中华大蟾蜍卵母细胞质膜存在钙依赖性氯离子通道.PDFVIP

生理学报 Acta Physiologica Sinica, October 25, 2006, 58 (5): 471-476 471 研究论文 中华大蟾蜍卵母细胞质膜存在钙依赖性的氯离子通道 * 皮艳俐，马季骅 ，张培华，段晶晶 武汉科技大学医学院心脏电生理研究室，武汉 430080 摘 要：本文采用双微电极电压钳方法研究了中华大蟾蜍卵母细胞内源性电压门控型离子通道的成分及其生理特性。卵母 细胞去极化至-30 mV 及更正电压时，有一持续的电压依赖性外向电流出现。钾离子通道拮抗剂四乙基氯化氨(tetraethy- lammonium chloride, TEA, 10 mmol/L)和4- 氨基吡啶(4-aminopyridine, 4-AP, 10 mmol/L)协同作用时，该电流只能被抑制到最 大电流幅度的(23.4±0.72)% 。但是，上述浓度的TEA 和4-AP 与氯离子通道拮抗剂5- 硝基-2, 3- 苯酚丙胺苯甲酸盐 (5-nitro- 2,3-phenypropylamino benzoate, NPPB, 30 µmol/L) 、无钙Ringer 氏液或钙离子通道拮抗剂维拉帕米(40 µmol/L)协同作用时， 可分别将此外向电流抑制到最大电流幅度的(2.1±0.08)% 、(2.2±0.04)% 和(3.1±0.15)% 。结果表明，中华大蟾蜍卵母细胞质 膜上除有钾离子电流之外，还存在钙依赖性的氯离子电流。 关键词：中华大蟾蜍；卵母细胞；氯离子通道；电压钳 中图分类号：Q424 ；R331.3 + 8 Calcium-dependent chloride channels in plasma membrane of oocytes from toad, Bufo bufo gargarizans * PI Yan-Li, MA Ji-Hua , ZHANG Pei-Hua, DUAN Jing-Jing Cardio-Electrophysiological Research Laboratory, Medical College, Wuhan University of Science and Technology, Wuhan 430080, China Abstract: In this paper, membrane current properties of the fully-grown oocytes from toad, Bufo bufo gargarizans , were studied by using two-microelectrode voltage clamp technique. Axion of adult female toad was destroyed, and then ovarian lobes containing oocytes 2+ in stage I to VI were removed and incubated in Ca -free ND96 solution with collagenase (1.5 mg/ml) for 1 h. Subsequently, the oocytes 2+ were washed in Ca -free ND96 solution for 10 min to completely remove the follicular layer. For the experiments only the oocytes in stage V and VI were selected and used during 1 to 5 d. The membrane was depolarized from a holding potential of -80 mV to +60 mV in 10 mV step. It was found that a sustained outward current was elicited by depolarization. Potassium channel blockers (tetraethylammonium chl