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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。
OBJECTIVE: To study the cell viability of vestibular hair cells and the type of potassium channel in the membrane. METHODS: Guinea pig balloon hair cells were isolated by mechanical digestion after enzyme digestion. Potassium channel currents were observed on the membrane of guinea pig type Ⅱ hair cells by whole cell patch clamp technique. RESULTS: ① When the collagenase Ⅳ concentration was 0 35mg / ml, the number of isolated hair cells was the most, and the survival time was the longest. ② When the clamp potential was-10 0mV, with a step of 10mV, from 70mV to + 20mV With the depolarization of the membrane potential, a series of rapid and transient outward currents, mainly type A potassium channels, were recorded, and 4 Ap specifically blocked it. The activation voltage is -60 ~ -50mV. When the clamp potential is -70mV, step-depolarization (-500mV ~ + 40mV) with 10mV steps can produce a series of delayed rectifier potassium currents, TEA This current can be reduced by 5 3 3% ± 6 0%. Conclusion: The optimal concentration of hair follicle enzyme isolated from guinea pig balloon is 0 35mg / ml. Type A hair cell membrane has type A potassium channel and delayed rectifier potassium channel.