Antigenic properties of large ganglion cells isolated from ox retina

Abstract

Antisera were prepared against large ganglion cells (>28 μm diameter) that were isolated from ox retina. The cells of the retina were dispersed with a cytosieve and a nylon cloth and then isolated by centrifugation in a discontinuous sucrose density gradient. Rabbits were injected subcutaneously with the isolated cells suspended in complete Freund's adjuvant and then injected intravenously with the cells suspended in a sucrose solution. Antibodies were obtained which formed precipitin lines in Ouchterlony plates when diffused against Triton X-100 extracts of large ganglion cells and which agglutinated selectively the large ganglion cells from retinal dispersions. The specificity of the antisera for the retinal cell types was examined using immunofluorescence and immunoperoxidase methods. Immunoglobulins reacted most intensely with the large ganglion cells and their projections in the internal plexiform layer of retinal sections. An intense reaction within the external plexiform layer was also observed. Although the antisera were prepared against the large ganglion cells from ox retinas, 6 μm thick sections of ox, dog, cat, rabbit, macaque, rat and goldfish retinas reacted similarly with the immunoglobulins, indicating that the antigens were not species-specific. A much weaker reaction was observed between the antisera and the smaller ganglion cells, the amacrine cell bodies and the cone receptor cells. The immunuglobulins also reacted with the granule and Purkinje cells in the cerebellar cortex. One predominant retinal antigen was isolated by affinity chromatography; this protein or subunit had a molecular weight of 49000 daltons as determined by electrophoresis on polyacrylamide gels containing sodium dodecylsulfate.