拟环纹豹蛛神经细胞的急性分离及其电压门控钠通道的膜片钳研究

对拟环纹豹蛛雌蛛中枢神经细胞进行急性分离,并利用全细胞膜片钳技术对神经细胞的电压门控钠通道基本电生理学特性进行了研究,同时测定了氯氰菊酯及其不同浓度对Na 通道门控过程的影响。结果表明,拟环纹豹蛛头胸部神经团在消化酶和培养液混合作用下能急性分离出单个神经细胞,在4～6h内适合用于膜片钳研究。在全细胞电压钳条件下,神经细胞Na 通道电流(INa)在-60mV左右激活, 10mV左右达峰值,对河豚毒素敏感。用氯氰菊酯处理后,Na 通道的激活电压向负电位方向移动约10mV,为-70mV左右,峰值提前在 5mV左右出现。INa先增大然后迅速减小。用不同浓度的氯氰菊酯处理神经细胞后,INa在各电压下的差异不大,但达到最大INa所需时间随氯氰菊酯浓度的增加而缩短。

Acute isolation of nerve cells from Pardosa pseudoannulata

The nerve cells were isolated acutely from the ventral nerve cord of the female spider of Pardosa pseudoannulata.The properties of the voltage-gated sodium ion channels of the acute separated neurons and the effects of cypermethrin on voltage-gated sodium ion channels were studied with whole-cell patch-clamp technique.The separated neurons were suitable to be used in the patch-clamp experimental research,but best be used in 4-6 hours after being isolated.Under whole-cell patch-clamp recording,sodium current(INa)was activated when the voltage was around-60 mV,and the peak current occurred at about 10 mV.INa was sensitive to tetrodotoxin.After treatment for the nerve cell with cypermethrin,the activated voltage of sodium channel shifted towards more negative(the value was around-70 mV)and the peak current occurred at about 5 mV.INa increased at the beginning,and then reduced rapidly.When nerve cells were treated with cypermethrin at different concentrations,there was no significant difference among INa at different voltages,but the duration to peak current became shorter with the increasing of cypermethrin concentrations.