Activating transcription factor 3 and reactive astrocytes following optic nerve injury in zebrafish

Abstract

Nerve regeneration in the central nervous system is restricted in mammals, but fish and amphibians show amazing resiliency following injury to the central nervous system. We have examined the response of zebrafish (Danio rerio) to optic nerve injury to try to understand the differences between fish and mammals that enable fish to regenerate their optic nerves following crushing and severing. In previous work, we have shown that activating transcription factor 3 (atf3) is expressed at higher levels following optic nerve injury. Here we use a polyclonal anti-ATF3 antibody, anti-cytokeratin (KRT-18) and anti-bystin (BYSL) antibodies to show that Atf3 and Bysl colocalize with cytokeratin-expressing astrocytes in the optic nerve following severing. Furthermore, anti-ATF3 antibodies fail to colocalize with GFP in transgenic zebrafish expressing EGFP in astrocytes Tg(gfap:GFP) or oligodendrocytes Tg(olig2:EGFP). Interestingly, labeling of Atf3 was detected in retinal ganglion cell axons in both the nerve fiber layer and the optic nerve on the injured side. Finally, optic nerve astrocytes labeled with anti-bystin antibodies showed evidence of hypertrophy, suggesting that fish astrocytes in the optic nerve raise a bona fide reactive response to injury even though they do not express glial fibrillary acidic protein.