Gene profiling of Müller glia during early stages of zebrafish photoreceptor regeneration
ABSTRACT: Photoreceptor damage in adult mammals results in permanent cell loss and glial scarring in the retina. In contrast, adult zebrafish can regenerate photoreceptors following injury. By using a stable transgenic line in which GFP is driven by the cis-regulatory sequences of a glial specific marker gfap, Tg(gfap:GFP)mi2002, previous studies showed that Müller glia, the radial glial cells in the retina, proliferate after photoreceptor loss and give rise to neuronal progenitors that eventually differentiate into regenerated photoreceptors. To identify the molecular mechanisms that initiate this regenerative response, Müller glia were isolated from Tg(gfap:GFP)mi2002 fish during the early stages of regeneration after light lesion and gene expression profiles were generated by microarray analyses. Keywords: time course Retinas were dissected from Tg(gfap:GFP)mi2002 zebrafish at 8, 16, 24, 36 hour post-lesion (hpl) and non-light-treated controls (0 hpl) and were dissociated by enzymatic digestion. GFP+ Müller glia were isolated by fluorescence-activated cell sorting (FACS) for RNA extraction and hybridization on Affymetrix microarrays. Independent hybridization of three biological replicates were performed for each time point.