Ultrastructural study of neurovascular contacts in the median eminence of the rat

Summary In the median eminence of the rat axons of the supraoptic-paraventricular-hypophyseal tract with elementary neurosecretory granules (150–200 m) traverse the internal zone. Terminals containing dense core vesicles 60–120 m in diameter end on the portal capillaries of the median eminence. A unique organisation of the primary portal capillaries is shown. Endothelial cells have many fenestrae. The pericapillary space has numerous extensions all of which represent a special zone around the endothelial tube. The fine structural organisation and function of the neurovascular contacts in the median eminence are discussed.     

Ultrastructural evidence for exocytosis in the median eminence of the rat

Summary Exocytosis has been demonstrated by electron microscopy in the external zone of the median eminence of the rat. Exocytotic profiles have been observed in nerve fibres characterized by the presence of granular vesicles with median diameters of 90–103 nm and agranular vesicles of about 50 nm. In addition to the small agranular vesicles, coated vesicles of the same size have been found in many nerve fibres, suggesting that at least part of the agranular vesicles in the median eminence originate by micro-pinocytosis. The nature of the fibres showing exocytosis is discussed. Attention is drawn to the possibility of identifying types of fibres in the median eminence by the occurrence of exocytosis.The technical assistance of Mrs. R. M. Y. Hartsteen is acknowledged with gratitude. We would like to thank Prof. J. Moll for his helpful criticism. We also thank Miss P. Delfos and Mr. W. van den Oudenalder for photographic assistance.     

Ultrastructural changes in the median eminence of the rat following deafferentation of the basal hypothalamus

Summary This report concerns a light and electron microscopic investigation of the median eminence and dorsal infundibular stem of the rat following surgical isolation (deafferentation). Using a modification of the Halász technique, the basal hypothalamus, including the arcuate nucleus and median eminence were surgically isolated from surrounding structures. Special attention was directed to the contact (external) zone of the median eminence and rostral infundibulum where tuberohypophyseal axons as well as ependymal cell processes abut upon the abluminal basement membrane of the portal perivascular space. The results of this study to date suggest that 9, 20, and 40 days following surgical isolation, there is a distinct increase in the population of tuberohypophyseal dense core vesicles. It is suggested that deafferentation abolishes inhibitory and excitatory input that serves to modify the cellular dynamics of tuberohypophyseal neurosecretory elements. Comments are also made on the presence of cistern-like structures in the lateral median eminence; the presence of vesicle-like inclusions in terminal ependymal processes is discussed in relationship to the role that ependyma may play in linking the third ventricle with the adenohypophysis.This research was supported by USPHS Grants NB 08171 and AM 10002. The authors are indebted to the excellent technical assistance provided by G. Krobisch Dudley. Further, the authors wish to express their thanks to Dr. Adolph Weindl for his valuable advice and criticism, and to Matilde Holzwarth for her helpful assistance.     

The tanycyte of the rat median eminence

Summary The present ultrastructural study proves the existence of nerve terminals closely apposed to the plasmalemmata of tanycytes in the rat median eminence. Several of these axo-tanycytic endings display remarkable accumulations of agranular endoplasmic reticulum in the form of pleomorphic vesicles which are closely apposed on either side of the plasma membrane of each cell compartment. Some of these vesicular profiles give the impression of structural continuity across both membrane systems. This phenomenon is discussed in the context of being a potential substratum for communication between both cell compartments.Supported by NIH Grant NS13717 and NSF Grant BNS 78-11820     

Ultrastructural characterization of exocytotic release sites in different layers of the median eminence of the rat

Summary The release of neuronal secretory products by exocytosis in different layers of the median eminence of the rat was investigated ultrastructurally after perfusion with Ringer solution containing tannic acid. Exocytotic images were observed in all layers studied. Neurohaemal release sites were found in the pars externa of the palisade layer, where they occurred not only against the basal lamina of the pericapillary space, but also opposite, adjacent to neuronal and glial elements. In the lateral portion of the pars externa of the palisade layer most release sites were separated from the pericapillary space or the pial surface by ependymal or glial processes. In the pars interna of the palisade layer, and in the reticular, fibre and subependymal layers, release was observed in different types of axonal processes without morphological synaptic specializations. We suggest that products released in the pars externa of the palisade layer are destined to reach the capillaries of the primary portal plexus. Although the non-vascular release sites may serve a similar hormonal function, they may alternatively represent the morphological correlate of axoaxonal contacts or of paracrine, non-synaptic release sites.     

Uptake of peroxidase from the third ventricle by ependymal cells of the median eminence

Summary Peroxidase injected into the subarachnoid space in mice is absorbed by ependymal cells of the median eminence. The ependymal cells of the median eminence of the rat and Japanese quail absorb peroxidase injected into the third ventricle. The processes of these ependymal cells terminate at the capillaries of the primary plexus or those surrounding the ventromedial nucleus of the hypothalamus. In all three species, peroxidase is absorbed by the ependymal cells of the paraventricular organ and by those in close proximity to it. Some ependymal cells send processes to the capillaries in the lateral nucleus of the hypothalamus. These phenomena are discussed in relation to adenohypophysial function.A part of this investigation was effected while the senior author held a Visiting Professorship at the University of Giessen [Department of Anatomy (Professor A. Oksche, Director)].     

Ultrastructure of the median eminence of the rat

Summary The ependymal cells bordering the median eminence to the third ventricle are characterised by many microvillus-like projections and bulbous cell processes of the luminal plasma membrane. The latter contain many vesicles 500–1,000 Å in diameter. Cilia with 9+2 fibrillar pattern are seen occasionally. Adhesive devices in the from of zonula adhaerens and zonula occludens are found in the apical part of the intercellular junction. Unmyelinated nerve fibres with a mean diameter of 1 and containing many electron dense granules of 830–1,330 Å are often seen between the ependymal cells.Two types of glial cells are found in the median eminence. One is characterised by a nucleus with dense blods of chromatin and dense cytoplasm, and it is associated chiefly with the nerve fibres in the region of the hypothalamo-hypophysial tract. The other type of glial cell is characterised by fine, uniformly distributed chromatin in the nucleus and a relatively pale cytoplasm and branched processes which terminate perivascularly in the base of the median eminence.Myelinated nerve fibres are seen only in the region of the hypothalamo-hypophysial tract. Only a part of them contain electron dense granules 1,330–2,330 Å in diameter.Three types of unmyelinated nerve fibres can be distinguished in the median eminence according to the size of the electron dense granules they contain: 1. Nerve fibres containing granules 1,330–2,330 Å in diameter. They are seen primarily in the hypothalamo-hypophysial tract, but also in the zona externa; 2. those containing granules with a mean diameter of 1,330 Å; and 3. those containing granules with a mean diameter of 1,000 Å. The last two types are both encountered in the hypothalamo-hypophysial tract, the zona externa and the perivascular region of the base of the median eminence. Under high magnification, the membrane of the granules show evidence of a trilaminar structure and the content of the granules with a low electron density appeares to consist of small microvesicles or globular components. Besides granules, these nerve fibres contain vesicles mostly 420 Å in diameter whose relative number increases towards the perivascular nerve endings. 53 per cent of the inclusions in the hypothalamo-hypophysial tract are granules and 47 per cent vesicles, while the corresponding percentages for the zona externa are 40 and 60 and for the perivascular nerve endings 20 and 80.The mean width of the pericapillary space is 1 , but it varies greatly. It containes many collagen fibrils and fibroblasts. The capillary endothelium is frequently fenestrated and contains many vesicles of various sizes.Two types of granules-containing cells are found in the pars tuberalis depending on the size of the electron dense granules: 1. cells containing granules with a mean diameter of 1,330 Å: and 2. cells containing granules with a mean diameter of 2,000 Å. In addition, there are occasional follicular cavities filled with amorphous material, microvilli and cilia of 9+2 fibrillar pattern.Aided by a grant from the Sigrid Jusélius Stifteise.     

Ultrastructural features of the lateral preoptic area,median eminence and arcuate nucleus of the rat

Summary The arcuate nucleus, median eminence, and the lateral preoptic area from the brains of aldehyde-perfused male and female rats were examined by electron microscopy. In the lateral preoptic area, three neuronal types are described: a small light neuron, a larger light one, and a dark neuron resembling the larger light one in size and nuclear shape. Many myelinated axons are interposed among single neurons or neuronal pairs. The relationship of structures to each other is discussed. Several observations not previously reported are illustrated from tissue of the arcuate nucleus and median eminence.     

Ultrastructure of the rat median eminence after superfusion

Summary Isolated medio-basal hypothalami of adult rats were continuously superfused in a chamber with controllable inputs and outputs, for periods from 30 to 240 min. The median eminence was prepared for transmission electron microscopy under carefully controlled conditions by immersion fixation with osmium tetroxide. The ultrastructure of superfused median eminence was compared with that of directly fixed, non-superfused median eminence. Even after 4h of superfusion, the median eminence displays remarkably well preserved histological and cytological patterns; cytomembranes, cell organelles, intercellular relationships, and extracellular spaces were remarkably similar in superfused and non-superfused tissues. As a consequence of osmium tetroxide fixation, microtubules were not observable. The ultrastructural information obtained from unstimulated rat median eminence superfused in vitro provides a basis for future morphofunctional correlations in the study of neurosecretion.     

Localization of serotonin within tanycytes of the rat median eminence

Summary Formaldehyde and glyoxylic acid histochemical methods were employed to examine monoamine fluorescence of the rat median eminence. Tanycytes of the median eminence contained a yellow histofluorescence which was verified with microspectrofluorometry as due to the presence of serotonin. Catecholamine-containing varicosities, arranged in linear profiles throughout the depth of the median eminence, were observed. These linear profiles appeared to follow the contours of serotonin-containing tanycytes. Organculture experiments supported the hypothesis that the serotonin associated with tanycytes is localized within the tanycytes and does not arise from an extrahypothalamic source of nerve terminals. These data provide evidence that a tanycytic catecholamine-indoleamine morphological juxtaposition occurs in a manner reminiscent of that of another circumventricular organ, the pineal.Supported by USPHS Grants NS11642 and AM-19761USPHS Career Development Awardee NS-00259     

Immunohistochemical localization of thyrotropin-releasing factor in the rat median eminence

Summary Antibodies raised in rabbits against a synthetic preparation of thyrotropin-releasing factor (TRF) were used in association with the immunoperoxidase histochemical technique to localize TRF in the rat median eminence. Using coronal sections cut through the mid-arcuate region, specific immunoreactive material was observed in the medial and lateral regions of the superficial layer of the external median eminence.This work was financed by a grant from the Medical Research Council of New Zealand     

Electron microscopic and pharmacological studies on the rat median eminence

Summary The rat median eminence contains at least three kinds of granules or vesicles: 1. large electron-dense granules (perhaps carriers of neurohypophysial hormones), 2. small electron-dense granules with or without haloes (perhaps carriers of catecholamines) and 3. synaptic vesicle-like structures (perhaps carriers of acetylcholine). The former two electrondense granules exist in separate axons but they coexist with the latter vesicles in the same axons.The pars nervosa shows basically a similar structure to the median eminence. However, the axons containing the small electron-dense granules are very few. In the pars tuberalis, there are at least two types of cells: the cells of one type contain much cytoplasm with large round nuclei and those of the other type contain a small amount of cytoplasm with polymorphic nuclei. The cells of the former include multivesicular bodies and secretory granules, but those of the latter do not. Some of capillaries of the primary plexus are surrounded by the cells of the pars tuberalis on one side and by neurosecretory axon endings on the other side.The median eminence contains high concentration of acetylcholine or an acetylcholine-like substance and shows neurohypophysial hormone activity.Aided by Grant A-3678 from the United States National Institute of Arthritis and Metabolic Diseases. The authors are indebted to Dr. Welsh, Harvard University, for the kind gift of Mytolon.     

Nitric oxide inhibits tyrosine hydroxylase of rat median eminence

Tyrosine hydroxylase (TH) is the rate-limiting enzyme in the biosynthesis of catecholamines. In a previous report we found that intracerebroventricular administration of nitric oxide (NO) generator sodium nitroprusside (SNP) to conscious male rats inhibited dose-dependently the TH activity of the median eminence (ME). In the present study we have tested the in vitro effects of SNP on TH activity, its possible mediator and action mechanism. Exposure of the ME TH to SNP (50, 100 and 500 microM) caused concentration-dependent inhibition of its enzyme activity. Addition of; reduced hemoglobin Hb (10 microM), a NO scavenger, superoxide dismutase SOD (1000 units/ml), a superoxide scavenger enzyme, or uric acid UA (300 microM), a peroxynitrite scavenger, did not affect the enzyme activity by themselves, but prevented the inhibitory effect of SNP 500 microM. However, the presence of methylene blue MB (100 microM), a guanylyl cyclase inhibitor, did not alter either basal enzyme activity or the inhibitory action of SNP 500 microM. These results suggest that this action of SNP on TH of the ME would be mediated by peroxynitrite generated by the reaction of NO with superoxide.