The protein-protein interactions that underlie shut-off of the light-activated rhodopsin were studied using synthetic peptides derived from C-terminal region of the rhodopsin. The photoresponses were recorded in whole-cell voltage clamp from rod outer segments (ROS) that were internally dialyzed with an intracellular solution containing the synthetic peptides. This was the first time that synthetic peptides have been used in functionally intact ROS. None of the tested peptides promoted the shut-off of the photolyzed rhodopsin (R*) by stimulating the binding of an activated arrestin to non-phosphorylated R*, contrary to what was expected from in vitro experiments (Puig et at. FEBS Lett. 362: 185-188, 1995).
Protein kinase C and polyphosphoinositide metabolism are reported to affect light-activated processes in cell free systems. To investigate their role in phototransduction under more physiological conditions the effects of nonhydrolyzable inositol trisphosphate (IP3) analogs as well as of protein kinase C and phospholipase C inhibitors on the characteristics of the electrical light response were studied. Rod outer segments were dialyzed in whole-cell voltage clamp and photoresponses in the presence and absence of the tested compounds were compared. None of the compounds influenced the light responses suggesting that neither IP3 nor protein kinase C participate in the phototransduction cascade. A number of different proposals about the participation of protein kinase C and inositol trisphosphate (1P3) in the phototransduction process based on a wide variety of in vitro experiments should therefore be reevaluated.
Vision is a fascinating example of the interaction of a biological system with the outside world. The first step of translating electromagnetic energy into a biologically recognizable signal involves the phototransduction cascade in retinal photoreceptor cells. Phototransduction is the best studied example of a GTP binding protein (G protein)- coupled signal transduction pathway. A great body of knowledge about phototransduction has been established in the past several decades but there are still many unanswered questions, particularly about photoresponse recovery and adaptation. The purpose of this review is to outline the events following photon absorption by vertebrate photoreceptors, to demonstrate the great complexity of the phototransduction cascade mechanisms, and to point out some of the controversies arising from recent fmdings in the field of visual transduction.