We investigated the effects of internal pH on Ca-activated, nucleotide-inhibited nonselective cation channels in the basolateral membranes of mouse collecting tubules, using the inside-out variant of the patch clamp technique. pH modulated the channel open probability (P o ), giving a bell-shaped curve peaking at pH 6.8/7.0: P o at pH 6.0 was 11±6% of P o at pH 7.2 and 32 ±7% at pH 8.0. The open and closed time distributions, best fitted to the sum of two exponentials, were differently sensitive to acid and alkaline conditions. Low pH reduced the short and long open times to 38 and 24% of their pH 7.2 values, while high pH produced a 4-fold increase in the long closed time. As previously reported, 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid (SITS) induced a quasi-permanent opening of the channel. The inhibition of the channel produced by high pH disappeared in the presence of SITS, while the inhibition produced by low pH was unaffected. These results suggest that the pH dependence of the channel is due to two separate mechanisms. pH was without effect on the ATP-evoked inhibition of the channel, while high pH profoundly reduced the steepness of the AMP inhibition curve, without altering the half-maximal inhibitory AMP concentration.
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We thank M. Blonde for expert technical assistance, and M. Poitou for secretarial help. A. Chraïbi was supported by the French Society of Nephrology, R. Guinamard is the recipient of a MRE research studentship and J. Teulon is a CNRS researcher. The manuscript was edited by Owen Parkes.
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Chraïbi, A., Guinamard, R. & Teulon, J. Effects of internal ph on the nonselective cation channel from the mouse collecting tubule. J. Membarin Biol. 148, 83–90 (1995). https://doi.org/10.1007/BF00234159
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