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Pharmacological Preconditioning Using Diazoxide Regulates Store-Operated Ca2 + Channels in Adult Rat Cardiomyocytes.


ABSTRACT: Voltage-dependent Ca2+ channels and store-operated Ca2+ channels (SOCs) are the major routes of Ca2+ entry into mammalian cells. Previously, we reported that pharmacological preconditioning (PPC) leads to a decrease in the amplitude of L-type calcium channel current in the heart. In this study, we examined PPC-associated changes in SOC function. We measured adult cardiomyocyte membrane currents using the whole-cell patch-clamp technique, and we evaluated reactive oxygen species (ROS) production and intracellular Ca2+ levels in cardiomyocytes using fluorescent probes. Diazoxide (Dzx) and thapsigargin (Tg) were used to induce PPC and to deplete internal stores of Ca2+, respectively. Ca2+ store depletion generated inward currents with strong rectification, which were suppressed by the SOC blocker GSK-7975-A. These currents were completely abolished by PPC, an effect that could be countered with 5-hydroxydecanoate (5-HD; a selective mitochondrial ATP-sensitive K+ channel blocker), an intracellular mitochondrial energizing solution, or Ni2+ [a blocker of sodium-calcium exchanger (NCX)]. Buffering of ROS and intracellular Ca2+ also prevented PPC effects on SOC currents. Refilling of intracellular stores was largely suppressed by PPC, as determined by measuring intracellular Ca2+ with a fluorescent Ca2+ indicator. These results indicate that influx of Ca2+ through SOCs is inhibited by their ROS and Ca2+-dependent inactivation during PPC and that NCX is a likely source of PPC-inactivating Ca2+. We further showed that NCX associates with Orai1. Down-regulation of SOCs by PPC may play a role in cardioprotection following ischemia-reperfusion.

SUBMITTER: Sampieri R 

PROVIDER: S-EPMC6972595 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Pharmacological Preconditioning Using Diazoxide Regulates Store-Operated Ca<sup>2 +</sup> Channels in Adult Rat Cardiomyocytes.

Sampieri Raúl R   Fuentes Eridani E   Carrillo Elba D ED   Hernández Ascención A   García María C MC   Sánchez Jorge A JA  

Frontiers in physiology 20200114


Voltage-dependent Ca<sup>2+</sup> channels and store-operated Ca<sup>2+</sup> channels (SOCs) are the major routes of Ca<sup>2+</sup> entry into mammalian cells. Previously, we reported that pharmacological preconditioning (PPC) leads to a decrease in the amplitude of L-type calcium channel current in the heart. In this study, we examined PPC-associated changes in SOC function. We measured adult cardiomyocyte membrane currents using the whole-cell patch-clamp technique, and we evaluated reactive  ...[more]

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