REPLATING IMPROVES WHOLE CELL VOLTAGE CLAMP RECORDING OF HUMAN FETAL DORSAL-ROOT GANGLION NEURONS
REPLATING IMPROVES WHOLE CELL VOLTAGE CLAMP RECORDING OF HUMAN FETAL DORSAL-ROOT GANGLION NEURONS
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DOI：
10.1016/0165-0270(90)90094-v
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发表时间：
1990-10-01
影响因子：
3
通讯作者：
RAPOPORT, SI
中科院分区：
文献类型：
--
作者：
CAVIEDES, P;AULT, B;RAPOPORT, SI
整个细胞膜片钳技术允许在电压钳条件下记录整个细胞的膜电流。然而，技术上的困难出现在具有广泛突的大细胞中，如培养的人类胎儿背根神经节（DRG）神经元。为了改善空间夹紧条件，将培养1-2周的人胎儿DRG神经元酶解分离并在新培养皿中复制，培养24 h后产生圆形或椭圆形细胞，无突起或短突起。电流钳记录显示，与对照非复制细胞相比，复制细胞的动作电位参数没有差异。被动特性分析表明，再生细胞的平均比膜电容减少40%，平均比膜电阻增加57%，这与细胞膜表面积的减少一致。全细胞电压钳研究表明空间钳有很大的改进，表明通过复制消除中性细胞可以在培养的神经元中实现更有效的电压钳条件。
The whole cell patch clamp technique allows recording of membrane currents in an entire cell under voltage clamp conditions. However, technical difficulties arise in large cells bearing extensive processes, such as human fetal dorsal root ganglion (DRG) neurons in culture. In order to improve space clamp conditions, human fetal DRG neurons cultured for 1-2 weeks were enzymatically detached and replated in new dishes, yielding round or oval cells with absent or short processes at 24 h in culture. Current clamp recordings demonstrated no difference in action potential parameters of the replated cells compared to control non-replated cells. Analysis of passive properties showed a reduction of 40% in mean specific membrane capacitance and a 57% increase in mean specific membrane resistance in the replated cells, consistent with the decrease of cell membrane surface area. Whole cell voltage clamp studies demonstrated great improvement of the space clamp, indicating that more efficient voltage clamp conditions can be achieved in neurons in culture by eliminating neutrites through replating.