title: Endothelin-1 and photoreleased diacylglycerol increase L-type Ca2+ current by activation of protein kinase C in rat ventricular myocytes.
abstract: The amphotericin B-perforated whole-cell patch clamp technique was used to determine the modulation of L-type Ca2+ channels by protein kinase C (PKC)-mediated pathways in adult rat ventricular myocytes. Application of 10 nM endothelin-1 (ET-1) increased peak Ca2+ current (ICa) by 28.2 +/- 2.5 % (n = 13) and slowed current decay. These effects were prevented by the endothelin receptor antagonist PD145065 (10 microM) and by the PKC inhibitor chelerythrine (8 microM). To establish if direct activation of PKC mimicked the ET-1 effect, the active and inactive phorbol esters (phorbol-12-myristate-13-acetate and 4alpha-phorbol-12, 13-didecanoate) were tested. Both phorbol esters (100 nM) resulted in a small (approximately 10%) increase in ICa, suggesting PKC-independent effects. Bath application of dioctanoylglycerol (diC8), a diacylglycerol (DAG) analogue which is capable of directly activating PKC, caused a gradual decline in peak ICa (50.4 +/- 6.2 %, n = 5) and increased the rate of current decay. These effects were unaffected by the PKC inhibitor chelerythrine (8 microM). Intracellular photorelease of caged diC8 with 3 or 10 s exposure to UV light produced a concentration-dependent increase in peak ICa (20. 7 +/- 8.5 % (n = 8) for 3 s UV and 60.8 +/- 11.4 % (n = 13) for 10 s UV), which could be inhibited by chelerythrine. Our results demonstrate that both ET-1 and intracellularly photoreleased diC8 increase ICa by a PKC-mediated pathway, which is in direct contrast to the PKC-independent inhibition of ICa produced by bath-applied diC8. We conclude that specific cellular pools of DAG are crucially important in the regulation of ICa by PKC.
authors.affiliation: Department of Medicine, University of Wisconsin-Madison, Madison, WI 53792, USA.
authors.forename: J Q
authors.initials: JQ
authors.lastname: He
authors.forename: Y
authors.initials: Y
authors.lastname: Pi
authors.forename: J W
authors.initials: JW
authors.lastname: Walker
authors.forename: T J
authors.initials: TJ
authors.lastname: Kamp
mesh_terms: D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D002104:Cadmium / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D002273:Carcinogens / Q000494:pharmacology; D004075:Diglycerides / Q000493:pharmacokinetics*; D004594:Electrophysiology; D019332:Endothelin-1 / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000201:enzymology; D009206:Myocardium / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology; D009543:Nifedipine / Q000494:pharmacology; D010617:Phenanthridines / Q000494:pharmacology; D010777:Photochemistry; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014466:Ultraviolet Rays
chemical_list: D000470:Alkaloids; D053119:Benzophenanthridines; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D002273:Carcinogens; D004075:Diglycerides; D019332:Endothelin-1; D004791:Enzyme Inhibitors; D010617:Phenanthridines; D002104:Cadmium; C044295:1,2-dioctanoylglycerol; C016299:chelerythrine; D011493:Protein Kinase C; D009543:Nifedipine; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium