卵母细胞双电极电压钳技术的建立及其记录到的内源性外向电流

Establishment of two-microelectrode voltage-clamp technique for oocytes and the recorded endogenous outward currents

目的:建立卵母细胞双电极电压钳记录技术。方法:建立双电极电压钳技术平台,并应用此平台,在去除滤泡膜的中国四川牛蛙卵母细胞上,记录内源性的外向离子电流。结果:卵母细胞孵育在正常ND96溶液中,将细胞膜钳制在-80 mV膜电位,给予一系列的去极化刺激可诱发出一个外向电流。在细胞外液中加入10 mM的TEA可阻断该电流80%以上,洗脱后电流幅值恢复至90%左右。将正常ND96溶液替换为无钙ND96溶液后,电流没有变化。结论:成功建立了卵母细胞双电极电压钳记录技术。去极化刺激在牛蛙卵母细胞上诱发出的外向电流为钾电流。

Objective： To establish two-microelectrode voltage-clamp technique for oocytes. Methods： Two-microelectrode voltage-clamp technique platform was established and the endogenous outward ionic currents were recorded on bullfrog oocytes after the removal of follicular theca. Results： As the oocytes were incubated in normal ND96 solution, the membrane potential was maintained at -80 mV, then depolarized to ＋50 mV in 10 mV increments, the outward cuttents were recorded. After applied 10 mmol/L TEA in the extracellular fluid, more than 80% currents were blocked, then recovery to about 90% after washout. The currents did not obviously change in ND96 solution without calcium. Conclusion： Two -microelectrode voltage-clamp technique is successfully established in oocytes. The outward currents induced by depolarizing stimulation on bullfrog oocytes were potassium currents.