豚鼠球囊毛细胞的分离及钾离子流的研究  

THE STUDIES OF CELL ISOLATION AND POTASSIUM CURRENTS IN GUINEA-PIG SACCULAR HAIR CELLS

作　　者：沙建慧[1] 李云义[1] 李云阁[1] 刘焕珠[1] 王清 姚萍[1] 

机构地区：[1]第四军医大学吉林军医学院生理教研室,吉林市132013

出　　处：《中国应用生理学杂志》2000年第3期279-282,共4页Chinese Journal of Applied Physiology

摘　　要：目的 :研究前庭毛细胞的细胞活性及膜上钾通道的类型。方法 :用酶消化后机械法分离豚鼠球囊毛细胞 ,并用全细胞膜片钳技术观察豚鼠球囊Ⅱ型毛细胞侧膜上的钾通道电流。结果 :①胶原酶Ⅳ浓度为 0 35mg/ml时 ,分离的毛细胞数量最多 ,存活时间最长 ;②当钳制电位为 - 10 0mV ,以 10mV的步距 ,从 - 70mV至 + 2 0mV阶跃 ,随着膜电位的去极化 ,可记录到一系列快速、瞬时的以A型钾通道为主的外向电流 ,4 Ap对其有特异性阻断作用。激活电压为 - 6 0～ - 5 0mV ;③当钳制电位为 - 70mV ,以 10mV的步距阶跃去极化 (- 5 0mV～ + 40mV) ,可产生一系列延迟整流性钾离子流 ,TEA能使该电流幅度下降 5 3 3 %± 6 0 %。结论 :分离豚鼠球囊毛细胞酶的最佳浓度为 0 35mg/ml,Ⅱ型毛细胞膜上有A型钾通道和延迟整流性钾通道。Aim:To study the cell viability in the guinea-pig's saccular hair cells and the type of potassium channels on their membrane.Methods:Enzymatic and mechanical dissociation were used to separate guinea-pig's saccular hair cells and whole cell patch-clamp technique was used to observe potassium current on guinea-pig's saccular Ⅱ type hair cells membrane. Results:①When the concentration of collagenase Ⅳ was 0.35 mg/ml,single living hair cells were the most numerous and the longest in survial.②A series of rapid and instant outward currents of which were A-type of potassium channel mainly could be recorded when the holding potential was at -100mV and the depolarized potentials from -70 mV to +20 mV were in 10 mV increments.They could be blocked by 4-Ap especially.The activated membrane potential was (-60～-50)mV.③When the holding potential was at -70 mV and the depolarized potentials from -50 mV to +40 mV were in 10 mV increments,depolariztion could produce a series of delayed rectifier potassium currents .TEA could decrease it by 53.3±6.0%.Conclusion: The enzymatic concentration of 0.35 mg/ml could achieve the best result in isolating saccular hair cells of guinea-pig.There are A-type and delaged rectifier potassium channel on guinea-pig's saccular Ⅱ type hair cells' membrane.

关 键 词：球囊 毛细胞 K+电流 膜片钳技术 

分 类 号：R339.174[医药卫生—人体生理学]