title: Direct block of calcium channels by dioctanoylglycerol in pregnant rat myometrial cells.
abstract: The effect of 1,2-dioctanoyl-sn-glycerol (DiC8), a diacylglycerol and potent protein kinase C (PKC) activator, on the voltage-dependent slow (L-type) Ca2+ current [lCa(L)] was examined using whole-cell voltage clamp of myometrial cells freshly isolated from late pregnant rats. Bath application of DiC8 (25 microM) decreased ICa(L) by 50.7 +/- 2.4% (n = 22). The effect was reversible and dose dependent (IC50 of 15.3 microM). The effect of DiC8 was not reversed or prevented by application of calphostin C, a selective PKC inhibitor. In addition, 1-oleoyl-2-acetyl-sn-glycerol, another PKC activator, did not produce an inhibitory effect on ICa(L), even at a concentration of 100 microM; ICa(L) was actually slightly stimulated (10.6 +/- 5.1%, n = 6). The steady state inactivation curve for ICa(L) was shifted to the left by DiC8 application (by approximately 15 mV at 25 microM), whereas the activation curve was not affected; the shift should produce a voltage-dependent block. DiC8 decreased ICa(L) progressively during repetitive step depolarizations (use-dependent block). We conclude from these results that (a) DiC8 inhibits ICa(L) independently of PKC in uterine muscle cells and (b) DiC8 preferentially acts on the inactivated state and/or open state of the L-type Ca2+ channels. Therefore, caution must be exercised when studying PKC actions on ion channel regulation using DiC8 as a PKC activator.
authors.affiliation: Department of Molecular and Cellular Physiology, College of Medicine, University of Cincinnati, Cincinnati, Ohio 45267, USA.
authors.forename: M
authors.initials: M
authors.lastname: Kusaka
authors.forename: N
authors.initials: N
authors.lastname: Sperelakis
mesh_terms: D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D004075:Diglycerides / Q000494:pharmacology*; D005260:Female; D009215:Myometrium / Q000187:drug effects* / Q000378:metabolism; D011247:Pregnancy; D011493:Protein Kinase C / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002121:Calcium Channel Blockers; D004075:Diglycerides; C044295:1,2-dioctanoylglycerol; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C