Protein kinase C activators inhibit the visual cascade in Limulus ventral photoreceptors at an early stage.
The phosphoinositide cascade mediates visual transduction in invertebrate photoreceptors. Phospholipase C (PLC) catalyzes the hydrolysis of phosphatidylinositol bisphosphate, producing inositol trisphosphate (InsP(3)) and diacylglycerol (DAG). Protein kinase C (PKC) is a major target of DAG in many cell types. We have used PKC activators to investigate the function of the kinase in the phototransduction cascade in Limulus polyphemus ventral photoreceptors. Extracellular application of (-)-indolactam V (0. 03-30 microM) or phorbol-12,13-dibutyrate (10 microM) reversibly reduced the sensitivity of the electrical response of the photoreceptors to light by up to 1000-fold. The inert stereoisomer (+)-indolactam V and 4alpha-phorbol had no effect. The effect of (-)-indolactam V was antagonized by the PKC inhibitors bisindolylmaleimide I and Gö 6976. Coapplication of bisindolylmaleimide V, used as a negative control compound for PKC inhibition, did not reduce the effectiveness of (-)-indolactam V. These findings are consistent with (-)-indolactam V activating PKC and desensitizing the light response. Furthermore, our pharmacological results indicate that PKC activation does not appear to play a role in light adaptation. We localized the position of the target of PKC in the visual cascade. We chemically excited the cascade at various stages to determine the kinase's target. PKC activation by (-)-indolactam V decreased the light-induced elevation of intracellular calcium but had no effect on the photoreceptor's excitatory response to intracellular injection of InsP(3). However, the PKC activator greatly reduced the excitation caused by GTP-gamma-S injection. We propose that PKC inhibits the visual transduction cascade at the G-protein and/or PLC stage.
Dabdoub
A
A
Department of Biology, University of Maryland, College Park, Maryland 20742, USA.
Payne
R
R
D000221:Adaptation, Ocular / Q000187:drug effects; D000818:Animals; D004594:Electrophysiology; D020536:Enzyme Activators / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000502:physiology; D006737:Horseshoe Crabs / Q000502:physiology*; D007211:Indoles / Q000494:pharmacology; D007769:Lactams / Q000494:pharmacology; D009433:Neural Inhibition; D017956:Photoreceptor Cells, Invertebrate / Q000187:drug effects* / Q000502:physiology / Q000528:radiation effects; D011493:Protein Kinase C / Q000378:metabolism*; D010738:Type C Phospholipases / Q000502:physiology; D014785:Vision, Ocular / Q000187:drug effects; D014795:Visual Pathways / Q000187:drug effects* / Q000502:physiology
D020536:Enzyme Activators; D007211:Indoles; D007769:Lactams; C043969:indolactam V; D011493:Protein Kinase C; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins