Am J Physiol Cell Physiol AJP: Endocrinology and Metabolism
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Am J Physiol Cell Physiol 274: C161-C166, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 1, C161-C166, January 1998

Differences in regulation of Ca2+-activated Clminus channels in colonic and parotid secretory cells

Jorge Arreola1,2, James E. Melvin1, and Ted Begenisich2

Departments of 1 Dental Research and of 2 Pharmacology and Physiology, University of Rochester Medical Center, Rochester, New York 14642

We investigated the regulation of Ca2+-activated Cl- channels in cells from the human colonic cell line T84 and acinar cells from rat parotid glands. The participation of multifunctional Ca2+- and calmodulin-dependent protein kinase (CaM kinase) II in the activation of these channels was studied using selective inhibitors of calmodulin and CaM kinase II. Ca2+-dependent Cl- currents were recorded using the whole cell patch-clamp technique. Direct inhibition of CaM kinase II by 40 µM peptide 281-302 or by 10 µM KN-62, another CaM kinase inhibitor, did not block the Cl- current in parotid acinar cells, whereas in T84 cells KN-62 markedly inhibited the Ca2+-dependent Cl- current. We also used the calmodulin-binding domain peptide 290-309 (0.5 µM), which competitively inhibits the activation of CaM kinase II. This peptide reduced the Cl- current in T84 cells by ~70% but was without effect on the channels in parotid acinar cells. We conclude that the Ca2+-dependent Cl- channels in T84 cells are activated by CaM kinase II but that the channels in parotid acinar cells must be regulated by a fundamentally different Ca2+-dependent mechanism that does not utilize CaM kinase II or any calmodulin-dependent process.

exocrine acinar cells; human colon carcinoma cells; fluid and electrolyte secretion; calmodulin; calmodulin and calmodulin kinase inhibitors


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