Antibodies were raised in rabbit against crude subcommissural organ (SCO) extract of 19 day old chick embryos. After absorption with crude brain extract, the IgG fraction was purified by ion exchange chromatography. The specificity of the antibodies was controlled by immunostaining and by a competition test between lectins (Concanavalin A-Con A- and wheat germ agglutinin-WGA-) and antibodies (A74 IgG). Using A74 IgG, some ependymal cells containing immunoreactive material (IRM) could be detected in the SCO anlage at 4 days of incubation. During the following stages (5 to 12 days), the immunostaining extended caudal-ward in the SCO epithelium according to a rostro-caudal gradient of differentiation. The appearance of IRM in the secretory ependymal cells of the SCO parallel that of Concanavalin A-positive glycoproteins (Bruel et al., 1987). Secretion of IRM into the ventricular cavity, contributing to the formation of Reissner's fiber (RF) occurred during the 7th day of incubation. The formation of RF was examined at different levels of the spinal cord using A74 IgG, WGA and aldehyde fuchsin (AF) staining. The appearance of SCO specific glycoproteins was observed at 11 days in the central canal but the presence of a non-immunoreactive material at 10 days suggests that the formation of RF probably happens inside a guidance material.
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http://dx.doi.org/10.1007/BF01245126 | DOI Listing |
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