title: Activation of cardiac ryanodine receptors by the calcium channel agonist FPL-64176.
abstract: We investigated the possibility that the Ca(2+) channel agonist FPL-64176 (FPL) might also activate the cardiac sarcoplasmic reticulum (SR) Ca(2+) release channel ryanodine receptor (RyR). The effects of FPL were tested on single channel activity of purified and crude vesicular RyR (RyR2) isolated from human and dog hearts using the planar lipid bilayer technique. FPL (100-200 microM) increased single channel open probability (P(o)) when added to the cytoplasmic side of the channel (P(o) = 0.070 +/- 0.021 in control RyR2; 0.378 +/- 0.086 in 150 microM FPL, n = 9, P < 0.01) by prolonging open times and decreasing closed times without changing current magnitude. FPL had no effect on P(o) when added to the trans (luminal) side of the bilayer (P(o) = 0.079 +/- 0.036 in control and 0.103 +/- 0.066 in FPL, n = 4, no significant difference). The bell-shaped [Ca(2+)] dependence of [(3)H]ryanodine binding and of P(o) was altered by FPL, suggesting that the mechanism by which FPL increases channel activity is by an increase in Ca(2+)-induced activation at low [Ca(2+)] (without a change in threshold) and suppression of Ca(2+)-induced inactivation at high [Ca(2+)]. However, the fact that inactivation was restored at elevated [Ca(2+)] suggests a competitive interaction between Ca(2+) and FPL on inactivation. FPL had no effect on RyR skeletal channels (RyR1), where P(o) was 0.039 +/- 0.005 in control versus 0.030 +/- 0.006 in 150 microM FPL (no significant difference). These results suggest that, in addition to its ability to activate the L-type Ca(2+) channels, FPL activates cardiac RyR2 primarily by reducing the Ca(2+) sensitivity of inactivation.
authors.affiliation: Division of Cardiology, Department of Medicine, and the Feinberg Cardiovascular Research Institute, Northwestern University Medical School, Chicago, Illinois 60611, USA. ja-wasserstrom@northwestern.edu
authors.forename: J Andrew
authors.initials: JA
authors.lastname: Wasserstrom
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Wasserstrom
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Lokuta
authors.forename: James E
authors.initials: JE
authors.lastname: Kelly
authors.forename: Sireen T
authors.initials: ST
authors.lastname: Reddy
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Frank
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002120:Calcium Channel Agonists / Q000494:pharmacology*; D002415:Cats; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D006352:Heart Ventricles / Q000737:chemistry / Q000187:drug effects; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D008861:Microsomes / Q000737:chemistry / Q000378:metabolism; D018482:Muscle, Skeletal / Q000737:chemistry / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism*; D011758:Pyrroles / Q000494:pharmacology*; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects / Q000378:metabolism*
chemical_list: D002120:Calcium Channel Agonists; D011758:Pyrroles; D019837:Ryanodine Receptor Calcium Release Channel; C071658:FPL 64176; D002118:Calcium