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电鳗电细胞的膜片钳记录。钠电流及钠钾通透率的变异性。

Patch recordings from the electrocytes of Electrophorus electricus. Na currents and PNa/PK variability.

作者信息

Shenkel S, Sigworth F J

机构信息

Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06510.

出版信息

J Gen Physiol. 1991 May;97(5):1013-41. doi: 10.1085/jgp.97.5.1013.

Abstract

Sodium currents were recorded in cell-attached and inside-out patches from the innervated membrane of Electrophorus electrocytes. Electrocytes from Sachs and main electric organs were prepared as described by Pasquale et al. (1986. J. Membr. Biol. 93:195.). Maximal currents in the Sachs organ, measured with 1-2 microns diameter patch pipettes and at room temperature, were in the range of 20 to 300 pA (27 patches) and were obtained near +10 mV. This range of current corresponds to approximately 70 to 1,300 channels in a patch. Maximal current in main organ cells also occurred near +10 mV and were in the range of 100 to 400 pA. Delayed K current was observed in a few patches. The inactivation phase of the currents during maintained depolarizations appears to be a single-exponential relaxation. The time constant decreases from 1 ms near -55 mV to a minimum of 0.3 ms near 0 mV, and then gradually increases with stronger depolarization. The mean currents are half inactivated near -90 mV with an apparent voltage dependence of e-fold per 6 mV. No apparent differences were observed in the decay time course or steady-state inactivation of the currents in the same patch before and after excision. From ensemble fluctuation analysis the peak open probability was found to be approximately 0.5 at +25 mV and increased only gradually with larger depolarizations. The single channel conductances were approximately 20 pS with 200 mM Na outside and 200 mM K inside, and 40 pS in 400 mM solutions. Reversal potentials in the 200 Na parallel 200 K solutions ranged from +51 to +94 mV in multichannel patches, corresponding to selectivity ratios PNa/PK from 8 to 43. Large differences in reversal potentials were seen even among patches from the same cell. Several controls rule out obvious sources of error in the reversal potential measurements. It is concluded that there is heterogeneity in the selectivity properties of the Na channels.

摘要

在电鳗电细胞的受神经支配膜的细胞贴附式和内面向外式膜片中记录钠电流。按照帕斯夸莱等人(1986年,《膜生物学杂志》93卷:195页)所述制备来自萨克斯氏器官和主要发电器官的电细胞。在室温下,用直径1 - 2微米的膜片吸管在萨克斯氏器官中测得的最大电流在20至300皮安范围内(27个膜片),且在约 +10毫伏时获得。该电流范围对应于一个膜片中约70至1300个通道。主要器官细胞中的最大电流也出现在约 +10毫伏附近,范围为100至400皮安。在少数膜片中观察到延迟钾电流。在持续去极化期间电流的失活阶段似乎是单指数弛豫。时间常数从 -55毫伏附近的1毫秒降至0毫伏附近的最小值0.3毫秒,然后随着更强的去极化逐渐增加。平均电流在 -90毫伏附近半失活,表观电压依赖性为每6毫伏一个e倍。在切除前后,同一膜片中电流的衰减时间进程或稳态失活未观察到明显差异。通过整体波动分析发现,在 +25毫伏时峰值开放概率约为0.5,并且随着更大的去极化仅逐渐增加。在外部为200毫摩尔钠且内部为200毫摩尔钾时,单通道电导约为20皮秒,在400毫摩尔溶液中为40皮秒。在多通道膜片中,200钠平行200钾溶液中的反转电位范围为 +51至 +94毫伏,对应于选择性比PNa/PK从8至43。即使在来自同一细胞的膜片之间,反转电位也存在很大差异。几个对照排除了反转电位测量中明显的误差来源。得出结论,钠通道的选择性特性存在异质性。

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