Simultaneous analysis of cell Ca2+ and Ca2(+)-stimulated chloride conductance in colonic epithelial cells (HT-29)

Cell Regulation
A P MorrisR A Frizzell

Abstract

We used perforated patch, whole-cell current recordings and video-based fluorescence ratio imaging to monitor the relation of plasma membrane ionic conductances to intracellular free Ca2+ within individual colonic epithelial cells (HT-29). The Ca2(+)-mediated agonist, neurotensin, activated K+ and Cl- conductances that showed different sensitivities to [Ca2+]i. The Cl- conductance was sensitive to increases or decreases in [Ca2+]i around the resting value of 76 +/- 32 (mean +/- SD) nM (n = 46), whereas activation of the K+ conductance required at least a 10-fold rise in [Ca2+]i. Neurotensin increased [Ca2+]i by stimulating a transient intracellular Ca2+ release, which was followed by a sustained rise in [Ca2+]i due to Ca2+ influx from the bath. The onset of the initial [Ca2+]i transient, monitored at a measurement window over the cell interior, lagged behind the rise in Cl- current during agonist stimulation. This lag was not present when the [Ca2+]i rise was due to Ca2+ entry from the bath, induced either by the agonist or by the Ca2+ ionophore ionomycin. The temporal differences in [Ca2+]i and Cl- current during the agonist-induced [Ca2+]i transient can be explained by a localized Ca2+ release from intracellular stores in the...Continue Reading

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Citations

Sep 1, 1991·Pflügers Archiv : European journal of physiology·M BöhmeW Rummel
Oct 1, 1994·Pflügers Archiv : European journal of physiology·A W CuthbertM J Evans
Sep 1, 1992·Pflügers Archiv : European journal of physiology·M BidetP Poujeol
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May 14, 2005·Annals of the New York Academy of Sciences·Estelle LouisetHubert Vaudry
Sep 6, 2012·British Journal of Pharmacology·Yuemin TianKarl Kunzelmann
Jun 9, 2000·Annual Review of Physiology·J F Kidd, P Thorn
Apr 2, 2005·Viral Immunology·Judith M BallRebecca D Parr
Nov 14, 1997·The American Journal of Physiology·L GamaG E Breitwieser
Aug 4, 1998·The American Journal of Physiology·D MerlinJ L Madara
Aug 13, 1999·The American Journal of Physiology·A P MorrisM K Estes
Sep 1, 1993·The American Journal of Physiology·W M Moran, J D Valentich
Apr 1, 1991·The American Journal of Physiology·R T Worrell, R A Frizzell
Apr 1, 1993·The American Journal of Physiology·A P Morris, R A Frizzell

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