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[目的 ]建立小鼠粗线期精母细胞T型Ca2 + 通道电流膜片箝记录方法 ,并观察拟除虫菊酯类杀虫剂氰戊菊酯的作用。[方法 ]应用膜片箝技术 ,全细胞方式下记录小鼠粗线期精母细胞T型Ca2 + 通道电流 ,并用各种特异的通道阻断剂和激动剂确定电流特性 ,与体细胞T型Ca2 + 通道电流比较 ;通过胞外给药 ,观察氰戊菊酯对T型Ca2 + 通道电流影响。 [结果 ]当以Cs+ 、TEA+ 阻断K+ 电流 ,记录到一 -6 0mV激活 ,-2 0～ -30mV达到最大值的内向电流 ,该电流呈快速激活快速失活。加入Na+通道阻断剂TTX ,对记录的内向电流无影响 ,表明该内向电流不含有Na+ 电流成分 ,是由Ca2 + 通道开放产生。另该电流对L型Ca2 + 通道激动剂BayK 86 44不敏感 ,但对Cd2 + 、Nifedipine两种Ca2 + 通道阻断剂较敏感 ,提示其为T型Ca2 + 通道电流。与体细胞典型T型Ca2 + 通道比较 ,粗线期精母细胞T型Ca2 + 通道有自身特点。研究氰戊菊酯对记录的T型Ca2 + 通道电流的作用显示 :5 0 μM·L-1氰戊菊酯在加入后 3～ 5min开始抑制T型Ca2 + 通道电流 ,于 9～ 10min达到最大抑制作用 ,并有明显的剂量 反应关系。 [结论 ]小鼠粗线期精母细胞主要存在T型Ca2 + 通道 ,而不含有L型Ca2 + 通道 ,但与典型T型Ca2 + 通道电流比较 ,精母细胞的T型Ca2 + 通道电流又?
[Objective] To establish a method for recording the T-type Ca2 + channel current patch clamp of spermatogonia in mice and to observe the effect of the pyrethroid fenvalerate. [Method] Patch clamp technique was used to record the current of T-type Ca2 + channel of spermatogonia in rat spermatids in whole-cell mode. The current characteristics were determined by various channel blockers and agonists, Ca2 + channel current; through extracellular administration, observe the effect of fenvalerate on T-type Ca2 + channel current. [Results] When the current of K + was blocked by Cs + and TEA +, an inward current of -6 0 mV and a maximum of -2 0 ~ -30 mV were recorded, and the current was rapidly inactivated and rapidly inactivated. Addition of Na + channel blocker TTX had no effect on the recorded inward current, indicating that the inward current did not contain Na + current and was generated by Ca 2+ channel opening. In addition, the current was not sensitive to LK Ca2 + channel agonist BayK 86 44, but sensitive to both Ca2 + and Nifedipine Ca2 + channel blockers, suggesting that it is a T-type Ca2 + channel current. Compared with the typical T cell Ca2 + channels of somatic cells, T-type Ca2 + channels of pachytene spermatocytes have their own characteristics. The study of the effect of fenvalerate on the recorded T-type Ca2 + channel currents showed that fenvalerate at 5 0 μM · L-1 inhibited T-type Ca2 + channel currents at 3-5 min after addition, reaching the maximum at 9-10 min Inhibition, and a clear dose-response relationship. [Conclusion] There are mainly T-type Ca2 + channels but not L-type Ca2 + channels in spermatogonia. However, compared with the typical T-type Ca2 + channel currents, the T-type Ca2 + channels currents of spermatocytes ?