The ventricular surface of the area postrema and the adjacent area subpostrema in the rabbit brain]

The ependymal surface of the area postrema (rabitt) was examined by scanning and transmission electron microscopy. The flattened ependymal cells show few microvilli. Towards the central canal, the ependymal cells change gradually to a columnar shape; the number of microvilli increases concomitantly. The area postrema ependymal cell surface mostly bears a single cilia. In contrast, a region immediately adjacent to the area postrema, which has been named area subpostrema (Gwyn and Wolstencroft 1968), shows cilia arranged in bunches. These cilia are regularly covered with colloid -- like droplets. A period-acid-bisulfit-aldehydthionine method (Specht 1970) permits to identify these droplets with glyproteids.it has been suggested that the droplets might derive from the area subpostrema ependymal cells. Above the ependymal surface of the area postrema, a great number of fine unmyelinated neuronal processes and thicker processes are observed. Some of them show bulb-like endings. These terminals contain small vesicles, dense cored vesicles (400...800 A), and mitochondria which are mostly characterized by a single central prismatic tubule. The plasmalemma of some bulbs is in a synaptic contact with the apical plasmalemma of the ependyma, while other bulbs see to end freely in the ventricle. Some neuronal processes penetrate between ependymal cells of the area postrema into the ventricular lumen.     

Fine structure of the ependyma and intercellular junctions in the area postrema of the rat

Summary Ependymal cells and their junctional complexes in the area postrema of the rat were studied in detail by tracer experiments using horseradish peroxidase (HRP) and colloidal lanthanum and by freeze-etch techniques, in addition to routine electron microscopy. The ependyma of the area postrema is characterized as flattened cells possessing very few cilia, a moderate amount of microvilli, a well-developed Golgi apparatus and rough endoplasmic reticulum. Numerous vesicles or tubular formations with internal dense content were found to accumulate in the basal processes of ependymal cells; the basal process makes contact with the perivascular basal lamina. It is suggested that the dense material in the tubulovesicular formations is synthesized within the ependymal cell and discharged into the perivascular space. The apical junctions between adjacent ependymal cells display very close apposition, with a gap of 2–3 nm, but no fusion of adjacent plasma membranes; they thus represent a transitional form between the zonulae adhaerentes present in the ordinary mural ependyma and the zonulae occludentes in the choroidal epithelium. A direct intercommunication between the ventricular cerebrospinal fluid (CSF) and the blood vascular system indicates that a region exists lacking a blood-ventricular CSF barrier.     

The ependyma of the ventriculus mesencephali in the rat

The ventriculus mesencephali in the rat proceeds in the colliculi posterior region from the mesencephalic aqueduct in the dorsal direction. The ependymal lining is formed by flat, cuboid and cylindrical cells. The cylindrical cells, which occur in all parts of the ventriculus mesencephali, are the most numerous. The cuboid cells are localized mainly in the anterior part, the flat ones in the posterior part of the ventriculus mesencephali. Some cuboid and cylindrical cells have short basal processes. Among the ependymal cells, there are cells with long basal processes. The ependyma is at sites interrupted by flocks of ependymal cells. On the cell surface, there area cilia, microvilli and protoplasmic extrusions. The cell nuclei are spherical to oval. Supraependymally, there are homogeneous globules and intraventricular fibres. The histological variability of ependymal cells may point to the active participation of these cells in functional processes, even within such a small part of the ventricle system as the ventriculus mesencephali.     

Morphological and neurochemical diversity of neuronal nitric oxide synthase-positive amacrine cells in the turtle retina

By gently scraping off the surface of the lateral ventricles of adult bovine brains, we obtained sheets containing the ependymal layer and some attached sub-ependymal cells. Explants were cultured in serum-free medium or in two media enriched with 20% fetal calf serum or 20% adult bovine cerebrospinal fluid, and processed for different time intervals from 4 h to 60 days. For characterization of the ependymal cells we used antisera against S-100 protein, vimentin and glial fibrillary acidic protein (GFAP). For comparison, the ependyma of adult bovines and of fetuses from days 60 to 120 post coitum was studied in situ. The adult ependyma consisted of a ciliated, cuboid cell monolayer with short basal processes; it displayed S-100 immunoreactivity but only scarce deposits of vimentin and no GFAP. The fetal ependyma had the appearance of a pseudostratified epithelium with elongated nuclei and basal processes containing S-100 and vimentin from day 80 post coitum and GFAP from day 100 post coitum. In explants, no differences were seen between the three culture media; the ependyma became pseudostratified, developed basal processes and showed increasing amounts of S-100 and vimentin first, and subsequently also GFAP. These changes were concomitant with the onset of mitotic activity in the subependymal layer leading to the production of numerous cells. The morphological and immunocytochemical features of ependymal cells in cultured explants resembled those of fetal ependyma. Our results indicate that the culture of ependymal explants from adult bovine lateral ventricles is an useful model system for morphological and functional studies of the ependyma and for the analysis of cell proliferation in the subependymal layer.     

Ependyma and ependymal protrusions of the lateral ventricles of the rabbit brain

Summary The ependymal lining of the lateral ventricles of the rabbit brain was studied by means of scanning (SEM) and transmission electron microscopy (TEM). There exist cells devoid of cilia in the anterior horn over the region of the caudate nucleus, in the inferior horn over the hippocampus and on the opposite side over cortical regions. On the surface of some of these ependymal cells, accumulations of cytoplasmic folds and globules can be found. They bulge at different height over the ependymal cells. Clots of these cell particles are tied off from the cell, coming to lie as globules either on or between the cilia of the ependyma. TEM reveals that these tissue protrusions are cell debris consisting of different sized vesicles, cell organelles, tubuli and filaments. They originate from the ependymal layer but may reach down to subependymal cells. Multivesicular protrusions into the ventricular lumen are also observed. Possible causes of these protrusions are discussed; they are likely to be related to the age of the animals.On the ependyma of the caudate nucleus cilia, microvilli, microblebs and supraependymal neuronal cell processes are distributed unevenly over the surface. Within regions where cilia predominate there are cells which are tightly covered with microvilli. A certain direction of the course of the supraependymal neuronal fibers could not be found.The author is pleased to acknowledge useful discussions with Prof. Dr. med. E. van der Zypen. This study was partly supported by the Stanley Thomas Johnson Foundation     

Neurosecretory processes projecting from the preoptic nucleus into the third ventricle of Zoarces viviparus L.

Summary The cellular processes loaded with neurosecretory elementary granules penetrate the ependyma and project into the third ventricle at the level of preoptic nucleus of Zoarces viviparus L. These cellular processes seem to be arising from the neurosecretory cells. At this zone, not all but some of the ependymal cells have a cilium. In this paper the possible function of these neurosecretory processes and the ciliated ependymal cells are discussed.This work was aided by a Grant from NATO and the Deutsche Forschungsgemeinschaft.     

Enzymatic organization of the subcommissural organ.

In the subcommissural organ (SCO) of the guinea pig, rat, golden hamster, and mouse the activity and distribution of enzymes related to the energy-supplying metabolism and of some marker enzymes of different cell organelles have been investigated by means of mostly modified histochemical methods. The results were compared with findings in the ciliated ependyma of the ventricular wall and with those in the ependyma of the choroid plexus of the third ventricle. In the ependymal part of the SCO only a moderate activity of hexokinase is observed in its specialized columnar cells whereas a high activity is present both in the ciliated ependyma and the choroid plexus. - The staining pattern of glucose-6-phosphatase is similar to that of hexokinase but this enzyme is found is the SCO only. - Likewise hexokinase, glycogen granules and enzymes related to glycogen metabolism (phosphoglucomutase, uridine-diphosphoglucose pyrophosphorylase, glycogen synthetase and phosphorylase) are regularly found most numerous and active in the nuclear and supra-nuclear area of the ependymal part. These enzymes are less active in both the other ependymal regions. - Uridine-diphosphoglucose dehydrogenase could not be demonstrated in the SCO. The NADP-linked enzymes of the pentose phosphate shunt, glucose-6-phosphate and 6-phosphogluconate dehydrogenase, show a moderate activity which decreases also from the nuclear towards the apical area of the ependymal cells of the SCO. Enzymes of the glycolytic pathway, such as glucosephosphate isomerase, fructose-6-phosphate kinase, fructose-I,6-diphosphate aldolase, glyceraldehyde-3-phosphate and lactate dehydrogenase, are highly active in the SCO and are located mainly in the supranuclear area, too. Fructose-1,6-diphosphatase could not be demonstrated thus indicating that in the SCO the pathway is most probably only glycolytic but not gluconeogenetic. Compared to the ependyma of the ventricular wall and of the choroid plexus, in the SCO the M type subunits of lactate dehydrogenase predominate. Glycolytic enzymes are also very active in the choroid plexus but less in the ciliated ependyma. Compared to the ciliated ependyma and especially to the ependyma of the choroid plexus, the activities of enzymes which are only present in mitochondria (NAD-linked isocitrate dehydrogenase, succinate dehydrogenase, NAD-linked malate dehydrogenase after preextraction, cytochrome oxidase, 3-hydroxybutyrate and glycerolphosphate and glutamate dehydrogenase) are relatively low. Mitochondria are accumulated near the superior pole of the nuclei as well as in the most apical part of the ependymal cells. - The staining pattern of NADP-linked isocitrate and malate dehydrogenase as well as of NADH dehydrogenase suggests that these enzymes are localized both in and out of mitochondria. The extramitochondrial activity of the first two enzymes might be localized in the cytosol. The extramitochondrial activity of NADH dehydrogenase might be localized in the endoplasmic reticulum...     

The growth and differentiation of the regenerating spinal cord of the lizard, Anolis carolinensis

The present work describes the ultrastructure of the spinal cord in the regenerating tail of the lizard, Anolis. The distal growing region of the tail contains the advancing ependymal tube which is relatively devoid of axons but already contains channels between ependymal cell processes which anticipate their ingrowth. More proximally, fascicles of naked axons having their origin in the stump are present in the ependymal channels. Therefore, the pattern of fiber regeneration in the spinal cord is prescribed by the ependyma and not by the growing axons. Details of the ultrastructure of proximal, intermediate, and distal regions of the regenerate are reported. Particular attention is paid to the structure and differentiation of the ependymal cells and the relation of the ependyma to other glial cells, to nerve fibers, and to meningeal tissues.     

大白鼠第三脑室室管膜的超微结构

本文用扫描和透射电镜证实在成年大白鼠的第三脑室存在室管膜上神经元样细胞、神经胶质细胞和类组织细胞。神经纤维发自神经元样细胞或自脑室外穿入室腔而来,其末梢内含有清亮囊泡或兼有大颗粒囊泡。室腔內尚有膨大的树突末梢和室管膜细胞的球状小体。上述各种结构与感受、分泌和调节功能有关,并为下丘脑控制垂体机能的另一新途径(经脑脊液和室管膜)提供了形态学依据。     

Distribution of vitamin D binding protein expressing neurons in the rat hypothalamus

We observed immunostaining for vitamin D binding protein (DBP) in rat hypothalamus. Part of the supraoptic and of the paraventricular neurons showed DBP immunoreactivity, in part colocalized with Arg-vasopressin. DBP was also observed in widespread axonal projections throughout the lateral hypothalamus, the median eminence and the posterior pituitary lobe. A portion of ependymal cells, the choroids plexus epithelium and some of the endocrine cells in the anterior pituitary lobe contained DBP immunoreactivity. In situ hybridization of semithin sections with a synthetic oligonucleotide probe to DBP mRNA resulted in staining of magnocellular hypothalamic neurons, but not of ependymal cells or anterior lobe cells. Our observations indicate an intrinsic expression of DBP in the rat hypothalamus. DBP may be synthesized and transported along with the classical neurohypophyseal hormones. The multiple locations of DBP-expressing neurons indicate multiple functional properties: DBP may be released from in the posterior lobe, it may act as a hypophyseotropic factor and as a central neuroactive substance.     

Fine structure of the rhombencephalic tela of the bullfrog,Rana catesbeiana

Summary The posterior rhombencephalic tela choroidea of the bullfrog was examined by electron microscopy. This membrane, the pia-ependymal roof of the caudal hindbrain, contains a large central region characterized by cuboidal ependymal cells which surround sizable microscopic apertures — the interependymal pores.Ultrastructurally ependymal cells of this area are characterized by infrequent apical microvilli and cilia. They contain irregularly shaped nuclei and few cytoplasmic organelles that are largely apical in position. The most striking feature is an abundance of cytoplasmic filaments forming an extensive cytoskeleton. Laterally these cells are joined by numerous elaborate desmosomes. The majority of the ependymal cells have a basal lamina consisting of single, double, or triple laminae lying parallel to the basal plasma membrane.Several unusual specializations are seen at the margins of the interependymal pores. The ependymal cells have lateral cytoplasmic processes that form the actual border of each pore. These processes originate from the apical surface of the cell and partially enclose an elaborate network of basal lamina associated with the interependymal pores.These findings demonstrate microscopic apertures in the roof of the fourth ventricle in the bullfrog that are associated with an unusual form of supportive ependyma.     

Autoradiographic study of the proliferative activity of the preoptic recess ependyma in yearling and juvenile common frogs

Proliferative activity of the ependyma, lining the recessus praeopticus in juvenile frogs was studied with 3H-thymidine radioautography. Usually much more pronounced proliferation of ependymal cells occurred in the preoptic region in one year old frogs as compared with two year old ones. It can be concluded that in the former animals the migration of postmitotic labeled cells into the subependymal zone of the recessus preoptic area is significantly more intense. By the 30th day after multiple isotope injections some newly formed neurosecretory cells with labeled nuclei were found in the 1-5 cellular position of the recessus praeopticus subependymal zone. It is postulated that in juvenile frogs the ependyma of lateral wall of recessus praeopticus is probably a source ("cambium"), from which some young neurosecretory cells may originate.     

Sexual differences in the ependyma lining the third ventricle in the area of the anterior hypothalamus of adult rhesus monkeys

Summary Previous studies have shown that a circumscribed region of the anterior hypothalamus of the rhesus monkey is lined by tanycyte ependyma and it has been suggested that this ependyma which links the third ventricle with the pars tuberalis may have a functional role in the hypothalamic regulation of anterior pituitary function (Anand Kumar and Knowles, 1967a). In view of the known sexual differences in the hypothalamic regulation of pituitary gonadotropin secretion the present investigation was made to determine whether any structural differences were evident in the tanycyte ependyma in male and female rhesus monkeys.The results of this investigation are based on light and electron microscopic studies of the hypothalamus in 24 rhesus monkeys comprising 12 adult females, 11 sexually mature males and a two month old sexually immature male.The tanycyte ependyma in the rhesus monkey is double layered. There are bulbous projections on the ventricular surface of the cells in the ependymal layer nearest to the ventricle (the first layer of ependyma). These bulbous projections vary in size in relation to the menstrual cycle. They are well developed during mid-cycle and regressed during menstruation. In the males, where the secretion of pituitary gonadotropins does not occur cyclically as in the females, there was no marked variation in the bulbous projections between different individuals as in the female monkeys.In the sexually mature males, but not in the females, the two layers of ependyma are separated by a distinct space. The absence of such a space in the sexually immature male suggests that this difference may be related to sexual maturity.In the adult males the cells in the ependymal layer below the first layer of ependyma have microvilli which extend into the space between the ependymal layers. In the females where such a space is not present, microvilli were not evident.The precise functional significance of the tanycyte ependyma is not known. It is hoped that the results of the present investigation would draw attention to the need for a more detailed examination of the physiological role of the tanycyte ependyma in relation to reproduction.The expenses for this investigation were met from a grant made by the Ford Foundation to Professor Sir Solly Zuckerman and the electron microscope was provided by the Medical Research Council. I am indebted to Sir Solly for his interest in this work.     

Some morphological and ultrastructural changes in the ependyma of the amputation stump during early regeneration of the tail in the lizard,Anolis carolinensis

Light and electron microscope studies indicate that the old ependyma just proximal to the plane of amputation in early lizard tail regenerates shows a sequence of morphological changes which suggests that it as well as the new ependyma growing into the regenerate may play an active role in the initiation and maintenance of early tail regeneration. The old ependyma close to the plane of amputation undergoes hypertrophy and/or hyperplasia causing a partial closure of the central canal and pseudostratification. Its nuclei shift from an original apical position to a basal one. The ependymal processes become more prominent and extend to the pia, a condition not found more rostrally. There is also a significant increase in the amount of Golgi substance and a moderate increase in the rough endoplasmic reticulum. These observations lead to the thought that these cellular changes may be an expression of enhanced secretion and other activities in the old and new ependyma just proximal or distal to the plane of amputation.     

Formation of multiple ependymas in the regenerating optic lobe of larvae of the Egyptian toad, Bufo regularis Reuss

In the regenerating optic lobe of Bufo regularis larvae, secondary ependymas were formed in both the dorsal part (optic tectum) and ventral region (tegmentum) of the lobe concerned. These secondary ependymas were frequently observed in the rostral and caudal tectal regions after complete excision of the tectum. Most of the multiple ependymal structures were formed by self-organization of groups of undifferentiated cells migrating from the primary ependyma lining the optic tectum. Others split off from the primary ependyma, but remained in contact with it. The observations emphasize the wide range of possibilities of the cells produced by the larval tectal ependyma in response to partial or total excision of the tectum. The results suggest that cells of ependymal origin, in regenerating tectum, are capable of self-organization to complete ependymal tubes in the absence of direct with the primary ependyma.     

Up-regulation of neural stem cell markers suggests the occurrence of dedifferentiation in regenerating spinal cord

Following tail amputation in urodele amphibians, an ependymal tube, that resembles a developing neural tube, forms from ependymal cells that migrate from the cord stump and elongates by cell proliferation. Expression of the keratin pair 8 and 18 has been observed in the developing urodele nervous system and is maintained in the ependymal cells of the mature cord. We show here that expression of these keratins is not unique to urodeles, but is also observed in the radial glia of the human spinal cord, suggesting that these proteins might play a role both in neural development and regeneration. Analysis of their expression in the regenerating spinal cord following tail amputation shows that their expression, as well as that of glial fibrillary acidic protein (GFAP), is maintained in the ependymal tube during regeneration, though differences in their levels of expression are observed along the anteroposterior axis and appear to be related to the progression of morphogenesis. In addition, we show that following tail amputation the ependymal tube expresses the neural stem cell markers nestin and vimentin, which are undetectable in normal urodele spinal cord. This up-regulation of neural stem cell markers shows that the ependymal cells undergo a phenotypic change. Whereas maintenance of keratin and GFAP expression in the adult ependyma may reflect a higher plasticity of these cells in adult urodeles than in other vertebrates, re-expression of markers of early neural development suggests the occurrence of a dedifferentiation process in the spinal cord in response to injury.Edited by J. Campos-Ortega     

Scanning electron microscopic observations of the ependymal cell and the subependymal layer of the third brain ventricle in the rat

This investigation was undertaken to clarify the three dimensional ultrastructure of the subependymal layer in relation with the ependymal cell layer in rat brain using the scanning electron microscope (SEM). The subependymal layer existing below the ependyma of the third ventricle in the brain of mature albino rats was examined with S E M. The hypothalamus freshly excised after median sagittal section was treated by collagenase with or without trypsin for a short while to remove the ependymal cells at the ventricular wall. After the enzymatic pretreatment of the specimen, many ependymal cells were removed and the subependymal layer was partially exposed. Most of the ciliated ependymal cells remaining at the ventricular wall extended long, single basal processes which then penetrated into the subependymal layer. The subependymal layer was composed of a delicate framework of thin processes of glial cells, ependymal cells and, in addition nerve cells. Scattered among the neuropil just beneath the ependymal cell layer, there were relatively small, globular subependymal cells. Occasionally, there were large bundles of unmyelinated nerve fibres in the subependymal layer. The individual nerve fibres distinctly showed many axonal varicosities within the fibres. Intermingled with the nerve fibres, glial processes of various forms were present. The structure of the ependymal cells and the subependymal layer was compared with the findings already reported in the studies using light and transmission electron microscope.     

Ependyma: phylogenetic evolution of glial fibrillary acidic protein (GFAP) and vimentin expression in vertebrate spinal cord

The phylogenetic evolution was studied of both glial fibrillary acidic protein (GFAP) and vimentin expression in the ependyma of the adult vertebrate spinal cord. Eleven species from different vertebrate groups were examined using different fixatives and fixation procedures to demonstrate any differences in immunoreactivity. GFAP expression in the ependymal cells showed a clear inverse relation with phylogenetic evolution because it was more elevated in lower than in higher vertebrates. GFAP positive cells can be ependymocytes and tanycytes, although depending on their structural characteristics and distribution, the scarce GFAP positive ependymal cells in higher vertebrates may be tanycytes. Ependymal vimentin expression showed a species-dependent pattern instead of a phylogenetic pattern of expression. Vimentin positive ependymal cells were only found in fish and rats; in fish, they were tanycytes and were quite scarce, with only one or two cells per section being immunostained. However, in the rat spinal cord, all the ependymocytes showed positive immunostaining for vimentin. The importance of the immunohistochemical procedure, the cellular nature of GFAP positive ependymal cells and the relationship between tanycytes and ependymocytes are discussed, as well as GFAP and vimentin expression.     

Preliminary electronmicroscopical observations on the ampulla caudalis and the discharge of the material of Reissner's fibre into the capillary system of the terminal part of the tail of Ammocoetes (Agnathi)

The discharge of the material of the Reissner's fibre (RF) and the massa caudalis (MC) into the "meningeal spaces" has until now not been studied in detail and with the aid of the electron-microscope. It was generally assumed that the material of the MC disintegrates in the "meningeal spaces", but the nature and function of those spaces have not been established. The CNS of Ammocoetes ist encompassed by the meninx primitiva, which is the meningeal tissue of the lower vertebrates; the meninx does not differentiate during ontogeny into the pia mater and the arachnoides. In contrast, the meninx primitiva of mammals is the anlage in the ontogenetical sense of the leptomeninges in adult individuals. The results presented in this paper are valid for Ammocoetes only; Petromyzon and Myxine must be studied anew from a perspective which may be the result of our study. Of particular interest would be an anatomical analysis of these caudalmost structures of the CNS (Ampulla caudalis [AC], MC, neuropori, lacunae etc.) in Branchiostoma. The diameter of the RF of Ammocoetes is approximately 1.7 micron; this is not different from the diameter of the RF in adult individuals. In the AC the RF divides into several smaller fibres which eventually disintegrate and form the amorphous MC. In sagittal sections, the fibrillary structure of the RF is distinct. In cross sections the small fibres may appear as globules. The terminal part of the RF is very often coiled. The ependyma of the caudal part of the canalis centralis (CC) ist not remarkable; liquor contacting neurons are frequently found with their dendrites touching the RF. Liquor contacting neurons are never found in the ependyma of the AC. The ependymal cells of the dorsal part of the AC possess neither kinocilia or microvilli. The disappearance of these organelles begins in the dorsal part of the caudalmost segment of the CC. Most surprising is the distribution of the attachment devices which are probably desmosomes. In the ependyma of the caudal part of the spinal cord the attachment devices are typically situated; in the ependyma of the AC these devices are absent. We use the general term "attachment devices" until the nature of these devices has been clearly determined. In the caudalmost part of the spinal cord as well as in the AC, the intercellular spaces between the ependymal cells communicate, forming a 3-dimensional labyrinth. If there are no attachment devices between the proximal parts of the ependymal cells, the intercellular spaces may provide a passageway between the CC and the loose tissue of the meninx primitiva.(ABSTRACT TRUNCATED AT 400 WORDS)     

Differential immunocytochemical staining for glial fibrillary acidic (GFA) protein,S-100 protein and glutamine synthetase in the rat subcommissural organ,nonspecialized ventricular ependyma and adjacent neuropil

Summary Antibodies raised against glial fibrillary acidic protein (GFA), S-100 protein (S100) and glutamine synthetase (GS) are currently used as glial markers. The distribution of GFA, S100 and GS in the ependyma of the rat subcommissural organ (SCO), as well as in the adjacent nonspecialized ventricular ependyma and neuropil of the periaqueductal grey matter, was studied by use of the immunocytochemical peroxidase-antiperoxidase technique. In the neuropil, GFA, S100 and GS were found in glial elements, i.e., in fibrous (GFA, S100) and protoplasmic astrocytes (S100, GS). The presence of S100 in the majority of the ventricular ependymal cells and tanycytes, and the presence of GFA in a limited number of ventricular ependymal cells and tanycytes confirm the glial nature of these cells. The absence of S100, GFA and GS from the ependymocytes of the SCO, which are considered to be modified ependymal cells, suggests either a non-astrocytic lineage of these cells or an extreme specialization of the SCO-cells as glycoprotein-synthesizing and secreting elements, a process that may have led to the disappearance of the glial markers.