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形态学鉴定的皮肤传入背根神经节神经元以不同比例表达三种不同的钾电流。

Morphologically identified cutaneous afferent DRG neurons express three different potassium currents in varying proportions.

作者信息

Everill B, Rizzo M A, Kocsis J D

机构信息

Department of Neurology, Yale University School of Medicine, New Haven 06510, USA.

出版信息

J Neurophysiol. 1998 Apr;79(4):1814-24. doi: 10.1152/jn.1998.79.4.1814.

Abstract

Outward K+ currents were recorded using a whole cell patch-clamp configuration, from acutely dissociated adult rat cutaneous afferent dorsal root ganglion (DRG) neurons (L4 and L5) identified by retrograde labeling with Fluoro-gold. Recordings were obtained 16-24 h after dissociation from cells between 39 and 49 mm in diameter with minimal processes. These cells represent medium-sized DRG neurons relative to the entire population, but are large cutaneous afferent neurons giving rise to myelinated axons. Voltage-activated K+ currents were recorded routinely during 300-ms depolarizing test pulses increasing in 10-mV steps from -40 to +50 mV; the currents were preceded by a 500-ms conditioning prepulse of either -120 or -40 mV. Coexpression of at least three components of K+ current was revealed. Separation of these components was achieved on the basis of sensitivities to the K+ channel blockers, 4-aminopyridine (4-AP) and dendrotoxin (DTx), and by the current responses to variation in conditioning voltage. Changing extracellular K+ concentration from 3 to 40 mM resulted in a shift to the right of the I-V curve commensurate with K+ being the principal charge carrier. Presentation of 100 mM 4-AP revealed a rapidly activating K+ current sensitive to low concentrations of 4-AP. High concentrations of 4-AP (6 mM) extinguished all inactivating current, leaving almost pure sustained current (IK). On the basis of the relative distribution of K+ currents neurons could be separated into three distinct categories: fast inactivating current (IA), slow inactivating current (ID), and sustained current (IK); only IA and IK; and slow inactivating current and IK. However, IK was always the dominant outward current component. These results indicate that considerable variation in K+ currents is present not only in the entire population of DRG neurons, as previously reported, but even within a restricted size and functional group (large cutaneous afferent neurons).

摘要

采用全细胞膜片钳配置记录外向钾电流,记录对象为经荧光金逆行标记鉴定的成年大鼠急性解离的皮肤传入背根神经节(DRG)神经元(L4和L5)。在细胞解离后16 - 24小时进行记录,选取直径在39至49μm之间且突起最少的细胞。相对于整个神经元群体,这些细胞代表中等大小的DRG神经元,但属于发出有髓轴突的大型皮肤传入神经元。在300毫秒的去极化测试脉冲期间常规记录电压激活的钾电流,测试脉冲从 - 40 mV到 + 50 mV以10 mV步长增加;电流之前有一个500毫秒的 - 120 mV或 - 40 mV的预处理脉冲。结果显示钾电流至少有三种成分共表达。基于对钾通道阻滞剂4 - 氨基吡啶(4 - AP)和树突毒素(DTx)的敏感性以及对预处理电压变化的电流响应,实现了这些成分的分离。将细胞外钾浓度从3 mM变为40 mM导致I - V曲线向右移动,这与钾作为主要电荷载体相符。施加100 mM的4 - AP可揭示一种对低浓度4 - AP敏感的快速激活钾电流。高浓度的4 - AP(6 mM)可消除所有失活电流,几乎只剩下纯的持续电流(IK)。根据钾电流的相对分布,神经元可分为三个不同类别:快速失活电流(IA)、缓慢失活电流(ID)和持续电流(IK);只有IA和IK;以及缓慢失活电流和IK。然而,IK始终是主要的外向电流成分。这些结果表明,不仅如先前报道的那样,DRG神经元的整个群体中钾电流存在相当大的变化,而且在一个受限的大小和功能组(大型皮肤传入神经元)内也是如此。

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