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Ca2+-activated Cl channels in Ehrlich ascites tumor cells are distinct from mCLCA1, 2 and3

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Using the whole-cell patch-clamp technique, we have studied the electrophysiological and pharmacological properties of the Ca2+-activated Cl current present in Ehrlich cells. The currents activated slowly upon depolarization, deactivated upon hyperpolarization, and showed strong outward rectification. An increase in [Ca2+]i activated the current with an EC50 of 165.2 nM. Extracellular application of niflumic acid (100 µM) rapidly blocked the current in a voltage-dependent manner whereas sulfhydryl-modifying agents such as dithiothreitol (DTT, 1–2 mM) and N-ethylmaleimide (NEM, 100 µM) had no effect on Ca2+-activated currents in Ehrlich cells. Members of the recently discovered CLCA gene family are the only molecular candidates for Ca2+-activated Cl channels cloned so far. Using RT-PCR we demonstrated that the appearance of a Ca2+-activated Cl current in Ehrlich cells is not associated with the expression of the murine members of the CLCA family (mCLCA1–mCLCA3). Correspondingly, the kinetic and pharmacological properties of the Ca2+-activated current in Ehrlich cells differ from those of CLCA-associated currents, which are time independent and DTT sensitive. Thus, phenotypic differences in combination with RT-PCR data point to the existence of different molecular species for Ca2+-activated Cl channels.

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Received after revision: 15 December 2000

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Papassotiriou, .J., Eggermont, .J., Droogmans, .G. et al. Ca2+-activated Cl channels in Ehrlich ascites tumor cells are distinct from mCLCA1, 2 and3. Pflügers Arch - Eur J Physiol 442, 273–279 (2001).

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