Expression of neuronal intermediate filament proteins ON1 and ON2 during goldfish optic nerve regeneration: effect of tectal ablation

Goldfish retinal explants were used to study optic tectum participation in the regulation of intermediate filament protein synthesis in retinal ganglion cells during optic nerve regeneration. Retinas were explanted at various times after removal of the contralateral optic tectum. The synthesis of the intermediate filament proteins ON1 and ON2 in the cultures was quantitated by labeling with 35S]methionine, followed by two-dimensional gel electrophoresis, autoradiography, and densitometry. Neuritic growth from the explants was quantitated based on fiber length and density. In retinal explants placed in culture after 23 days of optic nerve regeneration, the synthesis of ON1 and ON2 was reduced when the tectum had been ablated. In contrast, synthesis of these proteins in explants placed in culture at an earlier stage of regeneration was not affected by tectal ablation. At all time points tested, neuritic outgrowth from retinal explants was stimulated by tectal ablation. These findings indicate that the synthesis of the ON1 and ON2 intermediate filament proteins during regeneration is not directly regulated by axonal volume. Further, our findings suggest that interaction between growing axons and tectum is important for sustained expression of these proteins during the later stages of optic nerve regeneration.