Reissner's Fiber Research Articles

Neuraminidase injected into the cerebrospinal fluid impairs the assembly of the glycoproteins secreted by the subcommissural organ preventing the formation of Reissner's fiber.

Neuraminidase was injected into the cerebrospinal fluid of normal rats to investigate the assembly and fate of the desialylated Reissner's fiber glycoproteins. It was established that a single injection of neuraminidase cleaved the sialic acid residues of the Reissner's fiber glycoproteins that had been assembled before the injection, and of the molecules that were released over a period of at least 4 h after the injection. These desialylated glycoproteins underwent an abnormal assembly that led to the formation of spheres instead of a fiber. The number of these spheres increased during the 4-h period following the injection, indicating that neuraminidase did not prevent the secretion of the Reissner's fiber glycoproteins into the cerebrospinal fluid. The spheres remained attached to the surface of the subcommissural organ and became intermingled with infiltrating cells, many of which were immunocytochemically identified as macrophages. The latter were seen to contain immunoreactive Reissner's fiber material. It is concluded that the desialylated Reissner's fiber glycoproteins forming the spheres underwent an in situ degradation by macrophages, thus resembling the normal process undergone by the Reissner's fiber glycoproteins reaching the massa caudalis.

Complex expression pattern of the SCO-spondin gene in the bovine subcommissural organ: toward an explanation for Reissner's fiber complexity?

Bovine SCO-spondin is a glycoprotein secreted by the subcommissural organ (SCO), an ependymal derivative located in the roof of the third ventricle. It shows homology with developmental molecules involved in directional axonal growth. Using SCO-spondin cDNAs as probes, we analysed the specific expression of the corresponding gene in the bovine SCO by Northern blot and in situ hybridization (ISH). A strong expression was detected in the secretory ependymal and hypendymal cells of the SCO and the main transcripts showed a large size 14 kb. A single copy gene was revealed by Southern blot analysis of bovine genomic DNA. The presence of additional transcripts suggested a transcriptional regulation of the SCO-spondin gene. A comparative analysis of the results obtained by molecular and immunological techniques (immunoblotting and immunopurification) pointed to the presence of several SCO-spondin related proteins in the SCO encoded by the same gene. The presence in the cerebral hemispheres (CH) of a 54-kDa glycoprotein with a common epitope is discussed as a putative cleaved SCO-spondin product carried by the cerebrospinal fluid, that may act on neuronal development.

Spontaneous congenital hydrocephalus in the mutant mouse hyh. Changes in the ventricular system and the subcommissural organ.

The subcommissural organ is an ependymal gland located at the entrance of the cerebral aqueduct. It secretes glycoproteins into the cerebrospinal fluid, where they aggregate to form Reissner's fiber. This fiber grows along the aqueduct, fourth ventricle, and central canal. There is evidence that the subcommissural organ is involved in the pathogenesis of congenital hydrocephalus. This organ was investigated in the mutant mouse hyh developing a congenital hydrocephalus. The central nervous system of normal and hydrocephalic hyh mice, 1 to 40 days old, was investigated using antibodies recognizing the subcommissural organ secretory glycoproteins, and by transmission and scanning electron microscopy. At birth, the affected mice displayed open communications between all ventricles, absence of a central canal in the spinal cord, ependymal denudation of the ventricles, stenosis of the rostral end of the aqueduct, and hydrocephalus of the lateral and third ventricles and of the caudal end of the aqueduct. Around the 5th postnatal day, the communication between the caudal aqueduct and fourth ventricle sealed, and hydrocephalus became severe. It is postulated that the hyh mice carry a genetic defect affecting the ependymal cell lineage. The subcommissural organ showed signs of increased secretory activity; it released to the stenosed aqueduct a material that aggregated, but it did not form a Reissner's fiber. A large area of the third ventricular wall differentiated into a secretory ependyma synthesizing a material similar to that secreted by the subcommissural organ. It is concluded that the subcommissural organ changes during hydrocephalus; whether these changes precede hydrocephalus needs to be investigated.

Mindin/F-spondin Family: Novel ECM Proteins Expressed in the Zebrafish Embryonic Axis

F-spondin is a secreted protein expressed at high levels by the floor plate cells. The C-terminal half of the protein contains six thrombospondin type 1 repeats, while the N-terminal half exhibited virtually no similarity to any other protein until recently, when aDrosophilagene termedM-spondinwas cloned; its product was found to share two conserved domains with the N-terminal half of F-spondin. We report the molecular cloning of four zebrafish genes encoding secreted proteins with these conserved domains. Two are zebrafish homologs ofF-spondin,while the other two, termedmindin1andmindin2,encode mutually related novel proteins, which are more related to theDrosophilaM-spondin than to F-spondin. During embryonic development, all four genes are expressed in the floor plate cells. In addition to the floor plate,mindin1is expressed in the hypochord cells, whilemindin2is expressed in the sclerotome cells. When ectopically expressed, Mindin proteins selectively accumulate in the basal lamina, suggesting that Mindins are extracellular matrix (ECM) proteins with high affinity to the basal lamina. We also report the spatial distribution of one of the F-spondin proteins, F-spondin2. F-spondin2 is localized to the thread-like structure in the central canal of the spinal cord, which is likely to correspond to Reissner's fiber known to be present in the vertebrate phylum. In summary, our study has defined a novel gene family of ECM molecules in the vertebrate, all of which may potentially be involved in development of the midline structure.

In vitro differentiation of chick spinal cord neurons in the presence of Reissner's fibre, an ependymal brain secretion

The subcommissural organ (SCO), which belongs to the circumventricular organs, is a specialized ependymal structure of the brain that secretes glycoproteins into the cerebrospinal fluid (CSF) which condense to form a thread-like structure, the Reissner's fibre (RF). Regarding the presence of this ependymal brain secretion all along the central canal of the developing spinal cord, we analysed a putative developmental activity of RF on neuronal spinal cord cells. The effects of RF proper and soluble RF-material were examined in primary cultures of dissociated spinal cord cells from day 6 chicken embryos. In serum-containing mixed glial/neuronal cell cultures, both RF and soluble RF-material promoted neuronal survival. This effect was blocked by addition of specific antibodies raised against bovine RF into the culture medium. In serum-free neuron-enriched cultures, no neuronal survival activity was observed; however, under these conditions RF proper induced neuronal aggregation and neuritic outgrowth of spinal cord cells. Interestingly, neurites extending from the aggregates appeared mainly unfasciculated. Our results suggest a direct modulation of cell–cell interactions by SCO/RF glycoproteins and an indirect survival effect on neurons. These data strengthen the hypothesis of the involvement of SCO/RF complex in the development of the central nervous system (CNS) and are discussed regarding molecular features of SCO-spondin, a novel glycoprotein recently identified in this complex.

Cervical cord ependymoma with numerous microrosettes

"Microrosette ependymoma," which is ependymoma with numerous microrosettes throughout the tumor, has rarely been reported. We describe an autopsy case of cervical cord ependymoma with two unusual features: the presence of numerous microrosettes and the formation of trabecular architecture. The tumor originated in the C2 segment of a man aged 23 years and gradually expanded over the following 15 years and 10 months until the entire cervical cord was involved. Beside the low grade of malignancy, the tumor cells exhibited a strong tendency to form microrosettes and trabecular architecture, which formed many perivascular pseudorosettes. The microrosettes mostly consisted of only two or a few more cells, in the absence of large rosettes. Thus the constituent cells were those forming perivascular pseudorosettes. Electron microscopy and immunohistochemistry characterized the ependymal properties of the microrosettes, whose lumina frequently contained fibril bundles similar to those of the Reissner's fiber fibrils, in addition to cilia and microvilli. The pathogenesis of the occurrence of numerous microrosettes is unknown; however, a defect in the mechanism of regulation of rosette formation and enlargement is the most likely explanation.

Production of an oxytocin like substance by the subcommissural organ (SCO), related to the reproductive cycle in oviparous and viviparous reptiles.

The subcommissural organ (SCO) is a circumventricular organ of glial origin typical of all vertebrates. The SCO releases its secretion into the third ventricle to constitute Reissner's fibre (RF). Reportedly, in reptiles, SCO has cyclic secretory activity related to the reproductive cycle. In this immunocytochemical study we show that, in females of oviparous reptiles (Lacertidae: Podarcis sicula) and in a viviparous species (Scincidae: Chalcides chalcides), SCO secretion consists of hormones, in part of the oxytocin-like (OXY-like) type. The amount of OXY-like material in the cells and in the third ventricle varies according to the different stages of the reproductive cycle. In the oviparous species, OXY-like hormone secretion can be induced by FSH administration at 28 degrees C, in the period of winter reproductive stasis as well. In the viviparous skink, showing an annual single ovulatory cycle, OXY-like secretion is present in the basal region of the cells, and is released into the third ventricle only at delivery. The role of an OXY-like hormone in the SCO is here discussed in relation to the different stages of the reproductive cycle. Its influence on the hypothalamus-hypophysis-gonad axis and its role in the transport of eggs into the ducts in the oviparous species, and at delivery in the viviparous one, are also suggested.

Rostral floor plate (flexural organ) secretes glycoproteins immunologically similar to subcommissural organ glycoproteins in dogfish ( Scyliorhinus canicula) embryos

The subcommissural organ of vertebrates secretes glycoproteins into the cerebrospinal fluid of the third cerebral ventricle. This material polymerizes in Reissner's fiber. During ontogenetic development, besides the subcommissural organ, the ependyma lining the pontine flexure constitutes an additional Reissner's fiber-secreting gland named flexural organ. We have studied the secretion of the flexural organ and the subcommissural organ in dogfish ( Scyliorhinus canicula) embryos using three different antisera and the lectins concanavalin A and wheat germ agglutinin. AFRU is an antiserum against the bovine Reissner's fiber; Ab-600 is an antiserum against 600 kDa dogfish subcommissural organ glycoproteins; and APSO is an antiserum against immunoaffinity purified bovine subcommissural organ secretory glycoproteins. These three antisera immunostained the flexural organ indicating that it contains epitopes similar to those present in bovine and dogfish subcommissural organ glycoproteins. It seems highly probable that the flexural organ and the subcommissural organ of dogfish embryos secrete similar compound(s). Other ependymal regions were also immunostained with Ab-600 and APSO antisera. Then, Reissner's fiber-like glycoproteins were transiently expressed by most embryonary ependymal cells. These glycoproteins might play a role in the development of the central nervous system of vertebrates.

Calcitonin-like immunoreactivity in the subcommissural organ and Reissner's fiber in the teleost Clarias batrachus, frog Rana tigrina and lizard Calotes versicolor

In the CNS of vertebrates, although the subcommissural organ (SCO) has been identified as an ependymal brain gland and Reissner's fiber (RF) as a condensed product of its secretion, the exact nature of the secretory substances has remained elusive. In the present study, immunocytochemical application of polyclonal antibodies against calcitonin revealed intense immunoreactivity in the cells, cell processes and cerebrospinal spinal fluid (CSF)-contacting apical terminals of the columnar ependymal cells of the SCO in the teleost, Clarias batrachus. Intense immunoreactivity was also seen throughout the length of the RF as it extended along the Sylvian aqueduct, fourth ventricle and central canal of the spinal cord. Control procedures were employed to confirm the specificity of the immunoreaction. The results for the first time suggest that calcitonin-like substance may be the synthetic and secretory product of the SCO that may be released into the CSF or stored in the RF. Presence of calcitonin-like immunoreactivity in the SCO-RF complex of the frog Rana tigrina and the lizard Calotes versicolor underscores wider significance of the phenomenon. In view of the potentials of these findings, it is felt that investigations aimed at establishing the precise nature of calcitonin-like immunoreative material in the SCO-RF complex may be rewarding. © 1997 Elsevier Science B.V. All rights reserved.

Reissner's fibre promotes neuronal aggregation and influences neuritic outgrowth in vitro.

Reissner's fibre is the condensed form of glycoproteins secreted by the subcommissural organ; it extends through the central canal to the caudal end of the spinal cord. The effect of Reissner's fibre was assessed on dissociated embryonic chick cortical neuronal cells grown in chemically defined medium, by using two cell culture systems: (1) low-density cultures, in which neuronal cells remained evenly distributed; (2) high-density cultures, in which neuronal cells aggregated, displaying prominent neuritic outgrowth. Reissner's fibre, when added to low-density cultures, induced neuronal aggregation and neuritic outgrowth but this effect was restricted to an area centred around Reissner's fibre. Reissner's fibre, when added to high-density cultures, potentiated cell aggregation. Antibodies inhibiting the neural cell adhesion molecule or N-cadherin, and soluble Reissner's fibre material (reported previously to have anti-aggregative activity), did not prevent the aggregative activity induced by Reissner's fibre. Neuronal cells showed a similar reaction pattern when heparin or Reissner's fibre was added to the culture. These results suggest that the subcommissural organ/Reissner's fibre complex has multifunctional activities and may modulate cell-cell interactions during the development of the central nervous system.

Identification of a high molecular weight polypeptide in the subcommissural organ of the chick embryo.

The subcommissural organ is an ependymal brain gland that secretes, into the ventricular cerebrospinal fluid, high molecular weight glycoproteins that form Reissner's fiber. Precursor and processed forms of secretion have been demonstrated by immunoblotting in the subcommissural organ of mammals and fish. In the chicken only a processed form has as yet been identified. In the present report, we have studied the subcommissural organ of 13-day-old chick embryos using (1) an antiserum against bovine Reissner's fiber, and (2) the lectins, concanavalin A and Limax flavus agglutinin. Paraffin sections of the subcommissural organ and blots of subcommissural organ extracts have been analyzed. The ependymal cells of sectioned subcommissural organ are strongly stained with the antiserum. Concanavalin A binds to materials in all cytoplasmatic regions, whereas Limax flavus agglutinin identifies materials confined to the apex of the ependymal cells. In the blots, a band of 540 kDa is immunostained. This band is positive for concanavalin A positive but negative for Limax flavus agglutinin and is thereby regarded as representing a precursor form of the secretion.

Soluble material from Reissner's fiber displays anti-aggregative activity in primary cultures of chick cortical neurons

The subcommissural organ (SCO), which belongs to the circumventricular organs, is a specialized ependymal structure of the brain that secretes glycoproteins into the cerebrospinal fluid (CSF) which condense to form a thread-like structure, Reissner's fiber (RF). The effects of soluble material released by RF were examined in primary cultures of dissociated cortical cells from embryonic (day 8) chick brain. Under serum-free conditions, the presence in the cultures of soluble RF material markedly impaired neuronal cell aggregation. This effect was completely blocked by addition into the culture medium of specific antibodies raised against bovine RF. The anti-aggregative effect of soluble RF material is observed on poly- l -lysine as well as on diffrrent extracellular matrix proteins including collagen and laminin, but was less effective on fibronectin. The continuous exposure of the cultures to soluble RF material for 7 days significantly decreased choline acetyltransferase activity. On the other hand, soluble RF material did not appear to have mitogenic activity on neuronal cultures. Modulation of cell-cell interactions by SCO/RF glycoproteins sterngthens the hypothesis of the involvement of RF in developmental events of the central nervous system.

Soluble material from Reissner's fiber displays anti-aggregative activity in primary cultures of chick cortical neurons

The subcommissural organ (SCO), which belongs to the circumventricular organs, is a specialized ependymal structure of the brain that secretes glycoproteins into the cerebrospinal fluid (CSF) which condense to form a thread-like structure, Reissner's fiber (RF). The effects of soluble material released by RF were examined in primary cultures of dissociated cortical cells from embryonic (day 8) chick brain. Under serum-free conditions, the presence in the cultures of soluble RF material markedly impaired neuronal cell aggregation. This effect was completely blocked by addition into the culture medium of specific antibodies raised against bovine RF. The anti-aggregative effect of soluble RF material is observed on poly- l-lysine as well as on diffrrent extracellular matrix proteins including collagen and laminin, but was less effective on fibronectin. The continuous exposure of the cultures to soluble RF material for 7 days significantly decreased choline acetyltransferase activity. On the other hand, soluble RF material did not appear to have mitogenic activity on neuronal cultures. Modulation of cell-cell interactions by SCO/RF glycoproteins sterngthens the hypothesis of the involvement of RF in developmental events of the central nervous system.

Radial secretory glia conserved in the postnatal vertebrate brain: a study in the rat.

Secretory glial cells in the roof of the last diencephalic prosomer, ependymocytes and hypendymocytes, form the subcommissural organ. The cells of this complex were labelled immunocytochemically, using an antiserum against their specific secretory products. The study aims at the characterization of this cell type in the rat as an anatomical model situation. Radially oriented secretory glial cells remain after birth behind the posterior commissure in the mesencephalic aqueduct. At about postnatal day 10, the cell bodies descend into the conventional ependyma and at postnatal day 25 they assume a compact, rounded appearance. The secretory product they release is involved in the formation of Reissner's fiber. This differentiation in phenotype is not accompanied by a change of the intermediate filament expression. In the adult rat these cells had been labelled immunopositive for cytokeratins 8 and 18 as well as vimentin but not for glial fibrillary acidic protein. DiI-marking from the third ventricle and from the dorsal surface of the brain shows that the basal processes of ependymocytes and hypendymocytes project to the external and internal glial limiting membrane, respectively, through the commissural fiber bundles. Also the subependymal located hypendymocytes have apical processes with contacts to the cerebrospinal fluid. When this secretory cell population is studied with respect to cyto-architectonical changes during ontogeny the results lead to a new understanding of the subcommissural cells. They are not specialized ependymal cells in a regionally restricted and secondary differentiated ependymal area, but rather descendants of an ontogenetically ancient, specific type of radial glia. Characteristic features for all subcommissural cells are that they: (1) appear very early during ontogeny, (2) are derived from a radial oriented glial cell type, (3) carry at least one kinocilium, (4) possess an original intermediate filament pattern, (5) release a secretory product.

Bovine Reissner's fiber (RF) and the central canal of the spinal cord: an immunocytochemical study using a set of monoclonal antibodies against the RF-glycoproteins.

The subcommissural organ secretes N-linked complex-type glycoproteins into the cerebrospinal fluid. These glycoproteins condense to form Reissner's fiber (RF), which extends along the fourth ventricle and central canal of the spinal cord. A set of three monoclonal antibodies (Mabs 3E6, 3B1, and 2A5) has been obtained using these glycoproteins as immunogens. Competitive and sandwich enzyme-linked immunoassay methods have demonstrated that the three monoclonal antibodies are directed against different epitopes, and that there is no competition among them for their binding to glycoproteins of RF. Mab 3E6 displays immunoblotting properties that are similar to those of a polyclonal antibody against the pool of glycoproteins from RF, but that are different from those of Mabs 3B1 and 2A5. All three antibodies immunostain the bovine subcommissural organ and RF. A population of ependymal cells is stained by the polyclonal antibody, and Mabs 2A5 and 3E6, but not by Mab 3B1. The material present in a population of ependymal cells of the central canal, and the glycoproteins secreted by the subcommissural organ thus probably have partial chemical identity. Some evidence suggests that the immunoreactive ependymal cells are secretory cells. The luminal surface of the central canal is coated by a thin layer of material with immunocytochemical characteristics different from those of the ependymal cells; such a coat may correspond to material released from RF.

Immunochemical analysis of the subcommissural organ-Reissner's fiber complex using antibodies against alkylated and deglycosylated glycoproteins of the bovine Reissner's fiber

Reissner's fiber (RF) has been isolated, solubilized, and used to raise polyclonal antibodies. The present investigation has been designed: (1) to obtain antibodies against RF-glycoproteins in their native form (anti-RF-BI), after irreversible denaturation by alkylation (anti-RF-A), and after alkylation and deglycosylation by using endoglycosidase F (anti-RF-DE); (2) to use these antisera for a comparative immunocytochemical study of the subcommissural organ (SCO)-RF complex; (3) to establish the molecular mass of the deglycosylated RF-glycoproteins. Anti-RF-BI reacts with the SCO of all the species investigated. Anti-RF-A and anti-RF-DE only react with bovine SCO and RF. The three antisera stain the same bands in immunoblots of extracts of bovine SCO and RF, but anti-RF-A and anti-RF-DE reveal additional bands. The epitope common to all species reacting with anti-RF-BI is thus probably conformational in nature and associated with the integrity of the disulfide bonds. The lack of antibodies against conserved sequential epitopes in anti-RF-A does not support previous assumptions on the conserved nature of the SCO secretion. After deglycosylation by using endoglycosidase F, the RF-glycoproteins present a reduction in their molecular mass ranging between 10% and 25%. The three larger compounds (450, 300, and 230 kDa) lose their affinity for Limax flavus agglutinin (affinity = sialic acid), whereas the 190-kDa compound (170 kDa after deglycosylation) keeps its affinity for this lectin suggesting that it has N-linked and O-linked carbohydrate moieties, the three larger proteins probably having only N-linked carbohydrates.

SCO-spondin: a new member of the thrombospondin family secreted by the subcommissural organ is a candidate in the modulation of neuronal aggregation

A number of cues are known to influence neuronal development including growth factors, cell-adhesion molecules, components of the extracellular matrix and guidance molecules. In this study, we present molecular and functional evidence that SCO-spondin, a novel relative of the thrombospondin family, could also be involved in neuronal development by modulating cell aggregative mechanisms. SCO-spondin corresponds to glycoproteins secreted by the subcommissural organ (SCO), an ependymal differentiation of the vertebrate brain located at the entrance to the Sylvian aqueduct. A cDNA clone of 2.6 kb, isolated from a bovine SCO cDNA library, was shown to be specifically and highly expressed in the bovine SCO by in situ hybridization and was subsequently sequenced. Analysis of the deduced amino acid sequence reveals the presence of four conserved domains known as thrombospondin (TSP) type I repeats. To account for the homology with thrombospondins and F-spondin, this secreted glycoprotein was called SCO-spondin. Two potent binding sites to glycosaminoglycan (BBXB) and to cytokine (TXWSXWS) are also found in the TSP type I repeats. The deduced amino acid sequence exhibits three other conserved domains called low density lipoprotein (LDL) receptor type A repeats. The possibility of SCO-spondin involvement in neuronal development as a component of the extracellular matrix is discussed regarding these molecular features. The idea of a modulation of cell-cell and/or cell-matrix interaction is further supported by the anti-aggregative effect observed on cultured neuronal cells of material solubilized from Reissner's fiber. That Reissner's fiber, the condensed secretory product of the SCO present along the whole spinal cord can be a potent morphogenetical structure is an important concept for the analysis of the molecular mechanisms leading to spinal cord differentiation.

The Subcommissural Organ and Reissner's Fiber Complex: An Enigma in the Central Nervous System?

The Subcommissural Organ and Reissner's Fiber Complex: An Enigma in the Central Nervous System?

Reissner's fibre supports the survival of chick cortical neurons in primary mixed cultures

Reissner's fibre supports the survival of chick cortical neurons in primary mixed cultures

Light- and electron-microscopic immunocytochemical investigation of the subcommissural organ using a set of monoclonal antibodies against the bovine Reissner's fiber

Ten monoclonal antibodies (Mabs) against glycoproteins of the bovine Reissner's fiber (RF) have been used in a structural and ultrastructural immunocytochemical investigation of the bovine subcommissural organ (SCO) and RF. The SCO of other vertebrate species has also been studied. For comparison, polyclonal antibodies against bovine RF (AFRU) were used. The SCO and RF of ox, pig and dogfish and the SCO of dog, rabbit, rat and frog were submitted to light-microscopic immunocytochemistry using AFRU and Mabs. Postembedding ultrastructural immunocytochemistry was applied to sections of bovine SCO using AFRU and Mabs. Bovine SCO consists of ependymal and hypendymal cell layers, the latter being arranged as cell strands across the posterior commissure, or as hypendymal rosette-like structures. All cytoplasmic regions of the ependymal and hypendymal cells were strongly stained with AFRU. Six Mabs showed the same staining pattern as AFRU, one Mab stained RF strongly and SCO weakly, two Mabs stained RF but not SCO, and, finally, one Mab (3B1) exclusively stained the apices of the ependymal and hypendymal cells. All Mabs recognized the SCO and RF of the pig. Two Mabs bound to the SCO of the dog. One Mab stained the SCO of the rabbit and another the SCO of the rat. The SCO of frog and dogfish were totally negative. Bovine SCO stained with AFRU, showed label in the rough endoplasmic reticulum (RER) and the secretory granules (SG) of the ependymal and hypendymal cells. The former, in the form of parallel cisternae, reticulum or concentric rings, was seen throughout all cytoplasmic regions. SG were abundant in the apical pole of the ependymal and hypendymal cells. Only one Mab showed a staining pattern similar to AFRU. Five Mabs showed strong reactions in the SG but weak labeling of the RER. Mab 3B1 showed the label confined to the SG only. Our results suggest that: (i) in the bovine tissue, some epitopes are present in both precursor and processed materials, whereas others are characteristic of mature glycoproteins present in SG and the RF; (ii) the bovine SCO secretes at least two different compounds present in ependymal and hypendymal cells; (iii) both compounds coexist in the same secretory granule; (iv) there are conserved, class-specific, and species-specific epitopes in the glycoproteins secreted by the SCO of vertebrates.