Regeneration of the newt retina: order of appearance of photoreceptors and ganglion cells.

The adult newt regenerates a functional retina following removal or destruction of the original retina. We studied the order of appearance of cell types in the regenerating retina by using immunohistochemical techniques. An antibody that recognizes the alpha subunit (260 kDa) of voltage-dependent Na+ channels was found to label a 255-kDa band in Western blots of crude membrane fractions from the normal retina. Cryosections of normal retina revealed intense Na+ channel immunoreactivity in somata and axons of ganglion cells, weaker immunoreactivity in somata of amacrine cells, and no immunoreactivity in the inner plexiform layer. In the same sections, immunoreactivity to a monoclonal antibody (RB-1) specific to newt cones was intense in the photoreceptor layer. In regenerating retinas, double staining with the Na+ channel antibody as a possible marker of ganglion cells and RB-1 antibody first revealed immunoreactive cells at the intermediate stage (three to five cells thick), which does not exhibit segregated synaptic layers. Na+ channel-immunoreactive ganglion cells appeared before the RB-1-immunoreactive photoreceptors. Because ganglion cells also appear before photoreceptor cells in normal development, common mechanisms may control both the generation and the regeneration of the newt retina.