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Three proteins, with apparent molecular weights of 16, 30, and 42 kilodaltons, made by glial cells of the goldfish optic nerve undergo enhanced synthesis during regeneration of the retinotectal axons. If, as has been postulated, these enhanced glial proteins (or EGPs) assist in axonal regeneration, they may interact with the retinotectal neurons. Two possible modes of interaction, 1) endocytosis/retrograde transport and 2) binding to optic nerve membranes, were examined using gel electrophoresis of radiolabelled glial proteins. The results suggest 1) that two glial proteins, which may be EGP-16 and EGP-42, are retrogradely transported to the retina during axonal regeneration and 2) that a third glial protein, perhaps EGP-30, binds in a specific and saturable manner to the membranous fraction of the optic nerve.