PURPOSE: To examine the expression of c-fos proto-oncogene and phosphorylation of cAMP responsive element binding (CREB) protein in the rat retina after changes in the light-dark condition. METHODS: Rats were exposed to both steady light and flashing light and were killed at the end of light exposure. The retinas were analyzed by in situ hybridization using single-stranded RNA probes for c-fos transcripts and by immunocytochemistry using phosphoSer-133 specific CREB antiserum, anti-calcium calmodulin dependent protein (CaM) kinase II, and anti-CaM kinase IV. RESULTS: c-fos mRNA was expressed in the outer half of the inner nuclear layer (INL) and in the ganglion cell layer (GCL) after 30 minutes of sustained light. After 30 minutes of flashing light, c-fos expression also was detected in the inner border of the INL. Phosphorylated CREB immunoreactive nuclei had similar distribution after steady and flashing light. Both CaM kinase II and CaM kinase IV, which phosphorylate CREB at Ser 133 in vitro, were expressed in the GCL and in the INL. CaM kinase II, however, was localized in the inner border of the INL, whereas CaM kinase IV was distributed in the outer half of the INL. CONCLUSIONS: These results suggest that the differential expression of c-fos mRNA induced by flashing and sustained light may reflect the CREB phosphorylation by CaM kinases in a different subpopulation of retinal cells.
PURPOSE: To examine the expression of c-fos proto-oncogene and phosphorylation of cAMP responsive element binding (CREB) protein in the rat retina after changes in the light-dark condition. METHODS:Rats were exposed to both steady light and flashing light and were killed at the end of light exposure. The retinas were analyzed by in situ hybridization using single-stranded RNA probes for c-fos transcripts and by immunocytochemistry using phosphoSer-133 specific CREB antiserum, anti-calcium calmodulin dependent protein (CaM) kinase II, and anti-CaM kinase IV. RESULTS:c-fos mRNA was expressed in the outer half of the inner nuclear layer (INL) and in the ganglion cell layer (GCL) after 30 minutes of sustained light. After 30 minutes of flashing light, c-fos expression also was detected in the inner border of the INL. Phosphorylated CREB immunoreactive nuclei had similar distribution after steady and flashing light. Both CaM kinase II and CaM kinase IV, which phosphorylate CREB at Ser 133 in vitro, were expressed in the GCL and in the INL. CaM kinase II, however, was localized in the inner border of the INL, whereas CaM kinase IV was distributed in the outer half of the INL. CONCLUSIONS: These results suggest that the differential expression of c-fos mRNA induced by flashing and sustained light may reflect the CREB phosphorylation by CaM kinases in a different subpopulation of retinal cells.