Dynamics of calcium regulation of chloride currents in Xenopus oocytes.
Ca-activated Cl currents are widely expressed in many cell types and play diverse and important physiological roles. The Xenopus oocyte is a good model system for studying the regulation of these currents. We previously showed that inositol 1,4,5-trisphosphate (IP3) injection into Xenopus oocytes rapidly elicits a noninactivating outward Cl current (ICl1-S) followed several minutes later by the development of slow inward (ICl2) and transient outward (ICl1-T) Cl currents. In this paper, we investigate whether these three currents are mediated by the same or different Cl channels. Outward Cl currents were more sensitive to Ca than inward Cl currents, as shown by injection of different amounts of Ca or by Ca influx through a heterologously expressed ligand-gated Ca channel, the ionotropic glutamate receptor iGluR3. These data could be explained by two channels with different Ca affinities or one channel with a higher Ca affinity at depolarized potentials. To distinguish between these possibilities, we determined the anion selectivity of the three currents. The anion selectivity sequences for the three currents were the same (I > Br > Cl), but ICl1-S had an I-to-Cl permeability ratio more than twofold smaller than the other two currents. The different anion selectivities and instantaneous current-voltage relationships were consistent with at least two different channels mediating these currents. However, after consideration of possible errors, the hypothesis that a single type of Cl channel underlies the complex waveforms of the three different macroscopic Ca-activated Cl currents in Xenopus oocytes remains a viable alternative.
Kuruma
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Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322-3030, USA.
Hartzell
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HC
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology*; D004553:Electric Conductivity; D005260:Female; D007476:Ionophores / Q000494:pharmacology; D009865:Oocytes / Q000502:physiology*; D018408:Patch-Clamp Techniques; D017470:Receptors, Glutamate / Q000502:physiology; D013997:Time Factors; D014982:Xenopus laevis / Q000502:physiology*
D018118:Chloride Channels; D007476:Ionophores; D017470:Receptors, Glutamate; D000001:Calcimycin; D002118:Calcium