Development of the choroid plexus anlage and supraependymal structures in the fourth ventricular roof plate of human embryos: scanning electron microscopic observations

The developing anlage of the choroid plexus and supraependymal structures in the fourth ventricular roof plates of nine normal human embryos ranging from Carnegie stages 14 to 19 were investigated with scanning electron microscopy. In the human embryos at stage 18, the first semimacroscopic choroidal anlage developed in the form of bilateral evaginations that ran dorsomedially and caudally from the bilateral corners of the rhombencephalon. The anlage became evident with even smaller and parallel ridges in the embryo at stage 19. Embryos at earlier stages exhibited surface membrane modifications such as convexity, microvilli, cilia, and spherical protrusions at the middle one-third of the rhombencephalon, which corresponded to the future choroidal anlage region. Two morphologically different groups of supraependymal cells (SE cells) were elucidated throughout the stages examined. Type 1 SE cells has spindle or tear-drop-like bodies, frequently with one or more long cytoplasmic processes. Type 2 SE cells were globular, with numerous fine pseudopodial processes. Type 1 SE cells were distributed mainly at the future choroidal anlage regions or on the anlage itself and were less frequently located at the rostral end of the roof. We found no general pattern in the distribution of type 2 SE cells. Supraependymal fibers (SE fibers) were seen as fine processes that were distributed similarly to type 1 SE cells and extended transversely for a long distance.     

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     

The ependymal surface of the fourth ventricle of the rat: a combined scanning and transmission electron microscopic study.

The morphological features of the ependymal surface and supraependymal elements of the fourth ventricle of the rat were examined by scanning electron microscopy (SEM) and by the transmission electron microscopy (TEM). The results confirm the following aspects: 1) The presence of supraependymal elements and microvilli in the ependymal territories, including the sites where the cilia completely cover the ependymal surface; 2) The existence of cilia with oval or spherical thickenings together with supraependymal bulbs similar in size to those of the larger ciliary swellings; 3) Identification of the long supraependymal fibres with intermittent fusiform dilations observed under the SEM with the nerve fibres seen under the TEM; 4) The existence of intraventricular axodendritic synapses.     

The structure of the hypothalamic inferior lobes of the blacktip reef shark: Scanning and transmission electron microscopic observations

The inferior lobes of the shark hypothalamus were examined with light, transmission and scanning electron microscopy. The cells bordering the floor of the lateral recess appear to be typical liquor-contacting neurons. With scanning electron microscopy (SEM) the apical ends of these cells are seen to bulge into the ventricular lumen. In contrast, the roof is lined by a more typical ependymal cell characterized by numerous cilia and microvilli. In addition, SEM reveals several kinds of supraependymal cells with processes that appear to penetrate the ventricular lining. A periventricular nucleus underlies the ependymal cells. Neurons of the periventricular nucleus contain numerous lipofuchsin granules. The rest of the inferior lobe consists of many neuronal fibers. The morphology of the hypothalamic inferior lobe is discussed in relation to its possible role in feeding and aggressive behavior in both elasmobranchs and teleosts.     

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。     

Supraependymal cells occurrence in ventricular system of small ruminants. Scanning electron microscopy study

The third cerebral ventricle ependymal lining including eminentia medians was studied by means of SEM in 20 sheep, 13 goats and one goat hermaphrodite. Supraependymal cells in addition to the usual supraependymal structures were observed. In sheep, they occurred in the infundibular low part only in females during oestrus. In goats, they were present in almost every case with the exception of male animals during the "rest" period (April). The number, topography and-to some extent--the appearance of the goat supraependymal cells were in relation to the animal's sex, and the females to the ovarian cycle phase. The supraependymal cells were found on the eminentia mediana surface only in the goats. In small ruminants, the processes of most supraependymal cells formed the ruffled membranes and only the eminentia mediana supraependymal cells--and in hermaphrodite also in the infundibular rostral part--resembled more neurons.     

Scanning electron microscopic analysis of the linings of the fourth ventricle in the domestic fowl

Summary Surface features of the ependymal linings of the fourth ventricle in the fowl were analyzed employing the scanning electron microscope (SEM). On the floor of the median sulcus, each ependymal cell has a solitary cilium, whereas on both sides of the sulcus, cilia are so densely distributed that the details of the underlying cell surface are usually obscured. On the roof of the fourth ventricle, except for the surface of the ciliated groove where numerous cilia are present, the ependymal cells are polygonal in shape, and the center of each cell possesses an aggregate of ten to twenty cilia. Cell surfaces of the choroid tela are entirely covered with delicate microvilli and possess clumped cilia. The ependymal cell surfaces of the area postrema are dome-like in shape. Each ependymal cell has a solitary cilium and shows a smooth surface free of microvilli.This work was supported by a Scientific Research Grant, No. 144017, from the Ministry of Education of Japan to Professor M. Yasuda     

Ultrastructural studies on the ventricular surface of the frog cerebellum

Summary Ultrastructural studies of the ventricular surface of the frog cerebellum showed regional differences. In the midline region of the adult cerebellum was found a band of profusely ciliated squamous ependymal cells. In the rest of the cerebellum the ependymal cells were columnar and each had a single cilium. In the cerebellum of the premetamorphic tadpole, the squamous ependymal cells of the midline region also were monociliated. During metamorphosis they gradually became multiciliated. Additionally, supraependymal cells and synaptic elements were present on the ventricular surface of the cerebellum of adult frogs as well as in late metamorphic tadpoles. In contrast, supraependymal cells were rarely observed in premetamorphic tadpoles, and it was concluded that the supraependymal system develops during metamorphosis. It is postulated that the band of cilia may be associated with the circulation of cerebrospinal fluid, and supraependymal synaptic elements function in neuroendocrine regulation.     

Ependyma and supraependymal structures in some areas of the fourth ventricle in the rat

The ependyma was investigated in five areas of the rat ventricle system by means of both light and electron microscopy. The columnar, cuboidal and flattened types of the ependymal cells were mainly seen. All of them were seen in the fourth ventricle, while in the aqueductus cerebri and in the central canal the flattened type of the cell was lacking. An unusual variation as to the form of the ependymal cells was found on the roof of the fourth ventricle. Three groups of intraventricular structures were found in all investigated parts of the ventricle system: supraependymal globular structures containing irregularly arranged cristae, supraependymal protrusions appearing as homogeneous contents, and nerve profiles including nerve endings and nerve axons. The morphological characteristics of the ependyma and intraventricular profiles in the fourth ventricle allow to suppose a certain role of these structures in the exchange of various materials between the CSF, ependyma and neuropile.     

Correlative scanning-transmission electron microscopic examination of the perinatal rat brain

Summary The ultrastructural organization of the perinatal hypothalamus and the dynamics of neuronal and ependymal growth and plasticity were examined in this investigation. The brains of fetal rats 16, 17 and 18 days in utero and those of postnatal rats 1–16 days post partum were fixed with aldehyde fixatives and prepared for combined SEM/TEM analysis. By day 17 in utero the ventricular (ependymal) surfaces of the fetal thalamic wall, cerebral vesicle and rhomboid fossa were relatively well differentiated with cilia and microvilli. Type II histiocytes were the first supraependymal cell to appear upon the ventricular lumen and were evident by day 17 in utero. In contrast, the apical surfaces of tanycytes of the infundibular recess as well as those of most other circumventricular organs were poorly differentiated and unremarkable. Tanycytes of the infundibular recess exhibited a simple hexagonal mosaic pattern of apposed plasmalemmata and even by day 1 post partum few cilia or microvilli were evident.By day 5–6 post partum Type I supraependymal neurons and axonal processes began to make their appearance with some emerging from the underlying parenchyma of the median eminence. By day 16 post partum the ventricular surface of the infundibular recess was comparable with that of the adult.The Type I supraependymal neurons are remarkably similar in their ultrastructural organization with parvicellular neurosecretory neurons elsewhere in the endocrine hypothalamus. Their emergence at day 5–6 post partum suggests a possible correlation with the critical period of sexual differentiation and a potential receptor role for this cell line. On the contrary this phenomenon may simply be a developmental anomaly. Nonetheless, the mergence of such elements upon the lumen of the third cerebral ventricle underscores a remarkable degree of neuronal plasticity in the perinatal hypothalamus.Supported by USPHS Program Project Grant NS 11642-04 and USPHS-BRSG Grant RR-05403.The authors wish to thank N. Kutryeff for her excellent technical assistance     

Morphology and distribution of supraependymal cells in the third ventricle of the albino rat

The supraependymal cells (SEC) are a normal component of the wall of the cerebral ventricles. In the hypothalamic area of the third ventricle they are restricted, in healthy animals, to the ependymal projection of the hypophyseotropic area. Here the SEC show great polymorphism. In addition to bipolar, multipolar and stellate or spider-like cells, transitional forms between these types can be seen. Their perikarya and processes can either remain at some distance from the ependyma or be in close contact with it. The processes may protrude between the ependymal cells or show surface differentiations that resemble the thin cytoplasmic folds of the mesenchymal wandering cells. Considering this and the variations in the number of cells, for example during the ovarian cycle, the SEC can be interpreted as mesenchymal cells, probably related to microglial cells of the subependymal layer. It is suggested that the SEC have a phagocytotic function and may be involved in the normal renewal of the ependyma. A definitive explanation for the restriction of the SEC to the hypophyseotropic area as well as the elucidation of their function remain to be found.     

Morphology of ventricular wall surfaces following neurotoxin induced degeneration of supraependymal afferents

Following intraventricular injection of 5,7-dihydroxytryptamine in rats and survival periods up to 25 weeks, the supraependymal serotonergic axon plexus in the lateral and fourth ventricle, except for the plexus upon the hippocampal fimbria in some cases, showed no evidence of regenerative capacities. This contrasts with many reports on regeneration of intraparenchymal serotonergic fibre systems in the mammalian brain following mechanical or neurotoxin-induced experimental degeneration. The supraependymal plexus appears to be critically involved in maintainance of normal ependymal integrity in that following its experimentally induced death many ependymal cells exhibit various pathological alterations in all regions examined. Degeneration of this plexus is also associated with heavy phagocytotic reactions. Morphology, distribution and possible origins of supraependymal macrophages are briefly discussed.     

Supraependymal cells and fibers in the third ventricle of the domestic chicken. A scanning electron microscopic study

Supraependymal cells, fibers and what are presumed to be neuronal bulb-like projections were found in the third ventricle of the domestic chicken with a scanning electron microscope. At least two types of supraependymal cells were found: neuron-like cells and phagocyte-like cells. The former were predominantly seen in the area of the paraventricular organ and infundibular recess. The latter were abundant on the ventricular surface of the median eminence and subfornical organ. Bulb or club-like projections thought to be the dendritic terminals of CSF-contacting neurons were observed in the area of the paraventricular organ and infundibular recess. Similar structures were observed at the preoptic recess as well. The supraependymal neuronal components found in the domestic chicken differed from those of mammals in several respects: 1. the wall of the third ventricle was devoid of supraependymal fibrous plexus except for that of the paraventricular organ; 2. bulb-like projections were abundant in the area of the paraventricular organ; 3. supraependymal neuron-like cells were unipolar or bipolar in appearance. These data underline the dissimilarity of the CSF-contacting neuronal system of birds and mammals.     

Scanning electron microscope observations of the rat subcommissural organ.

Under the scanning electron microscope, the rat subcommissural organ (SCO) appears as an oval zone, rich in kinocilia and well delimited from the non-specific ependymal epithelium. This zone surrounds the cranial and posterior part of the mesencephalic canal's entrance. The ependymal cells of the SCO show coniform processes with microvilli and kinocilia. In contact with the apical pole of the peripheric SCO-ependymocytes lie scarce supraependymal axons. The Reissner's fiber is composed by the twining of the fibrillary structures emerging from the area of the SCO.     

Scanning and transmission electron microscopy of intraventricular dendrite terminals of hypothalamic cerebrospinal fluid contacting neurons in Triturus vulgaris

A scanning (SEM) and transmission electron microscopic (TEM) study of the ventricular wall of the hypothalamus of Triturus vulgaris was performed with special regard to the intraventricular dendrite terminals of the cerebrospinal fluid (CSF) contacting neurons of the preoptic area (magnocellular and parvocellular preoptic nuclei), the infundibular lobe (anterior periventricular nucleus, infundibular nucleus), and the paraventricular organ. In the preoptic area and infundibular lobe, the terminals were knob-like or club-shaped, of various sizes (diameter about 0,5 to 3,0 micrometer) and located immediately above the ependyma. Ultrastructurally, they may contain dense-core vesicles of varying sizes. The CSF contacting dendrite endings of the paraventricular organ built up a supraependymal labyrinthic layer which could be divided into a rostral crest-like part and a caudal flat and broad division. In both parts, three main types of terminals of various size and shape could be distinguished: a) ramifying, b) elongated, and c) bulb-like dendrite endings which also differed by their TEM structure. The bulk-like terminals, first of all the small ones, originated from the distal part of the nucleus of the organ (nucleus organi paraventricularis) while the other two types took their origin from its intra- and subependymal part. In all areas investigated, each intraventricular dendrite ending gave rise to a solitary cilium (type 9 X 2 + 0). It differed from the ependymal kinocilia by both SEM and TEM characteristics. In the paraventricular organ, the neuronal cilia were hidden inside, or below the supraependymal layer of terminals. There were intraventricular axons which formed synapses on CSF contacting dendrite endings of both parts of the paraventricular organ. Free intraventricular neurons, further ependymal areas heavily or scarcely ciliated, were described. The CSF contacting dendrite terminals were predominantly present near ventricular recesses and in regions where the ependyma was scarcely ciliated.     

Ependymal foldings and other related ependymal structures in the cerebral aqueduct and fourth ventricle of man

The ependymal lining of the cerebral aqueduct and fourth ventricle of 100 normal humans was studied with the light microscope. Ependymal foldings with normal morphology and a constant distribution pattern were detected in all. The most common sites were the median sulcus and sulcus limitans in the fourth ventricle, and the ventral and lateral walls in the cerebral aqueduct. Rows, islands and rosettes of ependymal cells embedded in normal subependyma were present in 25/82 adults (30%) and in 3/18 children (16%) in a similar distribution pattern as that of the ependymal foldings. We illustrate these normal structures which probably result from fusion between the walls of the ependymal foldings and distinguish them from granular ependymitis and postmortem artifact.     

Ultramicroscopic organization of the vascular plexus in the rabbit cerebral lateral ventricles and its changes under the effect of pyrogens

By means of electron microscopical methods organization of various components of the vascular plexus in the rabbit cerebral lateral ventricles--ependymal and supraependymal cells, as well as capillaries, making the base of the hemato-encephalic and hemato-liquor barriers of the plexus have been studied. Injection of bacterial pyrogen (pyrogenal) to the animals is accompanied with an increasing permeability in the barriers of the vascular plexus for certain blood cells (lymphocytes, monocytes), their activation and transformation into plasma cells and macrophages. Under pyrogenal effect in the ventricle cavities activation of the supraependymal cells and intensification of their interaction with the underlying ependyma take place. In the ependymal layer local dilatation of intercellular spaces and intensification of exocytic processes are observed. The changes revealed demonstrate an active reaction of some elements of the vascular plexus to fever, dependent on injection of the bacterial pyrogens into the organism.     

The collicular recess organ: Evidence for structural and secretory specialization of the ventricular lining in the collicular recess

Summary The collicular recess organ and adjacent portions of the collicular recess were studied by light microscopy, scanning electron microscopy and transmission electron microscopy. In the collicular recess, the ventricular wall contains folds and is well vascularized. The adluminal ependymal cells generally bear kinocilia and microvilli on their ventricular surface. Among the cilia, many secretory droplets, some axons, and few supraependymal cells are seen. Various stages of apocrine ependymosecretion are observed. In addition to tanycytes, coelocytes are found scattered throughout the ependymal lining of the collicular recess. Coelocytes, characterized by lumina containing cilia and a few microvilli, are accumulated in ependymal and hypependymal positions of the collicular recess organ at the roof of the collicular recess.Supported by PHS grants NS 09914 and T32 CA09156. We thank Dean Wyrick and John McNeill, Jr., for their technical assistanceNRSA Postdoctoral Trainee     

Cytochemical localization of alkaline phosphatase in the ependyma of the rat medulla oblongata

Summary The ependyma of the IVth ventricle and the central canal of the rat medulla oblongata was investigated using the cytochemical technique for alkaline phosphatase (AlPase) which revealed two types of ependymal cells in the medulla. The central canal type of the ependymal cell occupying the dorsal part of the central canal in the lower medulla exhibited intense AlPase activity with light microscopy. These cells had reaction products in all plasma membranes, including the microvilli and the cilia at the luminal cell surface. Some cells appeared to be tanycytes, since the process reached the basement membrane of the parenchymal blood vessel. The ventricular type of ependymal cells, which form the floor of the IVth ventricle and the central canal, contained no reaction products in any structure of the luminal cell surface.The possible relationship between the cerebrospinal fluid and the nervous tissues through the ependymal linings is discussed.     

Thrombin causes the Enrichment of Rat Brain Primary Cultures with Ependymal Cells Via Protease-Activated Receptor 1

Ependymal cell culture models from rat have been developed over the last 20 years to facilitate biochemical studies on this least-studied glial cell type. The cell culture protocol calls for the presence of thrombin, which is essential for obtaining a high proportion of multiciliated ependymal cells. The serine protease appears to act via protease-activated receptor 1 to prevent the apoptosis of ependymal precursors and enhance their proliferation without affecting contaminating cells. Unciliated precursors differentiate into polyciliated ependymocytes by passing through a stage of monociliation. The message for protease-activated receptor (PAR) 1 is initially abundant in the cultures, but its level declines as the cells differentiate. Besides PAR 1, signalling through PAR 2 also promotes ciliation in rat brain primary cultures, albeit to a lesser degree than the thrombin receptor. Thrombin and other proteases may be involved in the regulation of ventricular wall development. This action would be mediated mainly by PAR1.