Dynamic aspects of the LHRH system associated with ovulation in the little brown bat (Myotis lucifugus)

Adult female bats were collected from natural roosting sites in pre-ovulatory and post-ovulatory conditions. LHRH neurones of these animals were examined using light and electron microscopic immunocytochemistry, and LHRH tissue contents were measured by radioimmunoassay. Comparisons between the two groups of bats revealed that the number of LHRH perikarya detected immunocytochemically, as well as hypothalamic LHRH content, were significantly reduced in post-ovulatory animals. Distributions of immunoreactive perikarya were, however, strikingly similar in both groups. The reduction in immunoreactive cell number observed after ovulation was therefore not restricted to an anatomically defined subset of neurones, but was evident throughout the population. The projection of LHRH-immunoreactive fibres that extend into the pituitary neural lobe in this species also exhibited changes related to endocrine condition. Morphometric indices of fibre density in the neural lobe were significantly reduced in post-ovulatory bats, as was LHRH content of the lower infundibular stalk and neural lobe. Fine structural study of perikarya revealed complex anatomical interactions between LHRH-immunopositive elements, especially in post-ovulatory bats. These interactions included direct apposition of perikarya, as well as more elaborate networks involving various combinations of perikarya and large- and small-caliber processes. These changes in the LHRH system associated with ovulation suggest reduction of stored peptide within perikarya and depletion from terminals within the lower infundibular stem and neural lobe. Parallel reductions in hypothalamic and neural lobe LHRH content during the periovulatory period support the hypothesis that the neural lobe component of the system contributes to control of gonadotrophin secretion in this species. Finally, increased complexity of anatomical contact between components of the LHRH system may be related to activation of this cell population in spring.     

Immunocytochemical localization of neuropeptides in the hypothalamus of the Japanese long-fingered bat,Miniopterus schreibersii fuliginosus

Summary To elucidate the role of hypothalamic neuropeptides in regulation of reproductive phenomena of seasonally breeding feral mammals, we used Japanese long-fingered bats, Miniopterus schreibersii fuliginosus, for immunocytochemical study of distribution of the following neuropeptides in the hypothalamus: arginin vasopressin, oxytocin, luteinizing hormone-releasing hormone, somatostatin, corticotropin-releasing factor, and growth hormone-releasing factor. The size, shape and location of supraoptic, paraventricular, suprachiasmatic, and arcuate nuclei of the bat were determined. Arginin vasopressin-and oxytocin-immunoreactive magnocellular neurons were found in the supraoptic and paraventricular nuclei, where they exhibited separate distribution into two distinct groups. Parvocellular arginin vasopressin neurons occurred only in the suprachiasmatic nucleus. The hibernating bats exhibited slightly increased numbers of vasopressin and oxytocin neurons in the supraoptic and paraventricular nuclei. The pregnant bat displayed further increased numbers of vasopressin and oxytocin neurons in both nuclei. Somatostatin-immunoreactive neurons in the paraventricular nucleus were also immunopositive to anti-oxytocin serum, while those in the ventromedial and arcuate nuclei reacted solely to anti-somatostatin serum. They projected to the anterior median eminence and infundibular stalk. Luteinizing hormone-releasing hormone-immunoreactive perikarya were scattered throughout the basal hypothalamus, being particularly abundant in the arcuate nucleus. They were larger in size in hibernating bats than those in normal (non-pregnant) and pregnant females. They projected fibers mainly to the internal layer of the median eminence and infundibular stalk. A few luteinizing hormone-releasing hormone-reactive fibers were also observed in the organum vasculosum laminae terminalis, lateral habenular nuclei, pineal stalk, retroflexus fasciculus, and olfactory tubercle. Corticotropin releasing factor-immunoreactive perikarya were distributed in the paraventricular nucleus and medial preoptic area and projected into the external layer of the anterior median eminence, while growth hormone-releasing factor-immunoreactive perikarya occurred only in the arcuate nucleus and projected into the posterior part of the median eminence.     

Immunocytochemical study of the GABAergic innervation of the mouse pituitary by use of antibodies against gamma-aminobutyric acid (GABA)

Summary The GABAergic innervation of the mouse pituitary, including the median eminence, was studied at light microscopic and ultrastructural levels by use of a pre-embedding immunocytochemical technique with antibodies directed against GABA. In the median eminence, a high density of GABA-immunoreactive fibers was found in the external layer where the GABAergic varicosities were frequently observed surrounding the blood vessels of the primary capillary plexus. In the internal and subependymal layers, only few fibers were immunoreactive. The intense labeling of the external layer was observed in the entire rostro-caudal extent of the median eminence. In the pituitary proper, a dense network of GABA-immunoreactive fibers was revealed throughout the neural and intermediate lobes, entering via the hypophyseal stalk. The anterior and tuberal lobes were devoid of any immunoreactivity. The GABA-immunoreactive terminals were characterized in the median eminence, and in the intermediate and posterior lobes at the electron-microscopic level. They contained small clear vesicles, occasionally associated with dense-core vesicles or neurosecretory granules. In the intermediate lobe they were seen to be in contact with the glandular cells. In the posterior lobe and in the median eminence, GABA-immunoreactive terminals were frequently located in the vicinity of blood vessels. These results further support the concept of a role of GABA in the regulation of hypophyseal functions, via the portal blood for the anterior lobe, directly on the cells in the intermediate lobe, and via axo-axonic mechanisms in the median eminence and posterior lobe.     

Relationship of the central ACTH-immunoreactive opiocortin system to the median eminence and the pituitary gland of the rat

Summary By immunocytochemical methods, the present study describes ACTH-immunoreactive fibers in the pituitary stalk and neural lobe. This opiocortin-hypothalamo-neural lobe projection arises in a bed nucleus of perikarya in the basal hypothalamus, follows supraoptico-hypophyseal fibers in the zona interna of the median eminence, and distributes throughout the neural lobe. No ACTH-immunoreactive fibers project to the zona externa; some are present in the subependymal layer and at the lateral margins of the median eminence. Further studies must identify the role of these fibers in posterior lobe function. It remains also to be determined whether this system terminates upon primary pituitary portal capillaries and delivers opiocortin neuropeptides to the adenohypophysis.Supported by NIH Grants HD-07962, NS-15345 and AM-22029The skillful technical assistance of Donna Wilson, Nancy Dembs and Jay Hocton is thankfully acknowledged     

Melanin-concentrating hormone stimulates the release of luteinizing hormone-releasing hormone and gonadotropins in the female rat acting at both median eminence and pituitary levels

The purpose of this study was to investigate whether melanin-concentrating hormone (MCH) acts directly on the median eminence and on the anterior pituitary of female rats regulating LHRH and gonadotropin release. In addition, immunohistochemistry was used to examine the density and distribution of MCH-immunoreactive fibers in the median eminence of proestrous rats. MCH-immunoreactive fibers were found in both the internal and external layers of the median eminence and in close association with hypophysial portal vessels. In the first series of in vitro experiments, median eminences and anterior pituitaries were incubated in Krebs-Ringer bicarbonate buffer containing two MCH concentrations (10(-10) and 10(-8) M). The lowest MCH concentration (10(-10) M) increased (P < 0.01) LHRH release only from proestrous median eminences. Anterior pituitaries incubated with both MCH concentrations also showed that 10(-10) M MCH increased gonadotropin release only from proestrous pituitaries. In the second series of experiments, median eminences and pituitaries from proestrous rats were incubated with graded concentrations of MCH. MCH (10(-10) and 10(-9) M) increased (P < 0.01) LHRH release from the median eminence, and only 10(-10) M MCH increased (P < 0.01) LH and FSH release from the anterior pituitary. The effect of MCH on the stimulation of both gonadotropins from proestrous pituitaries was similar to the effect produced by LHRH. Simultaneous incubation of pituitaries with MCH and LHRH did not modify LH but increased the FSH release induced by LHRH. The present results suggest that MCH could be involved in the regulation of preovulatory gonadotropin secretion.     

Relationship between neuropeptide Y and luteinizing-hormone-releasing hormone immunoreactivities in the hypothalamus and preoptic region

The purpose of this study was to examine the anatomical relationships of perikarya and fibers containing neuropeptide Y (NPY) and luteinizing-hormone-releasing hormone (LHRH) in the hypothalamus and preoptic region of female rats. In view of our previous report of stimulatory effects of estrogen on LHRH and NPY levels in the median eminence, animals were bilaterally ovariectomized and subsequently implanted subcutaneously with capsules containing estradiol benzoate in oil or vehicle. Following intracerebroventricular injection of colchicine, rats were perfused with fixative and their brains sectioned and processed for immunohistochemical visualization of NPY and LHRH in the same section and in consecutive sections. Estrogen treatment had no discernible effect on the distribution or relationship of these peptides. NPY-immunoreactive fibers were intimately associated with LHRH-labeled primary dendrites and perikarya in the medial preoptic region and horizontal limb of the diagonal band of Broca. Fibers containing NPY or LHRH overlapped extensively in the lateral palisade region of the median eminence and also in the subependymal and internal zones. The external zone of the median eminence displayed relatively less overlap of these peptide systems. LHRH-immunoreactive axons coursed among NPY-labeled perikarya in the arcuate nucleus and appeared to contact these cells. These results suggest that NPY-containing axons may influence LHRH-positive neurons at the cell body and also at the site of axon termination in the median eminence. LHRH-containing axons appear to contact NPY-immunoreactive perikarya in the arcuate nucleus and may interact with terminals in the median eminence. This arrangement may provide a mechanism for communication between NPY and LHRH neurons and for the neuroendocrine coordination of hypothalamic NPY and LHRH secretion before ovulation.     

Immunohistochemical localization of the luteinizing hormone releasing hormone (LHRH)-containing structures in the central nervous system of the domestic fowl

Summary The location of LHRH-containing neuronal elements was investigated in the domestic fowl by means of immunohistochemical techniques. LHRH antisera were raised against synthetic LHRH in the rabbit. The antiserum used in the present study cross-reacted with LHRH of mammalian and avian tissues.LHRH-immunoreactive perikarya are located in the preoptic and in the septal areas, and in the bulbus olfactorius; however, no LHRH-immuno-reactive perikarya were found in the tuberal part of the hypothalamus. LHRH-immunoreactive fibers course from these areas toward the median eminence mainly along the wall of the third ventricle in the form of a periventricular network. Originating from the same cell groups other fibers run caudally immediately above the optic chiasma, forming the median bundle of the tractus preoptico-infundibularis. The third bundle running toward the OVLT is named the tractus preoptico-terminalis. In addition to these structures, LHRH-containing fibers and terminals were also present in different regions of the limbic system, in the dorsal part of the hippocampus, in the tuberculum and bulbus olfactorius, as well as in the optic lobe, nuclei commissurales tectales, organon subcommissurale, periaqueductal area, and pars ventralis mesencephali.The general distribution of the LHRH system in the chicken corresponds principally to that described previously in rodents (Sétáló et al. 1976, 1978). However, some subtle differences were demonstrated between the location of the LHRH system in birds and mammals.     

Immunohistochemistry of the hypothalamic neuropeptides and anterior pituitary cells in the Japanese quail

The pars distalis of the avian adenohypophysis consists of well-defined cephalic and caudal lobes which are distinct in their cellular constituents. Immunocytochemical investigations on the pituitary hormones of the pars distalis of the Japanese quail reveal five types of secretory cells, adenocorticotropin (ACTH) cells, prolactin (PRL) cells, thyroid-stimulating hormone (TSH) cells, growth hormone GH (STH) cells, and FSH/LH (gonadotropic) cells. The ACTH cells, TSH cells, and PRL cells are restricted to the cephalic lobe, and GH (STH) cells are confined to the caudal lobe, while FSH/LH cells are distributed throughout the cephalic and caudal lobes. The median eminence of birds has distinct anterior and posterior divisions, each with different neuronal components. The avian hypophysial portal vessels also consists of two groups, anterior and posterior. The peculiar arrangement and distribution of the avian hypophysial portal vessels are possibly related to the distribution of neuropeptides in the two divisions of the median eminence and to the cytological and functional differentiation of two lobes of the pars distalis. The localization of perikarya and fibers containing luteinizing hormone releasing hormone (LHRH), somatostatin, vasotocin, mesotocin, corticotropin-releasing factor (CRF), vasoactive intestinal polypeptide (VIP), glucagon, metenkephalin, and substance P in the hypothalamus and median eminence of the Japanese quail has been investigated by means of immunohistochemistry using antisera against the respective neuropeptides. LHRH-, somatostatin-, VIP-, met-enkephalin-, and substance P-immunoreactive fibers are localized in the external layer of the anterior and posterior divisions of the median eminence, while CRF- and vasotocin-reactive fibers are demonstrated only in the external layer of the anterior division of the median eminence. The metenkephalin fibers are thicker in the anterior median eminence but the substance P fibers are more abundant in the posterior division. Mesotocin fibers occur only in the internal layer of the median eminence and neural lobe.     

Studies on hypothalamo-pituitary corticoliberin system. II. Corticotropin-releasing factor (CRF) and neurophysin (NP) immunoreactive neurocytes in the hamster

The aim of the study was to investigate CRF- and neurophysin-immunoreactive neurocytes in hypothalamo-pituitary system of the hamster. CRF-immunoreactive nerve fibers were observed mainly in the outer layer of the median eminence and pituitary stalk and also in the neurohypophysis. On the contrary, neither intermediate lobe nor anterior pituitary contained CRF-immunoassayable substance. The pattern of distribution of neurophysin-immunoreactive fibres was different from CRF-immunoreactive fibres as far as a median eminence, pituitary stalk and neurohypophysis are concerned. Between the tannocytes of the III ventricle and nervous fibres forming the internal layer of the median eminence a CRF- and neurophysin-immunoreactive perikaryons of neurocytes were found. Results of the study suggest regulatory function of CRF-immunoreactive neurons of the hamster hypothalamo-pituitary system in controlling of ACTH secretion. Moreover, the distribution of CRF-immunoreactive substances in hamster hypothalamo-pituitary system shows some peculiarities if compared with other rodents.     

Biosynthesis of LHRH: Inferences from immunocytochemical studies

Inferences regarding biosynthesis of LHRH in rats are made from immunocytochemical studies using LHRH antisera with varied and specific binding requirements. Immunoreactive perikarya were observed with antisera that could bind putative large molecular weight precursors of LHRH. No cells were detected with an antiserum that requires free decapeptide terminals and could not bind extended precursors. No such differential immunoreactivity was apparent in neuronal processes and neurovascular terminals. Features of intracellular processing of LHRH which can be inferred from these immunocytochemical data are: (1) the decapeptide is initially synthesized within neuronal cell bodies as a larger molecular weight peptide, extended at both the N- and C-terminals; (2) processing occurs as the newly synthesized material is transported along neuronal processes; and (3) intermediate molecular forms are converted to the active decapeptide primarily in distal portions of neuronal fibers, including the neurovascular terminal. Immunocytochemical observations in other mammalian species (humans, monkeys, ferrets and bats) allow us to further suggest that the dynamics of maturation of this hormone may differ among mammals.     

Location of neurons projecting to the hypophysial stalk — median eminence in ring doves (Streptopelia roseogrisea)

The median eminence/pituitary stalk represents the final common pathway for fibers from neurons that project to the pituitary gland. We have used the lipophilic fluorescent tracer 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine perchlorate (DiI) to determine the location of neurons projecting to the median eminence/pituitary stalk in ring doves. The tracer can be precisely applied to fixed tissue, in areas to which it is otherwise difficult to gain access. Follwwing application of DiI to the median eminence/pituitary stalk, labeled neurons were detected in six distinct regions: the ventromedial hypothalamic nucleus, paraventricular nucleus, supraoptic nucleus, in and ventral to the lateral forebrain bundle, preoptic area, and lateral septum. Labeled fibers branched extensively in the diencephalon, particularly along the third ventricle and in the septal-preoptic area. Sparse fiber labeling occurred caudal to the tuberal hypothalamus, even though these regions were close to the application site of the tracer. Labeled cerebrospinal-fluid-contacting cells were seen in the paraventricular region of the third ventricle. The results indicate that the avian neuronal system that projects to the median eminence and neural lobe occurs in diffuse clusters largely along the midline region of the hypothalamic septal-preoptic area. The paucity of fiber staining caudal to the tuberal hypothalamic region indicates that cells of these regions do not project to the median eminence/pituitary.     

Light-microscopic immunocytochemical localization of growth hormone-releasing factor in the human hypothalamus

Summary The distribution of growth hormone-releasing factor (GRF)-like immunoreactivity in the human hypothalamus was studied by light-microscopic immunocytochemistry. With antibodies that we developed against synthetic human pancreatic GRF (hpGRF), we localized GRF immunoreactivity in neuronal cell bodies that were observed only in the infundibular (arcuate) nucleus. Immunostained nerve fibers were found in large numbers in the neurovascular zone of the median eminence, in the proximal portion of the pituitary stalk and in periventricular areas. These localizations are in agreement with those of studies recently performed in other species and strongly suggest that GRF can be released into the capillaries of the pituitary portal plexus to reach the anterior pituitary gland. The projections of GRF neurons in extra-infundibular regions suggest that GRF can also act as a neuromodulator or neurotransmitter in the hypothalamus.     

Localization of melanin-concentrating hormone-like immunoreactivity in the brain and pituitary of the frog Rana ridibunda

The distribution of melanin-concentrating hormone (MCH) in the central nervous system of the frog Rana ridibunda was determined by the indirect immunofluorescence technique using antibodies against synthetic salmon MCH, generated in rabbits. The most prominent group of MCH-like containing perikarya was detected in the preoptic nucleus. Comparatively, a moderate number of cell bodies was observed in the dorsal infundibular nucleus and in the ventral thalamic area. Brightly immunofluorescent nerve bundles were found in the preoptic nucleus and in the ventral infundibular nucleus, coursing towards the internal zone of the median eminence and the pituitary stalk. An intense network of immunofluorescent fibers was localized in the neural lobe of the pituitary. The subcellular localization of MCH-like material was studied in the neurohypophysis using the immunogold technique. It was demonstrated that MCH-like material was contained in dense core vesicles (80–90 mm in diameter) within specific nerve terminals. The present findings indicate that, in amphibians, MCH-like peptide is located in specific hypothalamic neurons. Our data suggest that MCH may be released by neurohypophyseal nerve endings as a typical neurohormone.     

Immunoreactive material resembling ovine prolactin in perikarya and nerve terminals of the rat hypothalamus

Summary The presence of prolactin (PRL)-like material is demonstrated in the brain of rats with the aid of anti-ovine PRL (oPRL) IgG as primary antibody in the unlabeled antibody-enzyme method. Immunoreactive deposits are visualized as an intraneuronal constituent with a widespread distribution in the hypothalamus and neural lobe of the pituitary. Dense networks of reactive nerve terminals derived from two prominent fibre tracts, a ventral (VHT) and a dorsal hypothalamo-neurohypophysial tract (DHT) are seen. The VHT is confined to the median eminence and pars oralis tuberis, the DHT to the pars caudalis tuberis. Both fibre tracts pass through the infundibular stalk into the neural lobe. The origin of the immunoreactive nerve terminals can be elucidated only to some extent. The VHT gives off beaded fibres entering the ependymal and glandular layer of the median eminence. Immunoreactive perikarya are observed in the supraoptic nucleus, the paraventricular nucleus, the anterior hypothalamic nucleus, the anterior commissural nucleus, the preoptic nucleus and the interstitial nucleus of the stria terminalis. A few of the immunoreactive perikarya are observed in close connection with brain vessels and the ependymal cells of the third ventricle. The results indicate that the anti-oPRL has a unique region specificity implying that only a segment of the mammalian PRL molecule is present in these nuclei of the brain. Fragments of PRL may function as neuromodulators or neurotransmitters in the rat brain.We are indebted to Dr. Mogens Hammer, Rigshospitalet, Copenhagen for the gift of Arg-VP and anti-VP, and to NIAMDD for the gift of ovine PRL, ratPRL, anti-rPRL, anti-hPRL and bovineSTH     

Distribution of atrial natriuretic factor-like immunoreactivity in the brain of the frog Rana ridibunda

The localization of atrial natriuretic factor (ANF)-like immunoreactivity in the central nervous system of the frog Rana ridibunda was examined by the indirect immunofluorescence technique, using an antiserum against synthetic ANF (Arg101-Tyr126). Immunoreactive cell bodies were principally found in the dorsal and medial pallium, the medial septal nucleus, the ventrolateral and anteroventral areas of the thalamus, the lateral forebrain bundle, the posterolateral thalamic nuclei, the preoptic nucleus, the dorsal infundibular nucleus, and the anteroventral tegmentum nucleus of the mesencephalon. Numerous cell bodies and a very dense fiber bundle were visualized in the interpeduncular nucleus. All the areas mentioned above contained a high density of immunoreactive fibers. In addition, the amygdala, the infundibular nucleus, the median eminence, and most of the areas of the mesencephalon contained a moderate number of ANF-positive nerve processes. In the frog pituitary, fibers and nerve terminals were found in the peripheral zone of the neural lobe. The intermediate and anterior lobes of the frog pituitary were totally devoid of ANF immunoreactivity. These results indicate that ANF-like material is widely distributed in the frog brain and that ANF may be involved in various brain functions including neuroendocrine regulations.     

Developmental study of neuropeptide Y-like immunoreactivity in the neurohypophysis and intermediate lobe of the rhesus monkey (Macaca mulatta)

Summary The purpose of this study was to examine the development and distribution of neuropeptide Y-immunoreactive fibers in the neurohypophysis of the rhesus monkey (Macaca mulatta) throughout life and the relationship of these fibers to the hypothalamo-hypophyseal portal vasculature. In rhesus monkeys, which varied in age from fetal life to 34 years, neuropeptide Y-immunoreactive fibers were present at all ages examined. In adult monkeys, varicose neuropeptide Y-labeled fibers were concentrated in the upper infundibular stem in association with capillary loops of the portal vasculature and the long portal vessels. Other fibers travelled down the infundibular stem and were distributed at the junction of the lower infundibular stem and infundibular process in the vicinity of the short portal vessels. In the infundibular process, neuropeptide Y-immunoreactive fibers were concentrated along the border of the intermediate lobe. Other stained fibers were sparsely distributed in the infundibular process and were often associated with small vessels. Neuropeptide Y-immunoreactivity was also located in a few fibers and cells of the intermediate lobe. Very few labeled fibers were seen in the fetal neurohypophysis, but their number increased gradually during the first postnatal year. At two years of age, a high density of stained fibers was observed, especially in the infundibular process. The number of axons in the infundibular process was lower at 12 years and continued to decline until 34 years of age. Neuropeptide Y may modulate hormone release at these sites and may also be released directly into vessels in the infundibular process. The close association of neuropeptide Y-labeled fibers with capillaries of the portal vasculature strongly suggests that neuropeptide Y is released into the portal blood of monkeys throughout life and may influence hormone secretion from the anterior pituitary gland.     

Effects of p-chloroamphetamine, a serotonin-depleting drug, on the median eminence and pituitary pars intermedia

p-Chloroamphetamine (PCA), an agent known to cause depletion of levels of brain serotonin in rodents, was administered to rats in three sequential injections (10mg/kg) to study effects on the hypothalamic median eminence and pituitary gland. One week following the initial sequence of injections of PCA, light and electron micrographs revealed degenerate fibers in the outer zone of the median eminence. Lower drug doses or single 10-mg/kg doses did not lead to morphologic changes. Neuronal processes located in the pituitary intermediate lobe appeared normal although there was a significant increase in the numbers of secretory granules contained within intermediate lobe cells drug-treated rats, as compared to controls. Fluorometric analysis of levels of catecholamine and indoleamine showed a decrease in serotonin in median eminence and pons-medulla, but no change in that of the pituitary. Levels of dopamine and norepinephrine remained unchanged after PCA treatment. The data suggest that fibers affected in the median eminence contain serotonin. Processes in the intermediate lobe may be resistant to the serotonin-lowering effects of PCA observed in brain tissue. In addition, PCA may directly affect granule release from pituitary cells, or may alternatively act on hypothalamic regions which affect the release of intermediate lobe cell hormones.     

Organization of tyrosine hydroxylase-immunoreactive neurons in the di-and mesencephalon of the American bullfrog (Rana catesbeiana) during metamorphosis

Summary We examined the immunocytochemical distribution of tyrosine hydroxylase, the rate-limiting enzyme in catecholamine synthesis, in the di-and mesencephalon of developing bullfrog tadpoles. Special attention was given to catecholaminergic innervation of the median eminence and pituitary. In premetamorphic tadpoles, tyrosine hydroxylase-immunoreactive neurons were visualized in the suprachiasmatic and infundibular hypothalamus, the ventral thalamus, and midbrain tegmentum by Taylor-Kollros stage V. The number of labeled neurons in all these areas increased as metamorphosis progressed. By mid-prometamorphosis, labeled neurons appeared in the preoptic recess organ as well as in the posterior thalamic nucleus. The majority of cells in the preoptic recess organ, as well as occasional neurons in the suprachiasmatic nucleus, exhibited labeled processes which projected through the ependymal lining of the preoptic recess to contact cerebrospinal fluid. The modified CSF-contacting neurons of the nucleus of the periventricular organ were devoid of specific staining. By late prometamorphosis, labeled fibers from the suprachiasmatic nucleus were observed projecting caudally to enter the hypothalamo-hypophysial-tract en route to innervating the median eminence and pituitary. Labeled fibers arising from the dorsal infundibular nucleus projected ventrolaterally to contribute to catecholaminergic innervation of the median eminence and pituitary. Immunoperoxidase staining of tyrosine hydroxylase-immunoreactive fibers and terminal arborizations in the median eminence were restricted to non-ependymal layers, while labeled fibers in the pituitary were observed in the pars intermedia and pars nervosa. Staining of tyrosine hydroxylase-immunoreactive fibers in the median eminence and pituitary was sparse or absent in premetamorphic tadpoles, but became increasingly more intense as metamorphosis progressed.     

Ultrastructural studies on the localization of neurohypophysial hormones and their carrier proteins.

Neurophysin, vasopressin and oxytocin were localized in different portions of the supraopticohypophysial tract (SHT) using the unlabeled antibody enzyme technique at the ultrastructural level. In vasopressin-positive supraoptic perikarya, vasopressin and neurophysin were present in all neurosecretory granules. Within the zona interna of the median eminence, vasopressin and neurophysin were present in two populations of axons, one with granules of 1300-1500 A and one with granules of 900-1300 A. Following exposure of thin sections of median eminence to antiserum to neurophysin, reaction products were present in granules and in the extragranular cytoplasm in the axons with larger granules; in all other cases reaction product was confined to the granules. Vasopressin-positive fibers were also presented in large numbers of the zona externa of the median eminence and many terminated on the pituitary primary portal plexus. A few oxytocin fibers were present on the portal capillaries in the infundibular stalk. In the posterior pituitary all axon profiles were neurophysin positive. Neurophysin was present as both a granular and cytoplasmic pool. Vasopressin-containing axons account for 90% of the neuronal elements in the posterior pituitary and oxytocin for the remaining 10%. Findings on the subcellular distribution of these peptides are related to current theories on transport and release of neurohormones.     

Distribution of immunoreactive somatostatin in the primate hypothalamus

Immunocytochemical methods were used to compare the localization of somatostatin (SRIF) in the human and rhesus monkey hypothalamus. The distribution of SRIF-containing cell bodies and fibers is similar in the two species. Perikarya are located predominantly in the periventricular region and to a lesser extent in the ventromedial nucleus. Fibers occur in dense clusters within the periventricular region, ventromedial nucleus, arcuate nucleus, median eminence, and pericommissural area of both species. Analysis of serial sections suggests that fibers originate from cells in the periventricular region, extend ventrally through the ventromedial and arcuate nuclei to terminate around the portal vessels of the infundibular stalk, and thereby participate in the regulation of anterior pituitary function. Somatostatinergic fibers are also found surrounding non-immunoreactive perikarya in the ventromedial nucleus and periventricular region of both primates. This arrangement may support somatostatin's postulated role as a neurotransmitter or neuromodulator. The strong similarity between the localization of hypothalamic SRIF in the human and rhesus monkey supports the use of the rhesus monkey as a model for the study of somatostatin as a neuroendocrine regulatory in the human.