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Previous studies have shown that the V(j)-dependent gating behavior of gap junction channels is altered by CO(2) exposure. V(j)-dependent channel closure is increased by CO(2) in some connexin channels and decreased in others. Since the former type of channels gate on the relatively negative side by V(j) (negative gaters) and the latter at the positive side (positive gaters), it has been hypothesized that gating polarity determines the way CO(2) affects V(j) closure. To test this hypothesis, we have studied the CO(2)-mediated changes in V(j) gating in channels made of Cx32, Cx26, or a Cx32 mutant (Cx32-N2D) in which asparagine (N) at position 2 was replaced with aspartate (D). With exposure to CO(2), Cx32 channels (negative gaters) show increased V(j)-dependent closure, whereas Cx26 channels (positive gaters) respond in the opposite way to V(j). Additionally, Cx32-N2D channels (positive gaters) show decreased V(j) closure with exposure to CO(2). The reciprocal Cx26 mutant, Cx26-D2N (negative gater), could not be tested because it did not express functional homotypic channels. The data support the hypothesis that polarity of fast V(j) gating determines whether CO(2) increases or decreases the V(j) dependent closure of gap junction channels.
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