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Related Experiment Videos

Ca2+ increase and pH decrease induced by hypochlorous acid in single quiescent myocytes isolated from rat ventricles

M Kuroda1, T Kaminishi, K Uchida

  • 11st Department of Internal Medicine, Yamagata University School of Medicine, Japan.

The Japanese Journal of Physiology
|January 1, 1995
PubMed
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Hypochlorous acid (HOCl) affects heart cells by altering calcium levels and acidity. This study shows HOCl increases cytosolic free calcium concentration and decreases intracellular pH, with changes linked to intracellular stores.

Area of Science:

  • Cardiovascular Physiology
  • Cellular Electrophysiology

Background:

  • Cytosolic free calcium concentration ([Ca2+]i) and intracellular pH (pHi) are critical for cardiac myocyte function.
  • Hypochlorous acid (HOCl) is a reactive oxygen species with potential effects on cellular homeostasis.

Purpose of the Study:

  • To investigate the simultaneous effects of HOCl on [Ca2+]i and pHi in isolated rat ventricular myocytes.
  • To elucidate the source of calcium changes and the mechanism of HOCl-induced alterations.

Main Methods:

  • Simultaneous measurement of [Ca2+]i and pHi using fluorescence techniques (fura-2 and BCECF).
  • Treatment of myocytes with HOCl and assessment of changes in [Ca2+]i and pHi.
  • Investigation of calcium sources using extracellular calcium chelation and calcium antagonists.

Related Experiment Videos

  • Use of a disulfide-reducing agent (1,4-dithiothreitol, DTT) to assess reversibility.

    • Main Results:

    • HOCl (200 microM) significantly increased [Ca2+]i by 129 +/- 18nM and decreased pHi by 0.18 +/- 0.02.
    • A positive linear correlation was observed between changes in [Ca2+]i and pHi.
    • The increase in [Ca2+]i was primarily attributed to intracellular calcium stores, not extracellular influx.
    • DTT treatment restored both [Ca2+]i and pHi to baseline levels.

    Conclusions:

    • HOCl induces simultaneous alterations in cytosolic calcium and intracellular pH in cardiac myocytes.
    • The observed changes are mainly driven by intracellular calcium release and are reversible with a reducing agent.
    • These findings highlight the impact of HOCl on cardiac cell ion homeostasis.