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Pflügers Archiv

, Volume 419, Issue 6, pp 553–558 | Cite as

Pharmacological block of Ca2+-activated Cl current in rat vascular smooth muscle cells in short-term primary culture

  • A. Baron
  • P. Pacaud
  • G. Loirand
  • C. Mironneau
  • J. Mironneau
Excitable Tissues and Central Nervous Physiology

Abstract

Ca2+-activated Cl currents were studied in isolated cells from rat portal vein smooth muscle in short-term primary culture using the whole-cell patch-clamp technique. Cl currents can be activated separately by Ca2+ release from intracellular stores (in response to external applications of caffeine or noradrenaline) and by Ca2+ influx through voltage-dependent Ca2+ channels. The effects of several Cl channel blockers and of spironolactone (a substance known to reduce internal Ca2+ loading) on both Cl and Ca2+ currents were examined. Diisothiocyanostilbene-2,2′-disulfonic acid (DIDS), anthracene-9-carboxylic acid (9-AC) and diphenylamine-2,2′-dicarboxylic acid (DPC) inhibited the Ca2+-activated Cl current (IC50 values between 16.5 and 306 μM) with no effects on the inward Ca2+ current and on internal Ca2+ loading (tested by measuring the Ca2+-activated K+ current). These results indicate that the inhibition of Cl current by these compounds is due to a direct interaction with the Cl channel. In contrast, spironolactone inhibited both K+ and Cl currents (IC50=7.6 μM) by reducing the amount of Ca2+ located in the internal stores, whereas the Cl current activated by Ca2+ current through T-type Ca2+ channels was unchanged. This preparation and the protocols developed in this study appears to be appropriate for analysis of substances interfering with Cl channels or intracellular Ca2+ stores.

Key words

Ca2+-activated Cl current Cl channels inhibitors Spironolactone Vascular smooth muscle cells 

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Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • A. Baron
    • 1
  • P. Pacaud
    • 1
  • G. Loirand
    • 1
  • C. Mironneau
    • 1
  • J. Mironneau
    • 1
  1. 1.Laboratoire de Physiologie Cellulaire et Pharmacologie MoléculaireINSERM CJF 88-13, Université de Bordeaux IIBordeaux CédexFrance

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