Inhibition of the calcineurin-like protein phosphatase activity in Limulus ventral eye photoreceptor cells alters the characteristics of the spontaneous quantal bumps and the light-mediated inward currents, and enhances arrestin phosphorylation.
Changes in intracellular calcium are involved in phototransduction processes in both vertebrate and invertebrate photoreceptors. During this phototransduction process in the Limulus ventral eye, there is a biochemical change in the protein phosphatase, calcineurin, such that it becomes capable of activation by calcium and calmodulin. Here we show that the calcium/calmodulin-dependent calcineurin-like activity in light-adapted ventral eye was completely inhibited by the CaN autoinhibitory peptide, CaN A457-482 and the Merck analog of the membrane-permeable, immunosuppressant drug, FK 506, L-683, 590, but not an inactive analogue, L-685, 818. Whole-cell, voltage-clamp recordings of spontaneous quantal bump activity present in dark-adapted photoreceptors injected with either CaN A457-482 (500 microM) or superfused with L-683, 590 (20 microM) or L-685, 818 revealed that both CaN A457-482 and L-683, 590, but not L-685, 818, caused rapid decreases in quantal bump amplitude, rise time and fall time, resulting in smaller, sharper bumps. This was correlated with enhanced phosphorylation of arrestin in light-adapted ventral eye photoreceptors exposed to L-683, 590 or less reliably okadaic acid. Both CaN A457-482 and L-683, 590 markedly affected the light-stimulated inward currents recorded from light-adapted ventral photoreceptors, causing a "terracing" of the inward current, and an intensity-dependent delay in the time required to reach peak amplitude. Consequently, inhibition of calcineurin markedly affects two major rhodopsin-dependent electrophysiological processes, and implicates CaN as an integral component in the phototransduction cascade.
Kass
L
L
Department of Biological Sciences, University of Maine, Orono, USA.
Ellis
D Z
DZ
Pelletier
J
J
Tableman
N E
NE
Edwards
S C
SC
D000818:Animals; D019393:Arrestin / Q000378:metabolism*; D019703:Calcineurin / Q000737:chemistry / Q000494:pharmacology; D065095:Calcineurin Inhibitors; D004594:Electrophysiology; D006737:Horseshoe Crabs / Q000378:metabolism*; D008027:Light; D010446:Peptide Fragments / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D010766:Phosphorylation; D017956:Photoreceptor Cells, Invertebrate / Q000166:cytology / Q000378:metabolism / Q000502:physiology* / Q000528:radiation effects; D016559:Tacrolimus / Q000031:analogs & derivatives / Q000494:pharmacology
D019393:Arrestin; D065095:Calcineurin Inhibitors; D010446:Peptide Fragments; C079136:L 683590; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D016559:Tacrolimus