Plant and Cell Physiology, 1994, Vol. 35, No. 5 793-800
© 1994
Characterization of the Ca2+-Dependent Cl– Efflux in Perfused Chara Cells
Department of Life Science, Faculty of Science, Himeji Institute of Technology, Harima Science Park City Hyogo, 678-12 Japan
The mechanism of the Ca2+-dependent Cl– efflux was studied in tonoplast-free cells, in which the intracellular chemical composition can be freely controlled. Tonoplast-free cells were prepared by perfusing the cell interior of internodal cells of Chara corallina with a medium that contained EGTA. The Ca2+-induced Cl– efflux was measured together with the membrane potential during continuous intracellular perfusion. The dependencies of Cl– efflux and the membrane potential on the intracellular Ca2+ or Cl– concentrations were analyzed. When perfusion was started with medium that contained Ca2+ ions, Cl– efflux and membrane depolarization were induced. The amount of Cl– efflux varied considerably among individual cells. The rate of efflux decreased exponentially but a residual efflux remained detectable. The Cl– efflux was induced at concentrations of Ca2+ ions above 1 µM and reached a maximum at 1 mM. By contrast, the membrane depolarization reached a maximum at about 10 µM Ca2+. The rate of Cl– efflux increased linearly with logarithmic increases in the intracellular Cl– concentrations. These findings suggest that more than two kinds of Ca2+-dependent Cl– channel might be present in the plasma membrane.
Addition of ATP or its removal from the perfusion medium did not affect the Ca2+-dependent Cl– efflux. Calmodulin antagonists slightly inhibited the Ca2+-dependent Cl– efflux.
1Present address: Biological Laboratory, Hitotsubashi University, Naka 2-1, Kunitachi, Tokyo, 186 Japan.
(Received February 2, 1994; Accepted April 27, 1994)
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