Lateral evaginations from the third ventricle into the rat mediobasal hypothalamus: an amplification of the ventricular route.

In this work we report the existence of several evaginations extending out of the third ventricle within the mediobasal hypothalamus of the rat. In coronal sections, these evaginations appear as very narrow canaliculi integrating a canalicular system, which increases the contact surface between the ventricular lining and the nervous tissue. Consequently these evaginations enlarge the ventricular route for the transport of active principles present in the cerebrospinal fluid, such as (neuro)hormones and neurotransmitters. The mediobasal hypothalamus includes the arcuate nucleus and the median eminence (both involved in neuroendocrine mechanisms and in the regulation of pituitary function). A possible implication of our finding is that the neuroactive substance-containing ventricular cerebrospinal fluid may reach the intercellular spaces of the surrounding neuropil of the arcuate nucleus. According to literature these substances cross the ependyma of the lateral wall of the infundibular recess of the third ventricle. We suggest that such substances might also pass through the ependymal lining of the canalicular system, which displays the same ultrastructural characteristics as the rest of the ependyma of the lateral wall of the third ventricle. Therefore, the arcuate neurons may be influenced not only by synaptic inputs (afferent fibers) but also by non-synaptic diffusion neurotransmission (by means of neuroactive substances present in the cerebrospinal fluid).

Connections of the precommissural nucleus.

The connections of the precomissural nucleus (PRC) have been examined with anterograde and retrograde axonal tracing methods in the rat. Experiments with cholera toxin B subunit (CTb) indicate that the PRC shares a number of common afferent sources with the dorsolateral periaqueductal gray (PAG). Thus, we have shown that the nucleus receives substantial inputs from the prefrontal cortex, specific domains of the rostral part of the lateral septal nucleus, rostral zona incerta, perifornical region, anterior hypothalamic nucleus, ventromedial hypothalamic nucleus, dorsal premammillary nucleus, medial regions of the intermediate and deep layers of the superior colliculus, and cuneiform nucleus. Moreover, the PRC also receives inputs from several PAG regions and from neural sites involved in the control of attentive or motivational state, including the laterodorsal tegemental nucleus and the ventral tegmental area. The efferent projections of the PRC were analyzed by using the Phaseolus vulgaris-leucoagglutinin (PHA-L) method. Notably, the PRC presents a projection pattern that resembles in many ways the pattern described previously for the rostral dorsolateral PAG in addition to projections to a number of targets that also are innervated by neighboring pretectal nuclei, including the rostrodorsomedial part of the lateral dorsal thalamic nucleus, the ventral part of the lateral geniculate complex, the medial pretectal nucleus, the nucleus of the posterior commissure, and the ventrolateral part of the subcuneiform reticular nucleus. Overall, the results suggest that the PRC might be viewed as a rostral component of the PAG, and the possible functional significance of the nucleus is discussed in terms of its connections.

PSA-NCAM and B-50/GAP-43 are coexpressed by specific neuronal systems of the adult rat mediobasal hypothalamus that exhibit remarkable capacities for morphological plasticity.

The present study was designed to determine whether the mediobasal hypothalamus of adult rats contains neurons that continue to coexpress the highly polysialylated neural cell adhesion molecule (PSA-NCAM) and B-50/GAP-43, two proteins coexpressed by virtually all of the neurons of the fetal and neonatal rat central nervous system. Confocal laser scanning microscopy combined with double- or triple-fluorescence immunostaining was used to identify the hypothalamic neurons that express high levels of both PSA-NCAM and B-50/GAP-43 and to study the possible modifications of their morphological organization following a surgical lesion through the mediobasal hypothalamus. In intact animals, PSA-NCAM and B-50/GAP-43 were found to be colocalized within numerous fibers projecting throughout the external layer of the median eminence that were immunoreactive for either gamma-aminobutyric acid (GABA) or tyrosine hydroxylase (TH). Three to 30 days after a lesion through this region, numerous regenerating axonal sprouts, triple-immunostained for PSA-NCAM, B-50/GAP-43, and either GABA or TH, were detected along the ventricular surface of, and throughout the perivascular layer of, the median eminence. Surprisingly, high levels of PSA-NCAM and B-50/GAP-43 were also associated with numerous supraependymal neurons that exhibited long ramified processes and were immunoreactive for GABA but TH-negative. The use of the proliferation marker, 3H-thymidine, further indicated that the emergence of such supraependymal neurons after median eminence lesion was not related to the proliferation of preexisting quiescent cells. These data indicate that the mediobasal hypothalamus of the adult rat contains two neuronal systems, in which the continued coexpression of PSA-NCAM and B-50/GAP-43 is related to remarkable capacities for postlesional, morphological plasticity.

Efferent projections from the ovarian steroid receptor-containing area of the ventrolateral hypothalamus in female guinea pigs.

The ventrolateral hypothalamus (VLH) in female guinea pigs includes a subset of neurons which contain estrogen and progestin receptors, and which are implicated in the regulation of female sexual behavior by steroid hormones. However, little is known about where these neurons project, and consequently which other brain areas are involved in sexual behavior in female guinea pigs. The anterograde tracer Phaseolus vulgaris-Leucoagglutinin was used to label efferents from the ovarian steroid receptor-containing part of the VLH. To identify the correct placement of the tracer specifically within the group of neurons containing estrogen receptors, medial hypothalamic sections were also immunostained for estrogen receptors. Forebrain areas receiving dense projections from the ventrolateral hypothalamus included the bed nucleus of the stria terminalis, medial preoptic area, anterior hypothalamic area, anterior ventromedial hypothalamus, and caudal ventrolateral hypothalamus. The midbrain central gray was also heavily labeled. Moderate innervation was observed in the forebrain in the basolateral amygdala, medial preoptic nucleus, lateroanterior hypothalamic nucleus, dorsal hypothalamic areas, posterior hypothalamus, zona incerta, and in the midbrain interspersed among the central and lateral tegmental tracts. The major efferent pathways from the VLH appeared to travel rostrally through the mediobasal hypothalamus and preoptic area, and caudally via the medial thalamic nuclei and periventricular fiber system. These findings are similar to those of previous studies tracing the efferents from the ventromedial nucleus in rats and from the lateral hypothalamus in guinea pigs. Many of these areas that receive input from the steroid receptor rich area within the VLH are likely to be involved in the regulation of female sexual behavior.

Effects of ovariectomy on GnRH neuronal morphology in rhesus monkey (Macaca mulatta).

Gonadotropin releasing hormone (GnRH) neurons are typically simple, fusiform cells; however, over the course of prepubertal development increasing numbers take on a 'spiny' appearance. Following gonadectomy there is a decrease in the frequency of these spiny GnRH neurons. These observations which were made in the rat suggest that GnRH neurons are directly affected by the gonadal steroid milieu, though they do not themselves contain receptors for these steroidal hormones. In that there are important species differences in the hypothalamic-pituitary-gonadal axis between rats and primates, the present study was undertaken to determine whether a reduction in ovarian hormones would produce similar changes in the morphology of GnRH neurons in the monkey. A further aim was to determine whether such changes were localized to a specific brain region. Immunocytochemically defined GnRH neurons were compared in adult rhesus macaques which had been ovariectomized for 6 weeks to 2 years (n = 7) and intact, cycling animals (n = 8). Within the intact group, there were significantly more spiny GnRH neurons in the medial basal hypothalamus (MBH) than in the preoptic area (POA) (about 50% of the total in the MBH compared to 33% in the POA). Following ovariectomy the frequency of spiny cells in the MBH dropped to less than 30%, but was not significantly reduced in the POA. These results suggest that changes in systemic gonadal steroid levels result in changes in the morphology of GnRH neurons preferentially in the MBH, a region that is considered critical in the generation of GnRH pulsatile release in the monkey.

Lateral septal projections onto tubero-infundibular neurons in the hypothalamus of the guinea pig.

Efferent projections of the lateral septal nucleus (LS) to the preoptic area and the hypothalamus were identified in 20 female guinea pigs after iontophoretic injection of the anterograde axonal tracer Fluoro-Ruby. Tubero-infundibular (TI) neurons of the preoptic area and the hypothalamus were retrogradely labeled after intracardiac injection of Granular Blue or Fluoro-Gold. Magnocellular neurons of the supraoptic and paraventricular nuclei were also labeled. The double labeling procedure allowed an estimation of the extent of the direct relationship between LS efferents and TI neurons. Contacts between lateral septal fibers and TI cell bodies were mainly observed at the light-microscopical level in the preoptic area. A group of labeled fibers coursing along the third ventricle established sparse connections with hypothalamic periventricular TI neurons. A few appositions was observed in the infundibular (arcuate) nucleus, suggestive of a monosynaptic regulation of TI neurons by a septo-arcuate tract. Close association with labeled magnocellular neurons was also noted at the edge of the supraoptic and paraventricular nuclei. The sparse but direct connections between LS and TI neurons may be involved in the neuroendocrine functions of the LS.

[The development of a graft of embryonic mediobasal hypothalamus in the 3rd cerebral ventricle of the adult rat]. / Razvitie transplantata émbrional'nogo mediobazal'nogo gipotalamusa v 3-m zheludochke mozga vzrosloi krysy.

Development of the mediobasal hypothalamus from 15-day rat foetuses and 8-week human foetuses transplanted in the 3rd ventricle of the adult rat brain has been morphologically analyzed. The graft was shown to fill the ventral area of the 3rd ventricle and integrate with the host brain, as was especially distinct in the region of optic chiasma. The graft was abundantly vascularized and its vessels connected with the host brain vessels. The graft neurons were normally differentiating in situ. Some neurons migrated in the host brain. The graft neuropile ultrastructure was characterized by the abundance of synaptic contacts. Some graft neurons expressed dopaminergic phenotype by synthesizing tyrosine hydroxylase and DOPA-decarboxylase and displaying specific capture of 3H-dopamine. Dopaminergic axons of the neurons were spreading both within the graft and penetrating in the host tissue, especially in the region of optic chiasma and tracts. Unlike allotransplantation in rats, survival of xenotransplants of the human embryonic nervous tissue in the 3rd ventricle of the adult rat brain was possible only under the conditions of constant immunosuppression.

The ventricular system of the pigeon brain: a scanning electron microscope study.

The fine structural features and regional differences of the ependyma in adult pigeons have been investigated by scanning electron microscopy. Pigeons of either sex were fixed with buffered glutaraldehyde (3%) and formaldehyde (0.5%) by intravascular perfusion. The brain was dissected using section planes adequate to expose each part of the ventricular system. The specimens were then dehydrated, critical point dried and sputtered with gold. Depending upon the distribution of cilia, microvilli and single cilia, different areas were recognised in the 4 ventricles. The topographic locations of these areas were determined using the atlas of Karten & Hodos (1967). The medial surfaces of the 1st and 2nd lateral ventricles are more densely ciliated than the lateral surfaces. In the floor of the 4th ventricle the medial part is less ciliated than the lateral parts. The circumventricular organs (subseptal organ, organum vasculosum of the lamina terminalis, infundibulum, choroid plexus, subcommissural organ, area postrema) show very characteristic surfaces and are surrounded by a transitional zone with the nonspecialized ependyma. In contrast, in the paraventricular organ the transition to the nonspecialized ependyma is rather abrupt. The ependyma covering the trochlear nucleus appears densely ciliated, differing from that of the classic circumventricular organs. Finally, the existence of openings in the caudal medullary velum, which represent direct communications between the ventricles and the subarachnoid space, was demonstrated.

Distribution and morphology of immunoreactive gonadotropin-releasing hormone (GnRH) neurons in the basal forebrain of ponies.

Recent reports have indicated that analysis of changes in the staining characteristics of gonadotropin-releasing hormone (GnRH) neurons and characterization of morphological plasticity of the related structural framework may help to elucidate the physiological mechanisms involved in neuroendocrine control of mammalian reproduction. Whether comparative studies will facilitate this process or simply elucidate species-specific mechanisms is not yet clear. The present study was performed in order to begin analysis of GnRH neurons in a seasonally breeding species that exhibits an unusually long ovulatory luteinizing hormone (LH) surge. To this end, light microscopy and image analysis were used to characterize distribution and morphology of GnRH neurons in 15 adult male and female ponies. Samples were collected in the middle of the normal ovulatory season. Unipolar, bipolar, and multipolar GnRH neurons were organized in a loosely defined continuum that extended from the medial septum to tuberoinfundibular areas in the medical basal hypothalamus (MBH). Most cells were bipolar, and the majority of neurons were located in the MBH. Fiber projections to the median eminence included presumptive pathways similar to those previously described in other species. Image analysis of cell size indicated that cells in the MBH were larger than those in preoptic areas and GnRH neurons in both of these locations were larger than neurons in rostral areas of the medial septum. Results from this experiment suggest that the large population of MBH GnRH neurons in the equine species is likely to be of primary importance to reproductive function, whereas cells in other areas are fewer and smaller. Further work is needed to characterize morphological characteristics that may be related to physiological fluctuations in reproductive function of the equine species.

Modulation of feeding by hypothalamic paraventricular nucleus alpha 1- and alpha 2-adrenergic receptors.

Noradrenergic receptor populations within the paraventricular hypothalamus (PVN) modulate feeding. Satiated rats exhibit enhanced feeding subsequent to activation of alpha 2-adrenergic receptors within the PVN induced by exogenous infusion of either norepinephrine (NE) or clonidine (CLON). The feeding-stimulatory effect of alpha 2-adrenergic agents presumably reflects an inhibitory action on receptors located on medial hypothalamic "satiety" cells. Adrenergic receptors of the alpha 1-subclass have been identified within the PVN which are excitatory and which may function to suppress food intake. Microinjection into rat PVN of various alpha 1-adrenergic agonists including cirazoline, methoxamine, phenylpropanolamine and phenylephrine suppress feeding; an effect that is reversed by pretreatment with alpha 1-adrenergic receptor antagonists. The present review argues that alpha 1- and alpha 2-adrenoceptors within brain and specifically within the PVN are organized in an antagonistic fashion and that the effects of various adrenergic agonists on feeding may reflect the degree to which these agonists act at alpha 1- and alpha 2-adrenoceptors as well the relative balance of these receptors and their activity within the PVN.

Neuropeptide-Y innervation of beta-endorphin-containing cells in the rat mediobasal hypothalamus: a light and electron microscopic double immunostaining analysis.

Central administration of neuropeptide-Y (NPY) inhibits pituitary LH release in ovariectomized rats and stimulates LH release in intact and ovariectomized rats pretreated with ovarian steroids. Although the precise neural mechanism of this dual effect of NPY is not known, experimental evidence suggests an underlying interaction between hypothalamic NPY and the inhibitory beta-endorphin (beta END) systems in the neuroendocrine regulation of pituitary LH release in the rat. The present study was undertaken to examine the morphological basis of the interaction between these two peptidergic systems in the hypothalamus. Sections of the mediobasal hypothalamus of colchicine-pretreated female rats were double immunostained for NPY and beta END and examined by light and electron microscopy. The light brown diaminobenzidine reaction was used to visualize beta END cells, while NPY neurons were labeled with a dark blue nickel ammonium sulfate-intensified diaminobenzidine reaction. Under the light microscope, a dense network of NPY-immunoreactive axons and axon terminals was observed in close apposition with beta END-immunoreactive neurons throughout the medial basal hypothalamus. Electron microscopic examination revealed that NPY-immunoreactive boutons formed axosomatic and axo-dendritic synaptic connections with beta END cells. A majority of these synaptic membrane specializations appeared asymmetrical [corrected]. In light of the previous evidence of excitatory and inhibitory effects on LH release and the existence of direct synaptic connections between NPY and LHRH neurons in the hypothalamus, the current results imply that the dual effects of NPY on LH secretion may involve modulation of LHRH secretion, both by the direct route and indirectly through the hypothalamic beta END system.

GABAergic and catecholaminergic innervation of mediobasal hypothalamic beta-endorphin cells projecting to the medial preoptic area.

In the absence of cellular estrogen receptors or proven direct estrogen action in the rat, it is assumed that estrogen indirectly regulates the secretory activity of the preoptic area luteinizing hormone-releasing hormone-producing cells. We have previously shown that pro-opiomelanocortin neurons in the arcuate nucleus of the rat send axons rostrally to connect with luteinizing hormone-releasing hormone neurons of the preoptic area. An experiment combining retrograde tracing and double-immunostaining was used to test the hypothesis that rat GABAergic and/or catecholaminergic neurons can influence luteinizing hormone-releasing hormone-producing cells via mediobasal hypothalamic beta-endorphin neurons. The retrograde tracer horseradish peroxidase was injected into the medial preoptic area; two days later, arcuate nucleus Vibratome sections were double-immunostained for beta-endorphin and glutamate decarboxylase or tyrosine hydroxylase. Light and electron microscopic analysis of these triple-labeled sections demonstrated that a population of beta-endorphin-immunoreactive neurons concentrated in the ventromedial arcuate nucleus contain retrogradely transported horseradish peroxidase granules and form synaptic contacts with glutamate decarboxylase- and tyrosine hydroxylase-immunoreactive axon terminals. The present data suggest that arcuate nucleus GABA and catecholamine fibers may influence luteinizing hormone-releasing hormone-containing neurons via projective pro-opiomelanocortin cells.

Presence of calbindin and lack of parvalbumin in progesterone receptor-containing neurons of the monkey mediobasal hypothalamus.

All of the progesterone receptor-containing cells of the monkey hypothalamus are GABAergic. The aim of this study was to further characterize these GABAergic progesterone receptor-containing neurons based on their calbindin or parvalbumin content. These calcium-binding proteins are characteristic markers of different populations of GABAergic neurons in the central nervous system. Double-immunolabeling for progesterone receptor and either calbindin or parvalbumin was performed on hypothalamic Vibratome sections of estrogen primed African green monkeys (Cercopithecus aethiops). Progesterone receptor-containing calbindin-immunoreactive neurons were observed in the ventromedial and periventricular areas of the hypothalamus. Forty-one per cent of the progesterone receptor-containing cells in this area were calbindin immunopositive. No double-immunolabeled neurons could be detected in the infundibular (arcuate) nucleus. In tissue double-immunolabeled for progesterone receptor and parvalbumin, none of the progesterone receptor-containing neurons exhibited immunoreactivity for parvalbumin. Electron microscopic double-immunostaining for progesterone receptor and calbindin confirmed the light microscopic results. Furthermore, a large number of asymmetric synaptic contacts were observed on the calbindin-immunoreactive neurons. These observations demonstrate that progesterone receptor-containing cells in the monkey mediobasal hypothalamus consist of at least two different types of GABA neurons, and indicate that progesterone receptor-containing calbindin cells may be postsynaptic targets of excitatory fibers.

[Morphologic substrates of the ventricular route of secretion and transport of substances in the tubero-infundibular region of the hypothalamus. Ultrastructural study]. / Sustratos morfológicos de la vía ventricular de secreción y transporte de sustancias en la región tuberoinfundibular del hipotálamo. Estudio ultraestructural.

Ultrastructural studies of the ependyma of the tuberoinfundibular region of the rat hypothalamus have revealed the existence of intraventricular axonal endings and of cytoplasmic blebs and bulbs that project from the apical surface of the ependymal cells to the ventricular lumen. All these structures account for the processes of ependymal apocrine secretion and the neuroventriculocrinia, and hence the release of biologically active substances into the cerebrospinal fluid (CSF). These substances contained in the CSF must act on the nervous nuclei of the tuberoinfundibular region, such as the arcuate nucleus, which is very important in the neuroendocrine regulation of the anterior pituitary gland. Dilated intercellular spaces among neighbouring ependymocytes of this region, small intraependymal cisternae and, in particular, a lateral prolongation of the infundibular recess, which courses through the nervous tissue between the arcuate nucleus and the median eminence from the vertex of the lateral angle of the infundibular recess, may be the route followed by the CSF from the third ventricle to the tissue compartment of the tuberoinfundibular region. Also studied are the cisternae of the region and the relationships of these with the lateral prolongation of the infundibular recess. Some of these cisternae may be filled by the CSF through the prolongation. In this way, the tissue compartment of CSF would be enlarged, and hence the ventricular route for the secretion and transport of biologically active substances would be potentiated.

Effects of gonadal steroids on the ultrastructure of GnRH neurons in the rhesus monkey: synaptic input and glial apposition.

The secretion of the gonadotropins is modulated by the gonadal steroids, but the means by which these effects are mediated are not well understood. The present anatomical study was undertaken to investigate the possibility that the GnRH system responds to alterations in the gonadal steroid environment with reversible changes in synaptic input and glial wrapping such as have been observed in other neuroendocrine systems. The ultrastructure of GnRH neurons was studied in the preoptic area and medial basal hypothalamus of rhesus monkeys in various steroid conditions including five intact cycling, four long-term ovariectomized animals, two long-term ovariectomized animals with steroid replacement (LtOVX+), and two animals replaced with steroid at the time of ovariectomy (StOVX+). Electron micrographic montages of GnRH neuronal profiles were analyzed using computerized morphometrics, and the percentages of the length of perikaryal membrane immediately apposed by glial processes and that with postsynaptic modification were calculated. Ovariectomy resulted in a significant increase in the apposition of glial processes to GnRH perikaryal membranes and a significant decrease in their innervation in both brain regions. There was also a higher incidence of GnRH neurons with immunostaining confined to secretory granules and a decrease in the volume of nucleoli, both of which could be interpreted as indications that GnRH peptide synthesis was reduced in ovariectomized animals. After an ovarian steroid replacement regimen which mimicked two menstrual cycles, the innervation of GnRH neurons was increased and the glial ensheathment was partially reduced. This was true for both the LtOVX+ and StOVX+ steroid-replacement groups. GnRH neurons in the medial basal hypothalamus received more synaptic input than did those in the preoptic area, regardless of the steroid condition of the animal. The degree of glial ensheathment of GnRH neurons in the preoptic area became significantly greater than that in the medial basal hypothalamus after ovariectomy. These observations suggest there may be differences in the role of GnRH neurons in these two brain regions. These immunocytochemical ultrastructural studies provide strong evidence that alterations in the gonadal steroid milieu can produce morphological changes in the GnRH neuron and its immediate environment in the primate.

Estrogen-enhanced neurite growth: evidence for a selective induction of Tau and stable microtubules.

Estrogen stimulates the neurite outgrowth response of medial basal hypothalamic neurons maintained in culture. We show here that one effect of estrogen is to promote an increase in tau, but not in tubulin, microtubule-associated protein 1a (MAP-1a), or MAP-2 protein levels. This response precedes and accompanies an increase in stable microtubules and in neurite length. Taken collectively, our data suggest that estrogen-enhanced neurite growth is mediated by a selective induction of microtubule-stabilizing factors, namely, the tau proteins.

Estradiol increases the dendritic length of ventromedial hypothalamic neurons in female Syrian hamsters.

We examined the effects of ovarian hormones on dendritic architecture of neurons in the ventromedial nucleus of the hypothalamus in female Syrian hamsters. Treatment with 10 micrograms of estradiol benzoate for two days, or estradiol benzoate for two days followed by an injection of 500 micrograms of progesterone, increased the total dendritic length of ventromedial nucleus neurons by almost 50% compared with neurons from the ventromedial nucleus of ovariectomized, oil-treated females. Neurons in a control region, the dorsomedial nucleus of the hypothalamus, were unaffected by these hormone treatments. These results demonstrate that steroids can induce changes in dendritic structure within 48 hr, suggesting that such morphological reconfiguration of hypothalamic neurons may underlie variations in behavior associated with the female's 4-day estrous cycle.

Gonadal steroids modify dendritic spine density in ventromedial hypothalamic neurons: a Golgi study in the adult rat.

Neurons in the adult rat ventromedial hypothalamic nucleus (VMN, 4-6 neurons per brain; 3-7 brains per group) were studied under various hormonal conditions using the single-section Golgi impregnation technique. Intact rats of both sexes and ovariectomized females treated with oil, estrogen or estrogen and progesterone were used. Golgi-impregnated neurons in the VMN were analyzed to determine possible differences in cell body size, number of primary dendrites, number of dendritic branch-points and spine density. The only significant differences found were in spine density. In the VMN of ovariectomized rats given oil, there were significantly fewer spines on primary dendrites than in either estrogen-treated, estrogen plus progesterone-treated or intact female rats. There were no differences between intact male and female rats observed in any parameter. In addition, when spine density of VMN neurons was assessed throughout the estrous cycle, it was determined that spine density was significantly lower at diestrus than proestrus.

Ultrastructural characterization of prolactin-like immunoreactivity in rat medial basal hypothalamus.

Prolactin-like immunoreactivity has been reported in the medial basal hypothalamus at the light microscopic level, in hypophysectomized rats. Here, with preembedding immunocytochemistry at the electron microscopic level, we have observed prolactin-immunoreactive neurons and synapses in the hypothalamus. Reaction product was discovered in medial basal hypothalamic neurons, which had typical large nucleoli and received axosomatic synapses. In the cytoplasm, reaction product was distinctly granular. Immunoreactive neurons were usually surrounded by nonreactive cells. Reaction product was also seen in dendrites, some of which had spines. Some axons in the hypothalamus contained reaction product, usually surrounded by nonreactive axons, and immunopositive synapses were detected both in the hypothalamus and in the midbrain. In a small number of cases immunoreactive axons could be seen synapsing on immunoreactive dendrites.

The bifunctional role of pro-opiomelanocortin derivatives in the mediobasal hypothalamus of the rat.

In the present study, a polyclonal antibody against pro-opiomelanocortin derivatives was characterized biochemically. Its immunoreactivity with structures of the arcuate nucleus and the median eminence was investigated by means of the immunogold method and compared with its reaction on adenohypophyseal cells with and without pre-adsorption with pro-opiomelanocortin derivatives. The antiserum detects ACTH and its fragments, in particular alpha-MSH, and beta-endorphin. In the adenohypophysis gold particles are exclusively located on small secretory granules situated in the periphery of branched cells. In the perikarya of the arcuate nucleus gold particles are observed on terminal vesicles abutting from the cis-face of the Golgi apparatus, on granules in its direct vicinity and on small dense core vesicles preferentially located in the cell periphery. Immunoreactive gold-labeled fiber profiles are found in a sub- or intra-ependymal position as well as in the nuclear neuropil proper. Here axodendritic and axosomatic synapses are observed. In both situations the gold particles are mostly restricted to the small dense core vesicles and do not decorate the synaptic vesicles. In the median eminence gold labeled fibers are detected in all layers. The labeled fibers can be closely apposed to tanycytic processes, without, however, forming special contact differentiations. In direction to the perivascular layer of the external zone the labeled profiles are more frequently arranged in groups intermingled with unlabeled fibers. The axons decorated with gold particles can be freely exposed to the perivascular space or are found as single processes in close vicinity to the capillary wall. Subsequent to preincubation of the native antiserum with ACTH1-39 and ACTH18-39 (= CLIP) neither adenohypophyseal cells nor perikarya and fibers in the arcuate nucleus nor axons in the median eminence are decorated with gold particles. Preincubation of the native antiserum with alpha-MSH or beta-endorphin does not change the immunoreaction with the small, peripherally situated granules in the branched adenohypophyseal cells. In neurons of the arcuate nucleus and in fibers of the median eminence, however, the immunoreaction is completely extinguished when the antibody is pre-incubated with alpha-MSH, whereas subsequent to preincubation with beta-endorphin only the amounts of labeled structures are reduced.(ABSTRACT TRUNCATED AT 400 WORDS)