Activity-dependent sharpening of the regenerating retinotectal projection in goldfish: relationship to the expression of growth-associated proteins

Abstract

During regeneration of the optic nerve in goldfish, manipulations that disrupt the transmission of patterned visual information, if applied within the so-called ‘sensitive period’, lead to the formation of a diffuse retinotopic map (Schmidt, Cell. Mol. Neurobiol., 5 (1985) 65). The present study examined: (a) whether the sensitive period (14–50 days postcrush) coincides with the period in which specific ‘growth-associated proteins’ are present in the regenerating optic nerve terminals; and (b) whether manipulations that alter physiological activity during the sensitive period influence the expression of these proteins. Following bilateral optic nerve crush, goldfish regenerated their optic nerves either under normal illumination conditions (control), in total darkness, or with physiological activity suppressed in the nerve by intraocular injections of tetrodotoxin (TTX). At various times postcrush, proteins conveyed from the retina to the developing nerve endings were visualized by labeling the eye with [ 35S]methionine and then analyzing, by 2-dimensional gel electrophoresis and fluorography, radiolabeled proteins present in the optic tectum 15 h later. Rapidly-transported proteins that underwent large, specific increases during regeneration included the previously described 48 kDa growth-associated protein (GAP-48); labeling of GAP-48 was maximal during axonal outgrowth and then declined, but still remained well above background levels throughout the ‘sensitive period’. Another group of rapidly-transported proteins, mol. wt. = 110–140 kDa (HMW), followed a similar time course, while levels of a 28 kDa protein peaked at 2 weeks and then declined rapidly. Thus, activity-dependent ‘sharpening’ processes occur during a period in which the levels of GAP-48 and HMW remain elevated in the nerve terminals. Dark-rearing and TTX were found to alter somewhat the synthesis and axonal transport of these proteins at early, but not later portions of the sensitive period.