| Abstract |
1. An outwardly rectifying Cl- (ORCl) current of murine osteoclasts was activated by hypotonic stimulation. The current was characterized by rapid activation, little inactivation, strong outward rectification, blockage by DIDS and permeability to organic acids (pyruvate and glutamate). 2. The hypotonically activated ORCl current was inhibited by intracellular dialysis with an ATP-free pipette solution, but not by replacement of ATP with a poorly hydrolysable ATP analogue adenosine 5'-O-(3-thiotriphosphate). The current amplitude was reduced when intracellular alkalinity increased over the pH range 6.6-8.0. 3. Intracellular application of cytochalasin D occasionally activated the ORCl current without hypotonic stress, but inhibited activation of the ORCl current by hypotonic stimulation. The hypotonically activated ORCl current was unaffected by a non-actin-depolymerizing cytochalasin, chaetoglobosin C, but partially inhibited by deoxyribonuclease I. 4. Removal of extracellular Ca2+ inhibited activation of the ORCl current by hypotonic shock, but did not reduce the current once activated. The hypotonically activated ORCl current was partially decreased by intracellular dialysis with 20 mM EGTA. 5. With 10 mM Ca2+ in the extracellular medium, the ORCl current was activated in response to more minor decreases in osmolarity than with 1 mM Ca2+. The increased sensitivity to hypotonicity was mimicked by increasing the intracellular Ca2+ level (pCa 6.5). 6. These results suggest that hypotonic stimulation and a rise in the extracellular Ca2+ level synergistically activate the ORCl channel of murine osteoclasts, and that the activating process is modified by multiple intracellular factors (pH, ATP and actin cytoskeletal organization).
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