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Attenuation of stimulated Ca2+ influx in colonie epithelial (HT29) cells by cAMP

  • Original Article
  • Molecular and Cellular Physiology
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Abstract

In HT29 colonie epithelial cells agonists such as carbachol (CCH) or ATP increase cytosolic Ca2+ activity ([Ca2+]i) in a biphasic manner. The first phase is caused by inositol 1,4,5-trisphophate- (Insp 3−) mediated Ca2+ release from their respective stores and the second plateau phase is mainly due to stimulated transmembraneous Ca2+ influx. The present study was undertaken to examine the effect of increased adenosine 3′,5′-cyclic monophasphate (cAMP) (forskolin 10 μmol/1 = FOR) on the Ca2+ transient in the presence of CCH (100 μmol/l). In unpaired experiments it was found that FOR induced a depolarization and reduced cytosolic Ca2+ ([Ca2+]i, measured as the fura-2 fluorescence ratio 340/380 nm) significantly. Dideoxyforskolin had no such effect. The effect of FOR was abolished when the cells were depolarized by a high-K+ solution. In further paired experiments utilizing video imaging in conjunction with whole-cell patch-clamp, [Ca2+]i was monitored separately for the patch-clamped cell and three to seven neighbouring cells. In the presence of CCH, FOR reduced [Ca2+]i uniformly from a fluorescence ratio (345/380) of 2.9 ± 0.12 to 1.8 ± 0.07 in the patch-clamped cell and its neighbours (n = 48) and depolarized the membrane voltage (V m) of the patch-clamped cells significantly and reversibly from − 54 ± 7.4 to − 27 ± 5.9 mV (n = 6). In additional experiments V m was depolarized by 15–54 mV by various increments in the bath K+ concentration. This led to corresponding reductions in [Ca2+]i. Irrespective of the cause of depolarization (high K+ or FOR) there was a significant correlation between the change in V m and change in [Ca2+]i. These data indicate that the cAMP-mediated attenuation of Ca2+ influx is caused by the depolarization produced by this second messenger.

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Authors and Affiliations

  1. Physiologisches Institut der Albert-Ludwigs-Universität, Hermann-Herder-Strasse 7, D-79104, Freiburg, Germany

    J. Leipziger, P. Rubini-Illes, R. Nitschke & R. Greger

  2. Abteilung IV Nephrologie, Medizinische Klinik, Albert-Ludwigs-Universität, Hugstetter Strasse 55, D-79106, Freiburg, Germany

    K. -G. Fischer

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  1. K. -G. Fischer
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  2. J. Leipziger
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  3. P. Rubini-Illes
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  4. R. Nitschke
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  5. R. Greger
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Fischer, K.G., Leipziger, J., Rubini-Illes, P. et al. Attenuation of stimulated Ca2+ influx in colonie epithelial (HT29) cells by cAMP. Pflügers Arch. 432, 735–740 (1996). https://doi.org/10.1007/s004240050192

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  • DOI: https://doi.org/10.1007/s004240050192

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