Skip to main content
Log in

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

  • Published:
Pflügers Archiv Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Amédée T, Large WA, Wang Q (1990) Characteristics of chloride currents activated by noradrenaline in rabbit ear artery cells. J Physiol (Lond) 428:501–516

    Google Scholar 

  • Bolton TB, Lim SP (1989) Properties of calcium stores and transient outward currents in single smooth muscle cells of rabbit intestine. J Physiol (Lond) 409:385–401

    Google Scholar 

  • Bretag AH (1987) Muscle chloride channels. Physiol Rev 67:618–724

    Google Scholar 

  • Byrne NG, Large WA (1987) Action of noradrenaline on single smooth muscle cells freshly dispersed from the rat anococcygeus muscle. J Physiol (Lond) 389:513–525

    Google Scholar 

  • Byrne NG, Large WA (1988) Membrane ionic mechanisms activated by noradrenaline in cells isolated from the rabbit portal vein. J Physiol (Lond) 404:557–573

    Google Scholar 

  • Dacquet C, Loirand G, Mironneau C, Mironneau J, Pacaud P (1987) Spironolactone inhibition of contraction and calcium channels in rat portal vein. Br J Pharmacol 92:535–544

    Google Scholar 

  • Droogmans G, Casteels R (1981) Temperature-dependence of 45Ca fluxes and contraction in vascular smooth muscle cells of rabbit ear artery. Pflügers Arch 391:183–189

    Google Scholar 

  • Franciolini F, Petris A (1990) Chloride channels of biological membranes. Biochim Biophys Acta 1024:247–25

    Google Scholar 

  • Gögelein H (1988) Chloride channels in epithelia. Biochim Biophys Acta 947:521–547

    Google Scholar 

  • Hamill OP, Marty A, Neher E, Sakmann E, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membranes patches. Pflügers Arch 391:85–100

    Google Scholar 

  • Hughes AD, Hering S, Bolton TB (1990) The action of caffeine on inward barium current through voltage-dependent calcium channels in single rabbit ear artery cells. Pflügers Arch 416:462–466

    Google Scholar 

  • Landry DW, Akabas MH, Redhead C, Edelman A, Cragoe EJ, Al Awqati Q (1989) Purification and reconstitution of chloride channels from kidney and trachea. Science 244:1469–1472

    Google Scholar 

  • Loirand G, Pacaud P, Mironneau C, Mironneau J (1986) Evidence for two distinct calcium channels in rat vascular smooth muscle cells in short-term primary culture. Pflügers Arch 407:566–568

    Google Scholar 

  • Martin C, Dacquet C, Mironneau C, Mironneau J (1989) Caffeine-induced inhibition of calcium channel current in cultured smooth muscle cells from pregnant rat myometrium. Br J Pharmacol 98:493–498

    Google Scholar 

  • Missiaen L, Declerck I, Droogmans G, Plessers L, De Smedt H, Raeymaekers L, Casteels R (1990) Agonist-dependent Ca2+ and Mn2+ entry dependent on state of filling of Ca2+stores in aortic smooth muscle cells of the rat. J Physiol (Lond) 427:171–186

    Google Scholar 

  • Muallem S, Khademazad M, Sachs G (1990) The route of Ca2+ entry during reloading of the intracellular Ca2+ pool in pancreatic acini. J Biol Chem 265:2011–2016

    Google Scholar 

  • Pacaud P, Loirand G, Lavie JL, Mironneau C, Mironneau J (1989a) Calcium-activated chloride current in rat vascular smooth muscle cells in short-term primary culture. Pflügers Arch 413:629–636

    Google Scholar 

  • Pacaud P, Loirand G, Mironneau C, Mironneau J (1989b) Noradrenaline activates a calcium-activated chloride conductance and increases the voltage-dependent calcium current in cultured single cells of rat portal vein. Br J Pharmacol 97:139–146

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Baron, A., Pacaud, P., Loirand, G. et al. Pharmacological block of Ca2+-activated Cl current in rat vascular smooth muscle cells in short-term primary culture. Pflügers Arch 419, 553–558 (1991). https://doi.org/10.1007/BF00370294

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00370294

Key words

Navigation