Asymmetrical distribution of Ca-activated Cl channels in Xenopus oocytes.
Xenopus oocytes are a popular model system for studying Ca signaling. They endogenously express two kinds of Ca-activated Cl currents, I(Cl-1), and I(Cl-2). I(Cl-1) is activated by Ca released from internal stores and, with appropriate voltage protocols, by Ca influx. In contrast, I(Cl-2) activation is dependent on Ca influx. We are interested in understanding how these two different Cl channels are activated differently by Ca from different sources. One could hypothesize that these channels are activated differently because they are differentially localized near the corresponding Ca source. As an initial investigation of this hypothesis, we examined the distribution of I(Cl-1) and I(Cl-2) channels in the oocyte. We conclude that both I(Cl-1) and (Cl-2) channels are primarily localized to the animal hemisphere of the oocyte, but that capacitative Ca influx occurs over the entire oocyte membrane. Evidence supporting this view includes the following observations: 1) Injection of IP3 into the animal hemisphere produced larger and faster I(Cl-1) responses than injection into the vegetal hemisphere. 2) Exposure of the animal hemisphere to Cl-free solution almost completely abolished I(Cl-1) produced by IP3-induced release of Ca from internal stores or by capacitative Ca entry. 3) Loose macropatch recording showed that both I(Cl-1) and I(Cl-2) currents were approximately four times and approximately three times, respectively, more dense in the animal than in the vegetal hemisphere. 4) Confocal imaging of oocytes loaded with fluorescent Ca-sensitive dyes showed that the time course of activation of I(Cl-1) corresponded to the appearance of the wave of Ca release at the animal pole. 5) Ca release and Ca influx, although twofold higher in the animal pole, were evident over the entire oocyte.
Machaca
K
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Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322-3030, USA.
Hartzell
H C
HC
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D016764:Cell Polarity; D018118:Chloride Channels / Q000032:analysis / Q000096:biosynthesis / Q000502:physiology*; D002712:Chlorides / Q000378:metabolism / Q000494:pharmacology; D004558:Electric Stimulation; D005260:Female; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology; D007700:Kinetics; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D015398:Signal Transduction; D013997:Time Factors; D014982:Xenopus laevis
D018118:Chloride Channels; D002712:Chlorides; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium