Some observations on the fine structure of the corpus striatum of the rat brain

Summary The rat corpus striatum was perfused vitally with glutaraldehyde, immersed in OsO₄ and then observed under an electron microscope.Numerous small cells in the neostriatum show a simple cytoplasmic structure, while the large cells possess a complicated fine structure. These are differentiated under the elctron microscope into two kinds, which seem to have functional differences. The large pallidal cells containing much pale cytoplasm are covered with many varied axonal boutons from the cell body to the dendritic terminal making numerous axo-somatic or axo-dendritic trunk synapses. Numerous axo-dendritic, or spine synapses are recognized in the neostriatal neuropil.These numerous axon terminals, which belong to striatal nerve cells or other nuclei of the brain, are classified morphologically into several types. At least five types of synaptic vesicles are distinguished by their size or by the presence of fine dense granules on their membranes, and seem to be specific to the neostriatum.Many myelin interruptions and several kinds of glial cells in the corpus striatum are observed. Moreover, the ventricular wall of the caudate nucleus, namely, the ependyma, and two kinds of subependymal cells are described and discussed with reference to the subependymal layer.     

Immunocytochemical characterisation of the wall of the bovine lateral ventricle

The cytoarchitecture of the walls of the bovine lateral ventricles was investigated by the use of immunocytochemistry. We defined three types of walls. Type 1 lined regions of white matter and had ciliated cuboidal ependyma, a few subependymal cells and a narrow subjacent glial layer. Type 2 lined the striatum and possessed ependymal cells with conspicuous basal processes that extended through a wide subependyma containing many subependymal cells and a wide subjacent glial network. Type 3 lined the rostral horn and displayed ependymal cells with the longest basal processes and wider subependymal and glial layers. Ependymal cells of type 2 and 3 walls were labelled with antibodies against S-100 protein, vimentin, GFAP, BLBP and nestin. Anti-III-tubulin stained small cells in the subependyma and inside the GFAP- and vimentin-positive subjacent glial network. Anti-PCNA-positive nuclei were abundant in the subependymal and glial layers of type 2 and 3 walls. DiI in vitro tracing studies revealed small bipolar cells in the glial layer at a distance from the site of the label deposit. These results suggest that neurogenesis takes place in adult bovine subependyma mostly in the walls of the striatum and the rostral horn, and that young neuroblasts may migrate in a rostro-ventral direction through the glial network.This work was supported by DGICYT (BFI2000-1360), FIS (01-0948, PI021517) and ISCIII (red CIEN, nodo Fundación Carlos Haya)     

An immunocytochemical and ultrastructural study of a specialized glial region of the medulla oblongata of the adult and juvenile grey mullet

Electron microscopy together with glial fibrillary acidic protein (GFAP) and vimentin immunocytochemistry reveals the presence of a specialized glial region in the octavolateral area of the medulla oblongata of adult and juvenile grey mullets, Chelon labrosus. Glial cells, which can he characterized as ependymal and subependymal cells, originate in the walls of the lateral recess of the fourth ventricle. They contain numerous gliofilaments and are connected through frequent gap junctions. Electron microscopy also reveals the lack of nerve cell parikarya and processes in this region. Immunocytochemistry reveals that the glial cells are strongly GFAP-positive in both adults and juveniles, whereas vimentin is only detected in this region in juveniles. The meninges associated with this glial region contains connective fibres, formed of very thick collagen fibres, that arc metachromatic and PAS-positive under light microscopy. These findings strongly support a structural role for this medullary specialization. Differences between adults and juveniles in the distribution of GFAP- and vimentin-immunoreactive structures in other regions of the medulla oblongata are also reported.     

Experimental protein malnutrition as a causative factor in the histological and histochemical disruption of the ependymal cells of the third ventricla and cervical central canal of squirrel monkeys.

The histologic disturbances of the cuboidal and columnar ependymal cells of the neonates under the extrinsic influence of maternal protein deprivation during most of the gestation period have been demonstrated in the central canal of the cervical spinal cord and III ventricle of the squirrel monkey brain. The control animals whose mothers were maintained on high protein diets showed an unbroken ependymal layer with an intact glial fiber layer and the subependymal cell plate. Taking birth-weight as an indication of the degree of malnutrition, maximum disruption of the ependymal layer was observed in those animals born around 80 g birth-weight. In the latter, most of the ependymal cells lost their characteristic arrangement and columnar shapes. The ependymal cells of the manourished animals show a marked reduction of oxidative enzyme content and relatively active glycogen metabolism as compared to the healthy controls. The significance of these changes has been discussed in the light of parallels in the embryological development and, to some extent, functions between the orinary neurons, ependymal cells of the choroid plexus, other ependymal cells and the glial cells. In this context, the changes observed in the present studies under the effect of protein malnutrition must be taken as part of overall changes experienced by various kinds of cells comprising the nervous system.     

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     

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.     

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.     

Immunohistochemical localization of basic fibroblast growth factor in ependymal cells of the rat lateral and third ventricles.

The presence of basic fibroblast growth factor (bFGF) in the third and the lateral ventricular ependyma of the rat has been investigated under the light microscope using a polyclonal antibody against bFGF, through the unlabelled peroxidase-antiperoxidase procedure; bFGF-immunoreactivity (bFGF-IR) is observed in the entire ependymal cell layer of ventricles. Also ependymal tanycytes within the inferior portion of the wall and floor of the third ventricle show bFGF-IR. Tanycytic processes are in close contact with hypothalamic capillaries. The present study strongly suggests that brain ependymal bFGF plays unidentified roles unrelated to its angiogenic or mitogenic capacities.     

Ependymal specializations

Summary The primary plexus of the toad hypothalamic-adenohypophysial portal system has two types of loops. The short loops are localized in the external region of the median eminence and surrounded by nerve endings and glial cells. The long loops approach the ependymal lining of the median eminence. The ascending and descending branches of these loops are surrounded by nerve and ependymal endings and glial cells. The actual subependymal portion of the long loops is virtually in contact with ependymal processes only, which form a cuff interposed between this portion of the long loops and the fibres of the hypothalamic-neurohypophysial tract. Many of the vascular endings of the ependymal processes have electron dense granules whose diameter ranges between 700 and 1400 Å. The ultrastructure of the ependymal cells suggests that these granules are transport material and not secretory material.This anatomical arrangement linking the ependyma of the median eminence and the long loops of the primary plexus of the hypothalamic-adenohypophysial portal system makes the possibility of an interrelationship between the cerebrospinal fluid and the portal blood very considerable.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina. The author takes great pleasure in thanking Prof. H. Heller for his constant interest and criticism.     

Cellules riches en glycogène dans l'épendyme du ventricule latéral du cerveau de rat

Summary The ependyma of the lateral ventricle of the rat brain was investigated at different ages from 20 days to adulthood.A particular cell type occurs in the external wall of the ventricle, where the proliferative subependymal layer is present. These cells found at all ages studied, are characterized by a high content of glycogen, and a structure different of typical ependymal cells.A large number of nerve endings is situated in close vicinity of these cells, either free in the ventricle lumen, or sometimes ensheathed in the cells. No synapse was found between these endings an the glycogen-rich cells.These glycogen-rich cells undergo several modifications with age: their glycogen content is reduced in the adult, and they acquire a few cilia and gliofilaments. It is suggested that they represent a transitory differentiation of the ependyma, functionally linked with the proliferative subependymal layer.

     

Golgi-like immunostaining of pituicytes and tanycytes positive for glial fibrillary acidic protein in the neurohypophysis of the Mongolian gerbil (Meriones unguiculatus)

Summary Glial cells that contain the glial fibrillary acidic protein (GFAP; the major protein constituent of glial filaments) were stained immunohistochemically in thick frozen sections of the neurohypophysis of the Mongolian gerbil (Meriones unguiculatus). The resulting Golgi-like images provided informations on cytological features and distributional patterns of tanycytes and pituicytes. In the proximal median eminence, numerous bundled processes of tanycytes were revealed. They emerged from the ependymal and subependymal layer and mostly reached the brain surface. Several tanycytic processes extended into the anatomical neural stalk. In the whole neural lobe, a dense network of GFAP-immunoreactive pituicyte processes was visualized. Stained pituicytes were highly asymmetric and exhibited a great morphological variability. Immunopositive fibers which were encountered in the intermediate lobe might be derived from pituicytes. Electron-microscopically further evidence was obtained that GFAP-positive pituicytes correspond to filament-rich fibrous pituicytes at the ultrastructural level.     

Neurogenesis in the Adult Mammalian Brain

The concept of the CNS cell composition stability has recently undergone significant changes. It was earlier believed that neurogenesis in the mammalian CNS took place only during embryonic and early postnatal development. New approaches make it possible to prove that neurogenesis takes part even in the adult brain. The present review summarizes the data about the neural stem cell. It has been demonstrated that new neurons are constantly formed in adult mammals, including man. In two brain zones, subventricular zone and dentate gyrus, neurogenesis appears to proceed throughout the entire life of mammals, including man. The newly arising neurons are essential for some important processes, such as memory and learning. Stem cells were found in the subependymal and/or ependymal layer. They express nestin and have a low mitotic activity. During embryogenesis, the stem cell divides asymmetrically: one daughter cell resides as the stem cell in the ependymal layer and another migrates to the subventricular zone. There it gives rise to a pool of dividing precursors, from which neural and glial cells differentiate and migrate to the sites of final localization. The epidermal and fibroblast growth factors act as mitogens for the neural stem cell. The neural stem cell gives rise to the cells of all germ layers in vitro and has a wide potential for differentiation in the adult organism. Hence, it can be used as a source of various cell types of the nervous tissue necessary for cellular transplantation therapy.     

Neurogenesis in the adult mammalian brain

The concept of the CNS cell composition stability has recently undergone significant changes. It was earlier believed that neurogenesis in the mammalian CNS took place only during embryonic and early postnatal development. New approaches make it possible to obtain new results overriding the dogma that neurogenesis is impossible in the adult brain. The present review summarizes the information about the neural stem cell. It has been demonstrated that new neurons are constantly formed in adult mammals, including man. In two brain zones, subventricular zone and denate gyrus, neurogenesis appears proceed throughout the entire life of mammals, including man. The newly arising neurons are essential for some important processes, such as memory and learning. Stem cells were found in the subependymal and/or ependymal layer. They express nestin, and have a low mitotic activity. During embryogenesis, the stem cell divides asymmetrically: one daughter cell resides as the stem cell in the ependymal layer and another migrates to the subventricular zone. There it gives rise very fast to a pool of dividing precursors, from which neural and glial cells differentiate and migrate to the sites of final localization. The epidermal and fibroblast growth factors act as mitogens for the neural stem cell. The neural stem cell gives rise to the cells of all germ layers in vitro and has a wide potential for differentiation in the adult organism. Hence, it can be used as a source of various cell types of the nervous tissue necessary for cellular transplantation therapy.     

The subcommissural organ of the frog Rana perezi is innervated by nerve fibres containing GABA

The innervation of the frog subcommissural organ was studied by light-microscopic and ultrastructural immunocytochemistry using antisera against serotonin, noradrenaline, dopamine, gamma-aminobutyric acid (GABA), glutamic acid decarboxylase, different GABA receptor subunits and bovine Reissner's fibre material (AFRU). In the proximity of the organ, serotonin- and noradrenaline-containing fibres were rare whereas dopamine-immunoreactive fibres were more numerous. Many GABA- and glutamic acid decarboxylase-containing nerve fibres were found at the basal portion of the ependymal cells of the subcommissural organ. Under the electron microscope, these GABA-immunolabelled nerve endings appeared to establish axoglandular synapses with secretory ependymal cells of the subcommissural organ. In addition, the secretory ependymal cells expressed high amounts of the beta2-subunit of the GABA(A) receptor. Since GABA-immunoreactive neurons were present in the frog pineal organ proper and apparently contributed axons to the pineal tract, we suggest that at least part of the GABAergic fibres innervating the frog subcommissural organ could originate from the pineal organ.     

A Golgi study on the cerebrospinal fluid (CSF)-contacting neurons in the paraventricular nucleus of the Pekin duck

The present investigation based on classical neurohistological techniques (Nissl-staining, Golgi-impregnation) was focussed on the cytoarchitecture of the periventricular layer of the paraventricular nucleus in the Pekin duck. This region is endowed with intraependymal neurons, the perikarya of which are mostly covered by a thin ependymal lamella. Several of the intraependymal neurons were shown to give rise to dendrites extending into the third ventricle. An additional population of nerve cells located in the deeper layers of the periventricular region also gained direct access to the cerebrospinal fluid by means of long dendrites terminating with a bulbous-like swelling in the third ventricle. This cerebrospinal fluid (CSF)-contacting dendrite branched off several times in a rectangular fashion to give rise to collaterals running in the subependymal or periventricular layers. The axons of these CSF-contacting neurons were followed into the magnocellular portion of the paraventricular nucleus. Small bipolar nerve cells with processes parallel to the surface of the third ventricle occupied a subependymal position. The isodendritic magnocellular neurons of the paraventricular nucleus emitted dendritic processes that reached the basal pole of the ependymal cells. The complex arrangement of the periventricular layer of the paraventricular nucleus might provide the structural basis for the mechanisms of cerebral osmoreception defined by means of physiological parameters.     

Aminoacyl-histidine dipeptides in the glial cells of the adult rabbit forebrain

The mammalian nervous system contains high amounts of the aminoacyl-histidine dipeptides carnosine and homocarnosine. In the brain, they prevalently occur mainly in glial and ependymal cells, their role(s) still remaining obscure. In vitro studies indicate that these molecules exert diverse protective effects, and in vivo they are frequently associated with extracellular fluid compartments. Recently, carnosine-like immunoreactivity has been found in the subependymal layer (SEL) of adult rodents, a region endowed with persistent cell proliferation and migration. Unlike rodents, the SEL of the rabbit has a persistent olfactory ventricle. We show here that the morphologic organization of the SEL is different in these species, with particular reference to the glial/non glial cell compartments. The distribution of carnosine-like immunoreactivity in the rabbit displays some differences only within the SEL, which could be linked to its arrangement and compartmentalization.     

Glial fibrillary acidic protein-like immunoreactive ependymal elements in the third ventricle of the rat. A study at different stages of development.

Using the peroxidase-antiperoxidase method a study was made of the cells immunoreactive to glial fibrillary acidic protein (GFAP) anti-serum in the ependyma of the third ventricle of the rat at different stages of growth. Most of the ependymal cells of the third ventricle were seen to be unreactive to this protein; however, it was sometimes possible to observe some GFAP-immunoreactive ependymocytes and occasionally other immunoreactive cellular types, such as tanycytes and supraependymal cells. Despite this, the most frequent localization of the elements immunoreactive to the protein adopted the shape of an immunoreactive subependymal band situated parallel to the ventricular wall. As the weights of the animals increased an increase in the elements immunoreactive to this protein could be observed in all the zones considered, there being no differences between the male and female animals.     

Glial fibrillary acidic protein and vimentin immunoreactivity of astroglial cells in the central nervous system of the African lungfish, Protopterus annectens (Dipnoi: Lepidosirenidae)

The distribution of glial intermediate filament molecular markers, glial fibrillary acidic protein (GFAP), and vimentin, in the brain and spinal cord of the African lungfish, Protopterus annectens, was examined by light microscopy immunoperoxidase cytochemistry. Glial fibrillary acidic protein immunoreactivity is clear and is evident in a radial glial system. It consists of fibers of different lengths and thicknesses that are arranged in a regular radial pattern throughout the central nervous system (CNS). They emerge from generally immunopositive radial ependymoglia (tanycytes), lining the ventricular surface, and are directed from the ventricular wall to the meningeal surface. These fibers give rise to endfeet that are apposed to the subpial surface and to blood vessel walls forming the glia limitans externa and the perivascular glial layer, respectively. GFAP-immunopositive star-shaped astrocytes were not found in P. annectens CNS. In the gray matter of the spinal cord, cell bodies of immunopositive radial glia are displaced from the ependymal layer. Vimentin-immunopositive structures are represented by thin fibers mostly localized in the peripheral zones of the brain and the spinal cord. While a few stained fibers appear in the gray matter, the ependymal layer shows no antivimentin immunostaining. In P. annectens the immunocytochemical response of the astroglial intermediate filaments is typical of a mature astroglia cell lineage, since they primarily express GFAP immunoreactivity. This immunocytochemical study shows that the glial pattern of the African lungfish resembles that found in tetrapods such as urodeles and reptiles. The glial pattern of lungfishes is comparable to that of urodeles and reptiles but is not as complex as that of teleosts, birds, and mammals.     

An immunofluorescence microscopical study of the neurofilament triplet proteins, vimentin and glial fibrillary acidic protein within the adult rat brain

A collection of antibodies specific to different intermediate filament proteins were applied to frozen sections of adult rat brains. The relative distribution of these proteins was then studied using double label immunofluorescence microscopy. Antibodies specific to each of the neurofilament "triplet" proteins (of approximate molecular weight 68 K, 145 K and 200 K) stained exclusively neuronal structures. The distribution of these three antigens was in general identical, except that certain neurofilament populations such as those in the dendrites and cell bodies of pyramidal cells of the hippocampus and cerebral cortex, contained relatively little if any 200 K protein. Some neurone populations, such as the granule cells of the cerebellar cortex, could not be visualized by neurofilament antibodies, indicating that neurofilaments may not be essential for function of all neurones in vitro. Antibodies to GFA and vimentin stained an entirely different population of processes, none of which stained with any of the neurofilament antibodies. Vimentin antibody stained sheath material around the brain, a monolayer of ependymal cell bodies lining the ventricles, fibrous material associated within the choroid plexus, the walls of blood vessels and capillaries, and the processes of cells in certain regions. GFA antibody stained a second layer of sheath material under the vimentin layer, and numerous processes visible throughout the brain. Some specific populations of GFA-positive processes proved to stain also with vimentin. These included the processes of Golgi "epithelial" cells (Bergmann glial fibres), those of certain astrocytes in bundles of myelinated fibers. In addition, some processes apparently derived from ependymal cells proved to stain for both vimentin and GFA, whilst other could only be reliably visualized by vimentin alone. These results are discussed in terms of the previously described morphological characteristics of the various cell types of the brain.     

The neurohypophysis of the crested newt

Summary In the median eminence of the newt a medial region and two lateral regions are described.In cross section, the medial region appears to be made up of 1) an outer or glandular zone (Zone I) containing aldehyde-thionine-positive and negative nerve fibres and blood capillaries. Nerve fibres appear aligned in palisade array along the capillaries. 2) An inner zone (Zone II) made up of a) a layer of aldehyde-thionine-positive nerve fibres (fibrous layer) belonging to the preoptic hypophyseal tract and b) a layer of ependymal cells lining the infundibular lumen and reaching the blood vessels with their long processes.The lateral regions display a less pronounced stratification and aldehyde-thionine positive nerve fibres are nearly absent.A slender lamina (ependymal border) containing mainly aldehyde-thionine-positive nerve fibres and ependymal cells connects the median eminence to the pars nervosa.At the ultrastructural level, in the outer zone of the medial region at least 4 types of nerve fibres and nerve endings are identified:Type I nerve fibres containing granular vesicles of 700–1000 Å and clear vesicles (250–400 Å).Type II nerve fibres containing granular vesicles and polymorphous granules of 900–1300 Å and clear vesicles (250–400 Å).Type III nerve fibres containing dense granules of 1200–2000 Å and clear vesicles of 250–400 Å.Type IV nerve fibres containing only clear vesicles of 250–400 Å. In the inner zone too, all these nerve fiber types are found among ependymal cells, while the fibrous layer consists of nerve fibres containing granules of 1200–2000 Å in diameter.In the lateral regions Type I, Type II and Type IV nerve fibres and their respective perivascular terminals are found; axons containing dense granules (1200–2000 Å) are scanty. In these regions typical synapses between Type I nerve fibres and processes rich in microtubules are visible.The classification and functional significance of nerve fibres in the median eminence are still unsolved, but it may be assumed that nerve fibres of the medial region belong to both the preoptic hypophyseal and tubero hypophyseal tract, while the lateral regions are characterized by nerve fibres of the tubero hypophyseal tract. Peculiar specializations of the ependymal cells in the median eminence of the newt are also discussed.Work supported by a grant from the Consiglio Nazionale delle Ricerche.The authors are indebted to Mr. G. Gendusa and P. Balbi for technical assistance.