Swelling-activated Cl- currents, I(ci,swell), were measured during hyposmotic shock in white Leghorn embryonic chick heart cells using the whole-cell recording of patch-clamp technique. Genistein, an inhibitor of prot...Swelling-activated Cl- currents, I(ci,swell), were measured during hyposmotic shock in white Leghorn embryonic chick heart cells using the whole-cell recording of patch-clamp technique. Genistein, an inhibitor of protein tyrosine kinase (PTK), suppressed I(ci,swell), Under isosmotic condition phorbol 12-myristate 13-acetate (PMA), an activator of PKC, elicited the Cl~ current similar to that in hyposmotic solution, whereas hyposmotic shock did not elicit I(ci,swell) in chelerythrine chloride(an inhibitor of PKC)-treated cells. Con-focal microscopy experiments using FITC-phalloidin as a fluorescent label of F-actin showed that the actin network was moved from cortical region of the cell to the center after hyposmotic shock as compared with the image under isosmotic condition. When the cells were treated with cytochalasin B (CB) or cytochalasin D (CD) under isosmotic condition the disruption of the F-actin integrity was observed, and I(ci,Sweii) was not elicited. With combination treatment of CB with PMA, hyposmotic solution could not elicited I(Ci,swell), The results suggested that the role of PTK, probably receptor tyrosine kinase, for regulation of I(ci,sweii) appeared to be at upstream site related to the role of F-actin. Then PKC signal pathway was activated somehow and finally change in the polymerization state of cytoskeleton led to activate the swelling-activated Cl- channels. These results demonstrate clearly that PTK, PKC and F-actin are important factors for regulation of I(Ci,swell) in embryonic chick heart cells as compared with often controversial results reported in different cell types.展开更多
基金This work was supported by National Natural Science Foundation of China (No. 30070205 and No. 39730150).
文摘Swelling-activated Cl- currents, I(ci,swell), were measured during hyposmotic shock in white Leghorn embryonic chick heart cells using the whole-cell recording of patch-clamp technique. Genistein, an inhibitor of protein tyrosine kinase (PTK), suppressed I(ci,swell), Under isosmotic condition phorbol 12-myristate 13-acetate (PMA), an activator of PKC, elicited the Cl~ current similar to that in hyposmotic solution, whereas hyposmotic shock did not elicit I(ci,swell) in chelerythrine chloride(an inhibitor of PKC)-treated cells. Con-focal microscopy experiments using FITC-phalloidin as a fluorescent label of F-actin showed that the actin network was moved from cortical region of the cell to the center after hyposmotic shock as compared with the image under isosmotic condition. When the cells were treated with cytochalasin B (CB) or cytochalasin D (CD) under isosmotic condition the disruption of the F-actin integrity was observed, and I(ci,Sweii) was not elicited. With combination treatment of CB with PMA, hyposmotic solution could not elicited I(Ci,swell), The results suggested that the role of PTK, probably receptor tyrosine kinase, for regulation of I(ci,sweii) appeared to be at upstream site related to the role of F-actin. Then PKC signal pathway was activated somehow and finally change in the polymerization state of cytoskeleton led to activate the swelling-activated Cl- channels. These results demonstrate clearly that PTK, PKC and F-actin are important factors for regulation of I(Ci,swell) in embryonic chick heart cells as compared with often controversial results reported in different cell types.
