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13.
Role of sarcoplasmic reticulum and mitochondria in ▫$ca^2+$▫ removal in airway myocytes
Roux, Etienne ; Marhl, Marko
The aim of this was to use both a theoretical and experimental approach to determine the influence of the sarco-endoplasmic ▫$Ca^2+$▫ -ATPase (SERCA) activity and mitochondria ▫$Ca^2+$▫ uptake on ...▫$Ca^2+$▫ homeostasis in airway myocytes. Experimental studies were performed on myocytes freshly isolated from rat trachea. ▫$[Ca^2+]_i$▫ was measured by microspectrofluorimetry using indo-1. Stimulation by caffeine for 30 s induced a concentration-graded response characterized by a transient peak followed by a progressive decay to a plateau phase. The decay phase was accelerated for 1-s stimulation, indicating ryanodine receptor closure. In ▫$Na^2+ -Ca^2+$▫ -free medium containing 0.5mM ▫$La^3+$▫,the ▫$[Ca^2+]_i$▫ response pattern was not modified, indicating no involvement of transplasmalemmal ▫$Ca^2+$▫ fluxes. The mathematical model describing the mechanism of ▫$Ca^2+$▫ handling upon RyR stimulation predicts that after ▫$Ca^2+$▫ release from the sarcoplasmic reticulum, the ▫$Ca^2+$▫ is first sequestrated by cytosolic proteins and mitochondria, and pumped back into the sarcoplasmic reticulum after a time delay. Experimentally, we showed that the ▫$[Ca^2+]_i$▫ decay after increase was not altered by the SERCA inhibitor cyclopiazonic acid, but was slightly but significantly modified by the mitochondria uncoupler carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone. The experimental and theoretical results indicate that, although ▫$Ca^2+$▫ pumping back by SERCA is active, it is not primarily involved in ▫$[Ca^2+]_i$▫ decrease that is due, in part, to mitochondrial ▫$Ca^2+$▫ uptake
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