Definitive evidence for the existence of morphological plasticity in the external zone of the median eminence during the rat estrous cycle: implication of neuro-glio-endothelial interactions in gonadotropin-releasing hormone release

Despite intense investigation, the demonstration of morphological plasticity in the external zone of the median eminence concerning the gonadotropin-releasing hormone system has never been reported. In this study, we investigate whether dynamic transformations of the gonadotropin-releasing hormone nerve terminals and/or tanycytes in the external zone of the median eminence of the hypothalamus occurred during the rat estrous cycle, by following individual gonadotropin-releasing hormone-immunoreactive nerve terminals on serial ultrathin sections observed by electron microscopy. Female rats were killed at 16.00 diestrus II (n = 3), i.e. when estrogen levels are basal and gonadotropin-releasing hormone release is low, and at 16.00 proestrus (n = 4), i.e. when estrogen levels peak and the preovulatory gonadotropin-releasing hormone surge occurs. Our results show that, in the median eminence obtained from proestrus rats, 12+/-2% of the gonadotropin-releasing hormone nerve terminals were observed to make physical contact with the parenchymatous basal lamina, i.e. the pericapillary space. In the median eminence obtained from diestrus II rats, no contacts were observed. On proestrus, numerous physical contacts between gonadotropin-releasing hormone nerve terminals and the basal lamina occurred by evagination of the basal lamina and/or by emerging processes from gonadotropin-releasing hormone nerve terminals. The quantification of the evagination of the basal lamina revealed that the basal lamina was at least twofold more tortuous in appearance during proestrus. These results demonstrate for the first time the existence of dynamic plastic changes in the external zone of the median eminence, allowing gonadotropin-releasing hormone nerve terminals to contact the pericapillary space on the day of proestrus, thus facilitating the release of the neurohormone into the pituitary portal blood.     

Topographic relations between tyrosine hydroxylase- and luteinizing hormone-releasing hormone-immunoreactive fibers in the median eminence of adult rats

Employing electron microscopic double immunolabeling, we determined a close apposition of tyrosine hydroxylase (TH) and luteinizing hormone-releasing hormone (LHRH) nerve fibers in the rat median eminence (ME). These axo-axonic contacts occurred frequently in the internal and palisade zones, i.e. at the level of the fiber preterminals. In the superficial area of the ME, major TH fibers abutted on the basal lamina and some were projected into the pericapillary space of the portal vessels. Conversely, LHRH fibers were arrested by the endfeet of tanycytes in reaching the basal lamina.     

The hypothalamic magnocellular neurosecretory system of the guinea pig. II. Immunohistochemical localization of neurophysin and vasopressin in the fetus

The development of the hypothalamic magnocellular neurosecretory system of the fetal guinea pig was examined by immunohistochemistry. Neurophysin was first observed in the supraoptic nucleus (SON), median eminence (ME) and posterior pituitary (PP) on day 40 of gestation. It was not regularly present in the paraventricular nucleus (PVN) until day 47. Vasopressin was first observed in the SON, ME and PP on day 45. In the median eminence immunoreactive deposits indicative of both peptides were observed in both the fibers of the hypothalamo-hypophysial tract (H-HT) in the presumptive zona interna as well as in axons projecting to the developing primary portal plexus.     

Immunohistochemical colocalization of delta sleep-inducing peptide and luteinizing hormone-releasing hormone in rabbit brain neurons

The anatomical distributions of luteinizing hormone-releasing hormone and delta sleep-inducing peptide immunoreactivity in the rabbit brain were studied by indirect immunofluorescence technique. The comparison of adjacent serial sections, one being immunolabeled with an antiserum to luteinizing hormone-releasing hormone, the other with an antiserum to delta sleep-inducing peptide, showed that the respective distribution patterns of immunoreactivity exhibited a remarkable overlap through the basal forebrain and hypothalamic regions. A sequential double-immunolabelling (elution-restaining method) clearly indicated that all the luteinizing hormone-releasing hormone-immunoreactive cell bodies displayed delta sleep-inducing peptide immunoreactivity. These cell bodies were sparse and mainly located throughout the septal-preoptico-suprachiasmatic region and the ventrolateral hypothalamus. The colocalization of luteinizing hormone-releasing hormone and delta sleep-inducing peptide immunoreactivity was also observed in many fibres supplying all these brain regions and terminal areas such as the organum vasculosum of the lamina terminalis, the subfornical organ, the median eminence and the pituitary stalk. These neuroanatomical findings are suggestive of interaction between delta sleep-inducing peptide and luteinizing hormone-releasing hormone in various brain areas including some circumventricular organs.     

Axonal projections from the hypothalamus to the median eminence in rats during ontogenesis: DiI tracing study

This study has determined the ontogenetic schedule of the arrival of the axons from the hypothalamus and the diagonal band in the median eminence in rats by using the fluorescent lipophilic carbocyanine dye, 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine perchlorate (DiI) as a retrograde tracer. After fixation of the brain, the crystals of the dye were implanted in the median eminence on the 13th, 14th, 15th, 16th,17th, 20th embryonic days, and on the 2nd postnatal day. This was followed by fluorescent staining of the neuronal cell bodies in the hypothalamus. According to our data, the axons of rare hypothalamic neurons first reached the primordium of the median eminence on the 14th embryonic day. For two subsequent days, the number of neurons projecting the axons to the median eminence appeared to increase considerably.They werewidely distributed through the hypothalamus and in the ventromedial region of the more rostral forebrain. Till the 20th embryonic day, the majority of the fluorescent neurons were concentrated mainly in the paraventricular nucleus (dorsal and medial parts) and the arcuate nucleus, and to a lesser extent in the medial preoptic nucleus, the supraoptic nucleus, the diagonal band, and the retrochiasmatic nucleus. In neonates, DiI-labelled neurons appeared additionally in the accessory dorsolateral nucleus,medial preoptic area lateral to the diagonal band, anterior hypothalamic area, and in the anterior periventricular nucleus. Thus, the axons of differentiating neurons arrive in the median eminence from the 14th embryonic day till the neonatal period, providing the pathway for the neurohormone transfer to the hypophysial portal circulation. Accepted: 12 March 2001     

Access of luteinizing hormone-releasing hormone neurons to the vasculature in the rat

Luteinizing hormone-releasing hormone is secreted into the hypophysial portal vasculature through which it controls the release of the gonadotropins. The peptide also acts as a neurotransmitter in various loci within the central nervous system. It is not known whether these roles are performed by separate populations of luteinizing hormone-releasing hormone neurons. Some recent tracing experiments suggest that this is the case (Silverman et al., J. Neurosci. 7, 2312, 1987; Jennes and Stumpf, Neuroscience 18, 403, 1986). One aspect of this question was addressed in the current study by intraperitoneal introduction of Fluoro-Gold (a retrograde tracer) into male and female rats under various age and hormonal conditions. Brain sections from the anterior olfactory nuclei to the median eminence were treated for the immunocytochemical demonstration of luteinizing hormone-releasing hormone. In all cases, regardless of the age, sex or hormonal condition of the animal, the Fluoro-Gold tracer was found in more than 90% of the luteinizing hormone-releasing hormone neurons. We conclude that virtually all luteinizing hormone-releasing hormone neurons in the rat secrete outside the blood-brain barrier, including those which project to sites within the central nervous system.     

Distribution and colocalization of delta sleep-inducing peptide and luteinizing hormone-releasing hormone in the aged human brain: an immunohistochemical study

The distributions of delta sleep-inducing peptide (DSIP)- and luteinizing hormone-releasing hormone (LHRH)-immunoreactive neurons were investigated in the human brain with special emphasis on the basal forebrain (from the septum to the hypothalamus), using indirect immunofluorescence. With a modified elution technique, sequential stainings on the same section showed that DSIP- and LHRH-immunoreactivities were often colocalized. Small numbers of LHRH/DSIP-immunoreactive cells were essentially detected in the diagonal band of Broca, the medial septum and the ventral hypothalamus. The richest areas displaying fibres and terminal-like structures were the preoptic area, the ventromedial and ventrolateral hypothalamic areas, the periventricular region and certain circumventricular organs (i.e. median eminence, vascular organ of the lamina terminalis). Few isolated fibres were observed in the subfornical organ. The topographical relationships between DSIP- and LHRH-immunoreactivities in the neurosecretory systems suggest that DSIP may play a role as important as that of LHRH.     

Gonadotrophin-releasing activity of the median eminence at different ages

1. Intrapituitary infusions of extracts made from the median eminence of various species at different ages were found to excite secretion of gonadotrophic hormone as shown by consequent ovulation, presumably due to their content of luteinizing hormone-releasing factor (LRF).2. Extracts from very immature animals (e.g. new-born rabbits and rats; 1-2-month-old calves) were found to be highly active with regard to LRF content.3. Extracts from foetal rabbits at 20 days of gestation showed no evidence of containing LRF.4. It is suggested that (a) LRF may have a trophic action upon the anterior pituitary gland during the pre-puberal phase of life, and that (b) the first formation of LRF coincides with the development of the primary plexus of the pituitary portal system.     

Subpopulations of corticotropin-releasing hormone neurosecretory cells distinguished by presence or absence of vasopressin: confirmation with multiple corticotropin-releasing hormone antisera

Parvocellular corticotropin-releasing hormone neurosecretory cells in the hypothalamic paraventricular nucleus project axons to the portal capillary plexus in the external zone of the median eminence. Immunocytochemical studies have identified two approximately equal subpopulations of these corticotropin-releasing hormone neurons in normal rats, distinguished by the presence or absence of co-existent vasopressin, and different responses to stress. However, it was recently proposed that the vasopressin deficient cells do not contain corticotropin-releasing hormone, but have been misidentified due to cross-reactivity of the corticotropin-releasing hormone antiserum to peptide histidine-isoleucineamide. It is shown here that the same set of corticotropin-releasing hormone neurons (including both vasopressin expressing and vasopressin deficient subtypes) was labeled with multiple corticotropin-releasing hormone antisera. These included two antisera that did not cross-react with peptide histidine-isoleucineamide: one against ovine corticotropin-releasing hormone, and one rat corticotropin-releasing hormone antiserum absorbed with peptide histidine-isoleucineamide. The results provide further support for the hypothesis of functionally distinct compartments of the corticotropin-releasing hormone neurosecretory system that can modulate the ratio of vasopressin to corticotropin-releasing hormone in portal blood.     

The hypothalamic magnocellular neurosecretory system of the guinea pig. I. Immunohistochemical localization of neurophysin in the adult

With the use of the unlabeled antibody enzyme technique and antiserum against bovine neurophysin I, neurophysin was localized in the hypothalamic magnocellular neurosecretory system of the adult guinea pig. Immunoreactive deposits were found in the perikarya of the supraoptic and paraventricular nuclei, their fiber projections and terminals in the posterior pituitary. No parvicellular neurophysin-positive components were observed. In the median eminence neurophysin was seen in the zona interna where axons of the supraoptico-hypophysial tract pass on their way to the neural lobe. The peptide was also present in axons projecting into zona externa which terminate on the primary portal plexus.     

Ontogenetic development of TRH-like immunoreactive nerve terminals in the median eminence of the rat

Ontogenetic development of TRH-like immunoreactive nerve terminals in the median eminence of the rat was studied immunocytochemically. By light microscopy, TRH-like immunoreactivities were first detected on the 1st day after birth in the external layer of the median eminence. By electron microscopy, TRH-like immunoreactive nerve fibers and terminals were visible on the 0.5th day after birth. The nerve terminals were first found in direct contact with the perivascular basal lamina of the portal vessel on the 2nd day. TRH-like immunoreactivities were only localized on dense granular vesicles about 105 nm in diameter in the axoplasm throughout the developmental stages. The immunoreactive nerve fibers with TRH-like immunoreactive granular vesicles gradually increased in number with development. The physiological significance of TRH as a hormone is discussed in relation to the presence of TRH-like immunoreactive nerve terminals in the median eminence of the developing rat.     

Demonstration of serotoninergic axons terminating on luteinizing hormone-releasing hormone neurons in the preoptic area of the rat using a combination of immunocytochemistry and high resolution autoradiography

The synaptic relationship between serotoninergic terminals and luteinizing hormone-releasing hormone-containing neurons was investigated in the medial preoptic area using a combined technique. Axon terminals selectively taking up 5-[3H]hydroxytryptamine were labelled autoradiographically and luteinizing hormone-releasing hormone-containing neuronal elements were identified by means of immunocytochemistry. Synaptic contacts were observed between tritiated 5-hydroxytryptamine-labelled boutons and luteinizing hormone-releasing hormone-immunoreactive dendrites. About 5% of the boutons which formed synapses with luteinizing hormone-releasing hormone-immunoreactive dendrites were found to be labelled by the tritiated indolamine. Luteinizing hormone-releasing hormone-immunoreactive axon terminals occurred as presynaptic elements in contact with unidentified dendritic spines, shafts or perikarya. These observations provide morphological basis for the idea that 5-hydroxytryptamine-containing neurons can act directly on luteinizing hormone-releasing hormone release. Further, they support the assumption that luteinizing hormone-releasing hormone is not only a neurohormone but may also function as a neurotransmitter or neuromodulator.     

Immunohistochemical localization of luteinizing hormone-releasing hormone (LHRH) in the hypothalamus of adult female hamsters treated neonatally with monosodium glutamate or hypertonic saline

The immunohistochemical localization of luteinizing hormone-releasing hormone (LHRH) was studied in paraffin and vibratome-sectioned tissue from adult female hamsters that were treated neonatally with monosodium glutamate (MSG) or hypertonic saline. There appeared to be a reduction in LHRH-positive fibers in the median eminence of animals with an MSG-induced lesion of the arcuate nucleus in paraffin-embedded tissue. However, when unembedded tissue was cut on a vibratome, the distribution of LHRH-positive fibers and perikarya was similar in both groups of animals. Fibers were seen coursing through the periventricular area and lateral hypo-thalamus to the median eminence. In addition, LHRH-positive fibers were seen in the organum vasculosum of the lamina terminalis, subfornical organ, septal and preoptic areas, fasciculus retroflexus, habenular complex, and several regions in the basal forebrain. Animals that were pretreated with colchicine had LHRH-positive perikarya in the medial habenular nucleus, diagonal band of Broca, and the medial olfactory tract.     

Localization of luteinizing hormone-releasing hormone in the mammalian hypothalamus

Utilizing immunohistochemistry with rabbit antiserum to synthetic luteinizing hormone-releasing hormone (LRH), LRH was localized in the peripheral region of the median eminence in the mouse and rat, and more generally in the median eminence of the guinea pig.     

Implantation of fetal preoptic area into the lateral ventricle of adult hypogonadal mutant mice: the pattern of gonadotropin-releasing hormone axonal outgrowth into the host brain

Transplantation of fetal preoptic area tissue containing gonadotropin-releasing hormone neurons into the third ventricle of male hypogonadal mice resulted in an elevation of pituitary gonadotropin levels and correction of hypogonadism. This reversal of the neuroendocrine deficit was correlated with innervation of the median eminence by gonadotropin-releasing hormone axons. The specificity of fiber outgrowth suggested that local neuromodulatory factors might guide these axons to the nearby median eminence. To test this hypothesis, 14 adult hypogonadal males received unilateral fetal preoptic area grafts into the lateral ventricle, a site distant from the median eminence. After four months, healthy grafts containing numerous gonadotropin-releasing hormone neurons were seen in 9 hosts. However, none of these grafts corrected the hypogonadism of the host and there was no gonadotropin-releasing hormone innervation of the median eminence in any of these animals, thus demonstrating that the presence of gonadotropin-releasing hormone neurons in the ventricular space is itself not sufficient to stimulate the pituitary-gonadal axis. Instead, gonadotropin-releasing hormone axons coursed in the host fimbria, fornix, corpus callosum, and stria terminalis. These fibers could be traced into the anterior hippocampal area, medial and lateral septum, and the anterior hypothalamus. The distribution of these fibers included a number of regions which receive gonadotropin-releasing hormone fiber input in the normal mouse. These findings show that gonadotropin-releasing hormone neurons transplanted into the lateral ventricle can survive and extend processes into the host brain, often projecting to sites of normal gonadotropin-releasing hormone innervation. Their success in contacting these sites suggests that gonadotropin-releasing hormone fiber outgrowth may be influenced by regionally specified trophic and/or guidance factors.     

Implantation of normal fetal preoptic area into hypogonadal mutant mice: temporal relationships of the growth of gonadotropin-releasing hormone neurons and the development of the pituitary/testicular axis

Central nervous system tissue which included the preoptic area (an area rich in gonadotropin-releasing hormone neurons) was taken from normal 17-day fetal mice and transplanted into the infundibular recess of the third ventricle of the hypothalamus of 90-day male mutant hypogonadal mouse hosts that are unable to synthesize the neurohormone, gonadotropin-releasing hormone. The growth and development of gonadotropin-releasing hormone neurons and fibers in the donor and host tissue as well as recovery of the pituitary-testicular axis were followed from 10 to 120 days post-implantation. Testicular growth was evident in 94% of the hypogonadal animals within 30 days post-implantation, continued for 90 days but showed no further increase during the remainder of the experiment. Increases in seminal vesicle weight, an index of testosterone secretion, were measurable at 30 days and continued through to the end of the experiment. Pituitary concentrations of gonadotropins were doubled at 30 days over that seen in the control mutant mouse and were maintained thereafter at normal or supranormal concentrations. In contrast plasma levels of gonadotropins, although above baseline at 30 days, never reached normal circulating levels. Nevertheless, it appeared that the concentration of luteinizing hormone achieved was sufficient to initiate and maintain testicular growth and testosterone secretion for the entire duration of the experiment. Immunocytochemical analysis of brain tissue was used to determine the presence and numbers of gonadotropin-releasing hormone neurons in the transplant and the distribution of their fibers in the donor and host tissue. The numbers of immunoreactive gonadotropin-releasing hormone neurons present at the time of sacrifice ranged from 3 to 140. Fiber outgrowth from the donor cells into the host was noted as early as 10 days post-implantation and the density of outgrowth continued to increase over the course of the experiment. Positive fibers tended to accumulate over the tuberoinfundibular sulci as they do in normal animals. In those instances where the transplant was placed a long distance from the median eminence, the gonadotropin-releasing hormone axons grew on the internal surface of the third ventricle until they reached these specific exit zones. These studies indicate that in the mutant hypogonadal mouse, central nervous system transplants from normal fetal mice can maintain the function of the pituitary-gonadal axis for periods of up to 120 days post-implantation. Outgrowth of the neurosecretory fibers begins very soon after implantation and the axons tend to follow pathways seen in normal tissue.(ABSTRACT TRUNCATED AT 400 WORDS)     

Morphological adaptability at neurosecretory axonal endings on the neurovascular contact zone of the rat neurohypophysis

To compare the effects of a variety of acute and chronic stimuli that bring about or terminate hormone release the ultrastructure of nerve terminal contact at the basal lamina of the neurohypophysial neurovascular contact zone was examined quantitatively in young adult rats of the following treatment groups: untreated virgin females, untreated male rats, prepartum (day 21 of gestation), postpartum (on the day of parturition), lactating (14 days of suckling), mothers 10 days after their pups were weaned, 48 h water-deprived males, males given 2% saline solution (dehydrated) for 10 days, males given 2% saline as described then given tap water to rehydrate for 2 or 5 weeks. Morphometric analysis of electron micrographs revealed that all stimuli leading to increased hormone release were accompanied by both increased occupation of the basal lamina by nerve terminals as well as decreased enclosure of neurosecretory processes by pituicyte cytoplasm. Neural occupation of the basal lamina remained significantly elevated 10 days post-weaning and at 2 weeks (but not 5 weeks) of rehydration following 10 days of dehydration. Pituicyte enclosure of neurosecretory axons had returned to control values in the postweaning and 5 week (but not 2 week) rehydrated animals. The mean length of individual nerve terminal contact with the basal lamina was found to increase under some, but not all, conditions associated with increased hormone release (i.e. parturition, acute and chronic dehydration, but not during lactation) and to decrease below control values in prepartum females and after 5 weeks of rehydration. During lactation, in contrast, the greater neural coverage of the basal lamina was accompanied by an increased number of terminals per unit length of basal lamina without an increase in the average contact length of the individual terminal.These data provide further evidence that neurohypophysial nerve terminal contact at the basal lamina and the relationship of pituicytes to neurosecretory processes can change under physiological conditions in a manner that is probably functionally related to controlling the contact area for hormone release.     

Post-natal development of the median eminence of the guinea pig

The ultrastructure of the median eminence of neonatal (newborn, 1-, 3-, 5-, and 10-day old) and adult guinea pigs was studied to determine the dynamic changes occurring in this structure during early life. At birth the portal vasculature consists of the Mantelplexus and a few, non-fenestrated capillary loops. The number of ansae and the degree of fenestration increase rapidly after birth. The abundance of cytoplasmic and ciliary projections into the ventricular recess and the large numbers of organelles indicate that the ependymal cells are more active in the neonatal period than in the adult male. Moreover, the ependymal endfeet cover most of the surface area of the primary portal plexus during this time. The neuronal layers of the median eminence are difficult to distinguish at birth due to the lack of myelinated fibers in the zona interna. Significant myelination appears on day 3 but is not complete until day 10. There is a progressive increase in the numbers of Herring bodies and large neurosecretory granules (1,500-1,700 Å) during this same time period. In the zona externa, few nerve terminals abut on the perivascular space until day 3. Increases in numbers of granules per axon profile were noted for each day after birth. Despite the relatively long gestation period of the guinea pig (68-72 days), the morphologic appearance of the median eminence at birth suggests that the neurovascular link controlling anterior pituitary function is not yet complete.     

Proliferation of capillary endothelial cells in the primary plexus of the hypophyseoportal system in rats during ontogeny

A light-microscopy study of primary portal plexus formation in rats between day 14 of prenatal life and postnatal day 9, carried out using thymidine autoradiography, showed that the formation and growth of capillaries for this plexus occurs through proliferation of endothelial cells whose mitotic activity is highest on days 14–16 of prenatal development. Endothelial cell differentiation and capillary development are characterized by flattening of the endothelium, widening of capillary lumens, and the formation of capillary loops that penetrate into the median eminence. The findings of this study indicate that capillaries of the primary portal plexus mainly develop in rats during the perinatal period. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 5, pp. 462–464, May, 1995 Presented by I. B. Zbarskii, Member of the Russian Academy of Medical Sciences     

Light- and electron-microscopic immunocytochemistry of glutamic acid decarboxylase (GAD) in the basal hypothalamus: Morphological evidence for neuroendocrine γ -aminobutyrate (GABA)

GABAergic cells and axon terminals were localized in the basal hypothalamus of different species (rat, mouse and cat), by means of an immunocytochemical approach using a specific and well-characterized antiserum to the GABA biosynthetic enzyme, glutamate decarboxylase. Lightmicroscopic visualization was performed with an indirect immunofluorescence method and electron-microscopic observations were made on material with pre-embedding staining and use of the peroxidase-antiperoxidase procedure.At the light-microscopic level, a dense immunofluorescent plexus was observed over both the medial and lateral parts of the external layer of the median eminence. The labelling extended from the rostral part of the median eminence up to the pituitary stalk. Over the subependymal and internal layers only a few immunoreactive dots were visible, except around the blood vessels where they appeared more concentrated. Immunoreactive varicosities could be found following the outlines of the capillary loops and lining tanycyte processes, especially in the median eminance midportion.At the electron-microscopic level, the immunolabelling was exclusively found over neuronal profiles in the median eminence. The latter represented a small fraction of the total number of varicosities visible on the same section. Labelled profiles typically contained numerous small clear synaptic vesicles and only a few or no dense-core vesicles. In the subependymal and internal layers, rare labelled endings were found close to ependymal cells or among transversally cut fibers, respectively. In the palisadic zone, elongated positive boutons were visible intermingled with bundles of unlabelled axons and glial or ependymal processes. In the neurohemal contact zone, immunoreactive endings were observed among unlabelled neurosecretory endings in close vicinity to fenestrated capillary perivascular space.Small moderately intense immunofluorescent varicosities were observed all over the hypothalamus. The density of the glutamate decarboxylase-positive network was higher than in most diencephalic regions. Intraventricular or topical injection of colchicine allowed the visualization of small lightly immunoreactive cells in the diffusion area of colchicine. In the arcuate nucleus labelled axonal endings containing small pleomorphic synaptic vesicles and sometimes a few dense-core vesicles were observed at the electron-microscopic level. Typical synaptic junctions were commonly found between positive endings and unlabelled perikarya, or more frequently, unlabelled dendrites.These findings show that glutamate decarboxylase-containing endings are localized in several strategic sites for potential GABAergic neuroendocrine regulations. The GABAergic endings found among neurosecretory endings in the neurohemal contact zone may provide the morphological support for the release of γ -aminobutyrate into the portal blood flow as an hypothalamic hypophysiotropic hormone. Alternatively, neurosecretory cells might be under GABAergic control expressed either at their terminal level within the median eminence or the cell body level within the parvicellular hypothalamic nuclei.