dendrites were never seen apposed to labeled perikarya. Labeled dendrites also occurred in direct contact with one another and with unlabeled dendrites. Moreover, numerous labeled dendrites were encountered along tanycytic processes. Dendrites engaged in tanycytic appositions were occasionally partially encompassed by thin sheaths emanating from the tanycytic process. The extensive contact made by the labeled dendritic profiles on both labeled perikarya and dendrites suggests that tubero-infundibular dopaminergic (TIDA) cells may communicate with each other by means of dendritic release of dopamine. The presence of appositions between labeled dendrites and both unlabeled perikarya and dendrites suggests that the TIDA system also influences other neuronal populations through its dendrites. Finally, the dendrotanycytic relationship suggests that the TIDA system may play some role in the regulation of tanycytic function.     

The fine structures of the VIP-Like immunoreactive neurons in the cat hypothalamus

The fine structures of the VIP-like immunoreactive neurons in the suprachiasmatic nucleus (SCN) and the arcuate nucleus ( ARN ) of the cat hypothalamus were investigated by electron microscopic immunocytochemistry. The VIP-like immunoreactive soma and fibers could be successfully visualized by a modified PAP method. VIP-like immunoreactive neurons in both nuclei contained immunoreactive rER, Golgi complexes and many immunoreactive granules, as well as well developed mitochondria. VIP-like immunoreactive synaptic endings with synaptic membrane specialization of Gray's type I and II were found in the SCN. Moreover VIP-like immunoreactive preterminal elements that made synaptic contact with VIP-like immunoreactive neuronal soma were also detected. On the other hand, it was difficult to detect typical preterminal endings with immunoreactivity in the ARN ; however, VIP-like immunoreactive processes in contact with the basement membrane of the capillaries were observed. These observations indicate that VIP-like immunoreactive neurons in the SCN act as intrinsic neurons and are involved in neuroendocrine function in ARN .