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目的:建立爪蟾卵母细胞双电极电压钳记录技术。方法:在去除滤泡膜的非洲爪蟾卵母细胞上,建立双电极电压钳记录技术,并使用该技术观察爪蟾卵母细胞表达的内源性递质门控性和电压门控性离子通道电流。结果:灌流液中分别加入NMDA受体、γ-氨基丁酸和烟碱受体激动剂,均未能在爪蟾卵母细胞上诱发出电流。去极化刺激仅可诱发出一外向电流。此电流的幅度随细胞外液中Ca2+浓度的增加而增大。当外液中无Ca2+或无C l-时,电流消失。此电流不被TEA所阻断,但能被MnC l2可逆性阻断。结论:成功建立了爪蟾卵母细胞双电极电压钳记录技术。去极化刺激在爪蟾卵母细胞上诱发出的外向电流为钙依赖性C l-电流。
Objective: To establish a two-electrode voltage clamp recording technology of Xenopus oocytes. Methods: Two-electrode voltage-clamp recording technique was established on Xenopus laevis oocytes with follicular membrane exclusion. The technique was used to observe the endogenous neurotransmitter-gated and voltage-gated ions expressed in Xenopus oocytes Channel current. Results: NMDA receptor, γ-aminobutyric acid and nicotinic receptor agonist were respectively added into the perfusate fluid, which failed to induce current on Xenopus oocytes. Depolarization stimulation can only induce an outward current. The magnitude of this current increases with increasing Ca2 + concentration in extracellular fluid. When the external solution without Ca2 + or without C l-, the current disappears. This current is not blocked by TEA, but is reversibly blocked by MnC12. Conclusion: Two-electrode voltage-clamp recording technique of Xenopus oocytes has been successfully established. Depolarization stimulated the outward current induced on Xenopus oocytes to be a Ca-dependent Cl-current.