Abstract
Several isomers of hexachlorocyclohexanes (HCHs) have been shown to be toxic to mammals. Previous studies have revealed that the δ isomer (δ-HCH) was particularly potent toward disrupting Ca2+homeostasis in a variety of excitable and nonexcitable cells and altering contractility of cardiac muscle. The effects of the δ and γ isomers of HCH were further investigated on isolated ventricular myocytes from guinea pig and on single cardiac ryanodine receptor (RyR2) Ca2+-release channels from cardiac SR vesicles. Intracellular Ca2+ transients were examined in electrically stimulated cells using the fluorescent dye indo-1, and twitch contractions of myocytes were analyzed using a video-based edge motion detection system. Exposure of myocytes to δ- but not γ-HCH depressed the peak of intracellular Ca2+ transients and prolonged recovery time. These effects were correlated with the ability of δ-HCH to inhibit the binding of [3H]ryanodine, a conformationally sensitive probe for RyR2 function, to SR preparations (IC50 = 2 and 18 μM for high- and low-affinity interactions, respectively). Measurements of single-channel gating kinetics under voltage-clamp provided direct evidence of a potent isoform-selective activation of RyR2 by δ-HCH. Results from these studies revealed that δ-HCH alters Ca2+ homeostasis and contractility in cardiac myocytes and that the mechanism can be ascribed, at least in part, to a direct interaction with the RyR2 channel complex.
Footnotes
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Send reprint requests to: Isaac N. Pessah, Ph.D., Department of Molecular Biosciences, School of Veterinary Medicine, University of California, 1 Shields Ave., Davis, CA 95616. E-mail:inpessah{at}ucdavis.edu
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↵1 This work was supported by National Institute for Environmental Health Sciences Grants ES05002 and ES05707 (to I.N.P.) and an American Heart Association, Western States Affiliate, grant (to E.D.B.).
- Abbreviations:
- HCH
- hexachlorocyclohexane
- IP3,inositol-1
- 4,5-trisphosphate
- SR
- sarcoplasmic reticulum
- [Ca2+]i
- intracellular Ca2+concentration
- ER
- endoplasmic reticulum
- RyR
- ryanodine receptor
- RyR2
- cardiac isoform of the ryanodine receptor
- Po
- open probability
- Received August 3, 1998.
- Accepted November 13, 1998.
- The American Society for Pharmacology and Experimental Therapeutics
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