Immunocytochemical demonstration of retinal S-antigen in the pineal organ of four mammalian species

By means of immunocytochemistry retinal S-antigen is selectively demonstrated in retinal photoreceptor cells of the rat and in pinealocytes of the hedgehog, rat, gerbil and cat. Brain areas surrounding the pineal organ are immunonegative. The immunoreactive material is evenly distributed in the perikarya of the cells. Occasionally, inner segments of retinal photoreceptors and processes of pinealocytes are also stained. The outer segments of retinal photoreceptors display a strong immunoreaction. In both pinealocytes and retinal photoreceptors the intensity of the immunoreaction varied considerably among individual cells. The immunocytochemical demonstration of retinal S-antigen in mammalian pinealocytes indicates that these cells still bear characteristics of photoreceptors. This finding is in accord with the concept that mammalian pinealocytes are derived from pineal photoreceptor cells of poikilothermic vertebrates.     

Rod-opsin immunoreaction in the pineal organ of the pigmented mouse does not indicate the presence of a functional photopigment

The aim of the present study was to characterize the rod-opsin immunoreaction in the mammalian pineal organ. Pigmented mice (strain C57BL) were selected as the animal model. Immunocytochemical investigations involving the use of highly specific polyclonal and monoclonal antibodies against bovine rod-opsin (the apoprotein of the photopigment rhodopsin) showed that approximately 25% of all pinealocytes were rod-opsin immunoreactive. Immunoblotting techniques revealed three protein bands of approximately 40, 75, and 110 kDa; these were detected by the monoclonal antibody and the polyclonal antiserum in retinal and pineal extracts. These protein bands presumably represented the monomeric, dimeric and trimeric forms of rod-opsin. The amount of rod-opsin in retina and pineal organ was quantified by means of an enzyme-linked immunosorbent assay. This yielded 570±30 pmoles rod-opsin per eye and 0.3±0.05 pmoles rod-opsin per pineal organ. High pressure liquid chromatography analysis of whole eye extracts demonstrated the chromophoric group of the photopigment rhodopsin, 11-cis retinal, and its isomer, all-trans-retinal. A shift from 11-cis retinal to all-trans-retinal was found upon light adaptation. No retinals were detected in the pineal organ. Autoradiographic investigations showed that ³H-retinol, intraperitoneally injected into the animals, was incorporated into the outer and inner segments of retinal photoreceptors, but not into the pineal organ. It is concluded that the mouse pineal organ contains the authentic apoprotein of rhodopsin but that it lacks retinal derivatives as essential components of all known vertebrate photopigments. Consequently, the photoreceptor-specific proteins of the mammalian pineal organ are not involved in photoreception and phototransduction, but may serve other functions to be explored in future studies.     

Ultrastructure of serotonin-containing cells in the pineal organ of Lampetra planeri (Petromyzontidae)

Summary The ultrastructure of the cells containing residual bodies (Collin, 1969) was investigated in the pineal organ of Lampetra planeri. These cells are characterized by their indoleamine metabolism (Meiniel, 1978; Meiniel and Hartwig, 1980). Morphologically, they belong mainly to two types: (1) a photoreceptor cell type, and (2) a pinealocyte cell type. The first type is present in the pineal sensory epithelium and in the atrium, while the second is observed in the deep part of the atrium. Intermediate cell types are rare. All these cells are characterized by the presence of voluminous dense bodies, the 5-HT-storing structures, in their cytoplasm.The elongated cone-type photoreceptor cells show a segmental organization and well-developed outer segments consisting of short disks (2–3 m), while their basal pedicles form synapses with the dendritic processes of neurons. The pinealocytes are spherical or oval in shape, their receptor poles being regressed to cilia of the 9+0 type. In these cells, no synaptic ribbons have to date been observed. In both cell types a Golgi apparatus is present producing dense granules 130 nm in diameter and a polymorphous dense material.The photoreceptor cells most probably respond to light and transmit a sensory (i.e., nervous) message. In addition, they produce and metabolize indoleamines, probably including, melatonin (Meiniel, 1978; Meiniel and Hartwig, 1980). The pinealocytes, in spite of their loss of direct photosensitivity, retain their capacity to metabolize indoleamines (Meiniel, 1978; Meiniel and Hartwig, 1980).The presence, in the same pineal organ, of another photoreceptor cell type (cf. Collin, 1969–1971) differing morphologically as well as biochemically (no detectable indoleamine metabolism) from the photoreceptor cell type described in the present investigation, points to the existence of two different sensory cell lines: (1) a pure photoreceptor line, and (2) a photoneuroendocrine line. The phylogenetic evolution of these two cell lines is discussed in terms of functional analogy.     

The parapineal and pineal organs of the elver (glass eel), Anguilla anguilla L.

Summary The parapineal organ of the glass eel (elver) consists of approximately 400 cells and is situated to the left of the connection of the pineal stalk to the third ventricle. A conspicuous nerve tract containing approximately 350 fibers arises from the parapineal organ and runs in spatial relationship to the habenular commissure toward the left habenular nucleus. The dominating cell type of the parapineal organ of the elver is a neuron (sensory neuron) of small diameter provided with atypical cilia (9×2+0, or rarely 8×2+0 types). Well-developed photoreceptor outer segments are lacking, and no interstitial cells of ependymal type have been observed with certainty in the parapineal organ. The axonal processes from the nerve cells form the tract leaving the parapineal organ.The pineal organ proper of the elver consists of photoreceptor cells with well-developed outer segments, interstitial cells of ependymal type, and ganglion cells. Axons from the latter form the pineal tract, which leaves the pineal organ and runs in close contact with the subcommissural organ toward the posterior commissure. The proximal part of the pineal stalk contains only a few photoreceptor cells the outer segments of which are less developed than those of the pineal body and the distal part of the pineal stalk.     

Immunocytochemical and electron-microscopic investigations of the pineal organ in adult agamid lizards,Uromastix hardwicki

Summary Lacertilian species display a remarkable diversity in the organization of the neural apparatus of their pineal organ (epiphysis cerebri). The occurrence of immunoreactive S-antigen and opsin was investigated in the retina and pineal organ of adult lizards, Uromastix hardwicki. In this species, numerous retinal photoreceptors displayed S-antigen-like immunoreactivity, whereas only very few pinealocytes were labeled. Immunoreactive opsin was found neither in retinal photoreceptors nor in pinealocytes. Electron microscopy showed that all pinealocytes of Uromastix hardwicki resemble modified pineal photoreceptors. A peculiar observation is the existence of a previously undescribed membrane system in the inner segments of these cells. It is evidently derived from the rough endoplasmic reticulum but consists of smooth membranes. The modified pineal photoreceptor cells of Uromastix hardwicki were never seen to establish synaptic contacts with somata or dendrites of intrapineal neurons, which are extremely rare. Vesiclecrowned ribbons are prominent in the basal processes of the receptor cells, facing the basal lamina or establishing receptor-receptor and receptor-interstitial type synaptoid contacts. Dense-core granules (60–250 nm in diameter) speak in favor of a secretory activity of the pinealocytes. Attention is drawn to the existence of receptor-receptor and receptor-interstitial cell contacts indicating intramural cellular relationships that deserve further study.Supported by the Deutsche Forschungsgemeinschaft (Ko 758/31) and the Deutscher Akademischer Austauschdienst (Senior DAAD Research Fellowship to M.A.H.)     

The role of the pineal and the retinae in the expression of circadian locomotor rhythmicity in the ruin lizard,Podarcis sicula

Summary A marked interspecific variability in the role played by the pineal and the retinae characterizes the circadian system of lizards. I examined the role played by these structures in a new model species, the ruin lizard, Podarcis sicula. In constant temperature and darkness pinealectomy as well as bilateral removal of the retinae produced significant changes (both lengthening and shortening) in the freerunning period of locomotor rhythms. Circadian activity time was also affected by pinealectomy. Circadian locomotor rhythmicity persisted in all cases even when both operations were combined in the same individuals. This demonstrates in Podarcis sicula the existence of an oscillatory system outside the pineal and the retinae which can drive locomotor rhythms. The period changes recorded after pinealectomy as well as after bilateral removal of the retinae specifically suggest that both the pineal and the retinae play a modulating role on circadian oscillators located elsewhere in the system, with the final effect of stabilizing the overt rhythms.Abbreviations DD constant darkness - LL constant light - PIN-X pinealectomy - RET-X bilateral removal of the retinae - SHAM sham pinealectomy - circadian activity time - freerunning circadian period     

Structural and functional differentiation of the embryonic chick pineal organ in vivo and in vitro

Summary The development of sensory structures in the pineal organ of the chick was examined by means of scanning electron microscopy from embryonic day 10 through day 12 post-hatching. At embryonic day 10, the wall of the tubules within the pineal primordium is composed of cells with unspecialized luminal surface. Differentiation of sensory structures starts at embryonic day 12 when pinealocytes and supporting cells can be distinguished. Pinealocytes are recognized by virtue of an inner segment only rarely endowed with a cilium, whereas supporting cells exhibit numerous short microvilli. Further differentiation of the sensory apparatus is achieved by development of an oval-shaped, biconcave swelling at the tip of the cilium, 1×2 m in size, and a collar of long microvilli at the base of the inner segment. Membrane specializations of sensory cilia, however, were not detected. Since during embryonic life new tubules and follicles are continuously formed, all stages of differentiation of sensory structures are found in the chick pineal organ during the second half of the incubation period and the first two weeks after hatching. In 200-m-thick Vibratome sections of chick-embryo pineal organs cultured in medium BM 86 Wissler for periods up to 13 days the cytodifferentiation parallels the development in vivo. Using an organ-culture system the 24-h release of melatonin into the culture medium was measured by means of radioimmunoassay after solid-phase extraction. At embryonic day 10, the 24-h secretion of melatonin was at the lower range of detection of the RIA (5 pg). The rapid increase in 24-h secretion in melatonin until hatching (50 g) is approximated by an exponential curve.Preliminary results of this study were reported at the Versammlung der Anatomischen Gesellschaft in Lübeck, 1986 (Möller 1987). Supported by the Deutsche Forschungsgemeinschaft (MO234/9-2)     

A pineal ganglion associated with the pineal tract in the domestic fowl

Summary A ganglion-like aggregate consisting of acetyl-cholinesterase-positive neurons was demonstrated in the pineal organ of the domestic fowl by means of light and electron microscopy. This ganglion is located in juxtaposition with the pineal tract at the posterior (caudal) aspect of the pineal stalk. Numerous large and small neurons formed the ganglion in 40-day-old domestic fowl. Some of these nerve cells established direct neuro-neuronal contacts, others were surrounded by satellite cells. These ganglion cells displayed axo-somatic and axo-dendritic synapses. The above-mentioned cluster of nerve cells may be considered as a pineal ganglion. Its central or peripheral nature is open to discussion. Send offprint requests to: Dr. K. Wake, Department of Anatomy, Faculty of Medicine, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, 113, Japan     

The pineal gland of the gerbil,Meriones unguiculatus

Summary Electron microscopy was employed in a study of the pineal gland of the Mongolian gerbil (Meriones unguiculatus). It was determined that the gerbil pineal gland contains pinealocytes and glial cells with the pinealocytes being the predominant cell type. The pinealocytes contain numerous organelles traditionally considered as being either synthetic or secretory in function such as an extensive Golgi region, smooth (SER) and rough (RER) endoplasmic reticulum, secretory vesicles and microtubules. Other cytoplasmic components are also present in the pinealocytes (synaptic ribbons, subsurface cisternae) for which no function has been assigned. Dense-cored vesicles are rare. Vacuolated pinealocytes are present and appear to be intimately associated with the formation of the pineal concertions. Evidence presented supports the proposal that the concretions form within the vacuoles. Once the concretions reach an enlarged state, the vacuolated pinealocytes break down and the concretions are thus extruded into the extracellular space where they apparently continue to increase in size. The morphology of the glial cells was interpreted as indicative of a high synthetic activity. The glial cells contain predominantly the rough variety of endoplasmic reticulum and form an expansion around the wide perivascular area.Supported by NSF grant PCM 77-05734     

The pineal gland of the gerbil,Meriones unguiculatus

Summary By means of morphometric analytical procedures, a diurnal rhythm in the cellular volume of gerbil pinealocytes was determined. This rhythm has been attributed primarily to a change in the cytoplasmic volume of the pinealocytes which is low during the daylight hours and increases to reach a peak during the middle of the dark period. At the ultrastructural level, six cytoplasmic components of the pinealocytes were found to exhibit a rhythm: free cytoplasm, smooth endoplasmic reticulum (SER), rough endoplasmic reticulum (RER) and ribosomes, secretory vesicles, microtubules, and mitochondria. The presumptive secretory vesicles and the microtubules reached a peak in volume one hour before lights-off. It is suggested that lights-on and lights-off both signal a decrease in size and/or number of the secretory vesicles. The SER and RER/ribosomes reached their peak volume one hour after lights-off which is interpreted as indicating a peak in indoleamine synthesis and protein synthesis, respectively. The volume of free cytoplasm exhibits two peaks; one occurs one hour before lights-off while the second peak occurs in the middle of the dark phase. It is suggested that, although part of the secretory product of the pinealocyte may be present in dense-cored vesicles, other locations could include the free cytoplasm and clear secretory vesicles.Supported by NSF grant #PCM 77-05734     

Interstitial and parenchymal cells in the pineal gland of the golden hamster

Summary A combined thin-section/freeze-fracture study was performed on the superficial pineal gland of the golden hamster, comparing the parenchymal and interstitial cells of this animal with those previously investigated in rats. In contrast to rats, no gap junctions and gap/tight junction combinations could be found between pineal parenchymal cells of the hamster. Furthermore, the interstitial cells of the hamster pineal gland were found to have large flat cytoplasmic processes, which abut over large areas equipped with tight junctions. In thin sections, profiles of interstitial cell processes were seen to surround groups of pinealocytes. Interstitial cells and their sheet-like, tight junction-sealed processes thus appear to delimit lobule-like compartments of the hamster pineal gland. Because the classification of the interstitial cells is uncertain, the expression of several markers characteristic of mature and immature astrocytes and astrocyte subpopulations has been investigated by indirect immunohistology. Many of the non-neuronal elements in the pineal gland are vimentin-positive glial cells, subpopulations of which express glial fibrillary acidic protein (GFA) and C1 antigen. The astroglial character of these cells is supported by the lack of expression of markers for neuronal, meningeal and endothelial cells. M1 antigen-positive cells have not been detected.Supported by a grant from Deutsche Forschungsgemeinschaft (Scha 185/9-2)     

Ultrastructural visualization of exocytosis in the pig pineal gland

Summary The pinealocytes of the pig contain conspicuous dense bodies, the nature and role of which are not yet fully elucidated. The aim of this study was to demonstrate whether or not these structures are involved in the secretion process. The tannic acid-Ringer incubation (TARI)-method, which allows a clear-cut ultrastructural study of secretory discharge by exocytosis, has been used. The results indicate that pig pinealocytes release the content of the dense bodies with an amorphous inner structure into the extracellular space via exocytosis and that this secretion is quantitatively important. The secreted material is proteinaceous in nature; this indicates that polypeptides are released by the pineal.     

Granulated vesicles in the pineal gland of the mouse

Summary Pineal glands of normal adult mice, 7 to 42 days after bilateral superior cervical gangliectomy and 5 and 16 hours after one dose of reserpine (10 mg/kg) were studied under the electron microscope. The architecture of the gland is basically similar to that of other mammalian pineal glands previously studied. Mouse pinealocytes are polymorphic cells with perivascular and intercellular processes. Its most prominent feature is the presence of abundant granulated vesicles with a mean diameter of 1100 Å and a dense core of about 800 Å intermingled with clear vesicles of similar size distributed throughout the cytoplasm and more concentrated in perivascular and intercellular processes. These processes were seen in continuity with the perykaryon and remained after bilateral superior cervical gangliectomy. Processes containing the plurivesicular component of adrenergic nerves situated in perivascular and intercellular spaces disappeared after bilateral superior cervical gangliectomy. Reserpine depleted small granulated vesicles of nerves but the larger ones of perikaryon and polar processes remained almost unchanged.The histochemical technique of Wood was positive for catechol- and indolamines in the nerves. The reaction was negative in the perikaryon and polar processes.The significance of these findings is discussed.This work has been supported by grants of the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina and U.S. Air Force AF-AFOSR 67-0963 A.I am greatly indebted to Miss Haydee Agoff and Mr. Alberto Saenz for their skillful technical assistance.     

Regressive post-hatching development of acetylcholinesterase-positive neurons in the pineal organs of Coturnix coturnix japonica and Gallus gallus

Summary Distribution and number of acetylcholinesterase-positive neurons were studied in the Japanese quail and the domestic fowl during the post-hatching period by means of the acetylcholinesterase method. For comparison, the development of the catecholamine-containing (sympathetic) pinealopetal fibers of the domestic fowl was demonstrated with the use of the glyoxylic acid method. The number of acetylcholinesterase-positive ganglion cells in the pineal organs of both avian species decreased rapidly after hatching, with a concentration of these elements in the basal portion (stalk) of the pineal organ.In 3-day-old chickens, perivascular catecholamine-containing nerve fibers penetrate the antero-lateral walls of the pineal organ and are found exclusively in the interfollicular and perivascular tissues. In 13-day-old and adult fowl, these fibers increase in number and terminate not only in the interfollicular space but also in the neuroepithelial parenchyma of the pineal body.The ontogenetic regression of the sensory structures paralleled by an expanding sympathetic innervation in the pineal organ of a galliform species resembles somewhat the process of phylogenetic transformation leading from pineal sense organs to pineal glands.This work was supported by a grant (No. 56480080) from the Ministry of Education, Science and Culture of Japan.Fellow of the Alexander von Humboldt Foundation (1982).     

The influence of noradrenaline on the process of protein/peptide secretion in the mammalian pineal organ

Summary From studies conducted with the pineal organ of the mouse, it was ascertained that for the in vitro investigation of secretory processes (synthesis and release) of proteic/peptidic compound(s), a culture time of 5 to 14 days is optimal. A 5-day organ culture was therefore chosen to study the effects of noradrenaline on these secretory processes.Addition of noradrenaline to the culture medium provokes, in pineal explants of the normal mouse and the eyeless mouse, an inhibition of the secretory process, characterized by the formation of granular vesicles. In the hamster and rat, however, opposite results were obtained. Moreover, it appears that noradrenaline, at least in the rat, may also be involved in the regulation of the ependymal-like secretory process.The present results indicate clearly that noradrenaline (thus, the sympathetic innervation) is implicated in the regulation of the production of proteic/peptidic hormonal agents, but that the effect of this neurotransmitter is species-specific. This could explain the numerous contradictory results reported in the literature.IBRO/UNESCO fellow     

Immunocytochemical localization of serotonin and photoreceptor-specific proteins (rod-opsin,S-antigen) in the pineal complex of the river lamprey,Lampetra japonica,with special reference to photoneuroendocrine cells

Summary The pineal complex of the river lamprey, Lampetra japonica, was examined by means of immunocytochemistry with antisera against serotonin, the precursor of melatonin, and two photoreceptor proteins, rod-opsin (the apoprotein of the photopigment rhodopsin) and S-antigen. Serotonin-immunoreactive cells were observed in both the pineal and the parapineal organ. The proximal portion of the pineal organ (atrium) comprised numerous serotonin-immunoreactive cells displaying spherical somata. In the distal end-vesicle of the pineal organ, the serotonin-immunoreactive elements resembled photoreceptors in their size and shape. These cells projecting into the pineal lumen and toward the basal lamina were especially conspicuous in the ventral portion of the end-vesicle. In addition, single serotonin-immunoreactive nerve cells were found in this location. Retinal photoreceptors were never seen to contain immunoreactive serotonin; amacrine cells were the only retinal elements exhibiting serotonin immunoreaction. Strong S-antigen immunoreactivity was found in numerous photoreceptors located in the pineal end-vesicle. In contrast, the S-antigen immunoreactivity was weak in the spherical cells of the atrium. Thus, the pattern of S-antigen immunoreactivity was roughly opposite to that of serotonin. Similar findings were obtained in the parapineal organ. The rod-opsin immunoreaction was restricted to the outer segments of photoreceptors in the pineal end-vesicle and parapineal organ. No rodopsin immunoreactive outer segments occurred in the proximal portion of the atrium. Double immunostaining was employed to investigate whether immunoreactive opsin and serotonin are colocalized in one and the same cell. This approach revealed that (i) most of the rodopsin-immunoreactive outer segments in the end-vesicle belonged to serotonin-immunonegative photoreceptors; (ii) nearly all serotonin-immunoreactive cells in the end-vesicle bore short rod-opsin-immunoreactive outer segments protruding into the pineal lumen; and (iii) the spherical serotonin-immunoreactive cells in the pineal stalk lacked rod-opsin immunoreaction and an outer segment. These results support the concept that multiple cell lines of the photoreceptor type exist in the pineal complex at an early evolutionary stage.     

Vascular permeability (problem of the blood-brain barrier) in the pineal organ of the rainbow trout,Salmo gairdneri

Summary The problem of the blood-brain barrier in the pineal organ of the rainbow trout, Salmo gairdneri, was investigated following intraperitoneal or intracardial injections of several tracers and dyes with different molecular weights. As demonstrated at the light-microscopic level, repeated injections of trypan blue or horseradish peroxidase (HRP) resulted in an accumulation of these substances in the pineal epithelium (parenchyma). By use of the electron microscope, HRP was found in electron-dense bodies, probably lysosomes, in (i) the endothelial cells and perivascular macrophages 4 h after intraperitoneal injection, (ii) the supporting cells and intrapineal or luminal macrophages 8 h after injection, and (iii) the receptor cells 24 h after injection of the tracer. Ferritin particles penetrated the fenestrated endothelium of pineal capillaries. They were confined to vesicles, vacuoles and the smooth endoplasmic reticulum of the supporting cells as well as to the synaptic vesicles and the smooth endoplasmic reticulum of the pineal photoreceptors. The intercellular passage of tannic acid mixed with the fixative was blocked at the luminal junctional complex separating the pineal lumen from the basal portion of the pineal epithelium. The passive intercellular transport of substances with high molecular weight from the bloodstream to the cerebrospinal-fluid compartment is thus prevented. However, no blood-brain barrier exists for exogenously administered proteins, which are rapidly taken up by pineal cells and actively transported in a transcellular manner.The findings on the blood-brain barrier of the pineal organ of the rainbow trout are discussed with particular reference to the endocrine capacity of pineal sensory organs.Fellow of the Alexander von Humboldt Foundation, Federal Republic of Germany.     

The pineal organ of Raja clavata: Opsin immunoreactivity and ultrastructure

Summary The pineal organ of Raja clavata was studied by light and electron microscopy, including the immunocytochemical antiopsin reaction. The pineal organ of the ray consists of three portions: (i) a large proximal pineal, (ii) a long tube-like connecting stalk, and (iii) a short distal terminal enlargement. This latter end-vesicle lies in the deep connective tissue layers of the braincase. All portions of the pineal are composed of pinealocytes, intrinsic neurons, ependymal/glial cells, and bundles of nerve fibers embedded in thin neuropil formations. The inner segments of the pinealocytes protrude into the lumen in all parts of the organ and usually contain basal bodies and numerous mitochondria. Often, two outer segments were found to arise from the basal bodies of a single inner segment. By means of light-microscopic immunocytochemistry the outer segments showed a strong antiopsin reaction.The axons of the pinealocytes form ribbon-containing synapses on dendritelike profiles, which appear to belong to the intrinsic pineal neurons. There are other axo-dendritic synapses established by presynaptic terminals lacking ribbons and containing granular and synaptic vesicles. Pineal neurons may contain granular vesicles approximately 60–100 nm in diameter; their processes contribute to the bundles of unmyelinated axons.The fine structural organization of the pineal organ and the opsin immunoreactivity of the outer segments of the pinealocytes indicate a photoreceptive capacity of the organ. The double outer segments represent a peculiar multiplication of the photoreceptor structures.This investigation was supported by grants from the Deutsche Forschungsgemeinschaft to A. Oksche (Ok 1/24; 1/25: Mechanismen biologischer Uhren)On leave from the 2nd Department of Anatomy, Semmelweis OTE, Budapest, Hungary     

Comparative marker analysis of the ependymocytes of the subcommissural organ in four different mammalian species

Summary The subcommissural organ (SCO), classified as one of the circumventricular organs, is composed mainly of modified ependymal cells, attributable to a glial lineage. Nevertheless, in the rat, these cells do not possess glial markers such as glial fibrillary acidic protein (GFAP), protein S100, or the enzyme glutamine synthetase (GS). They receive a synaptic 5-HT input and show pharmacological properties for uptake of GABA resembling the uptake mechanism of neurons. In this study, we examine the phenotype of several mammalian SCO (cat, mouse, rabbit) and compare them with the corresponding features of the rat SCO. In all these species, the SCO ependymocytes possess vimentin as an intermediate filament, but never express GFAP or neurofilament proteins. They do not contain GS as do glial cells involved in GABA metabolism, and when they contain protein S100 (rabbit, mouse), its rate is low in comparison to classical glial or ependymal cells. Thus, these ependymocytes display characteristics that differentiate them from other types of glial cells (astrocytes, epithelial ependymocytes and tanycytes). Striking interspecies differences in the capacity of SCO-ependymocytes for uptake of GABA might be related to their innervation and suggest a species-dependent plasticity in their function.