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杀虫剂溴氰菊酯对昆虫神经元作用的膜片钳分析

Patch-clamp analysis of the effects of the insecticide deltamethrin on insect neurones.

作者信息

Amar M, Pichon Y, Inoue I

机构信息

Département de Biophysique, Laboratoire de Neurobiologie Cellulaire et Moléculaire du CNRS, Gif sur Yvette Cedex, France.

出版信息

J Exp Biol. 1992 Feb;163:65-84. doi: 10.1242/jeb.163.1.65.

DOI:10.1242/jeb.163.1.65
PMID:1372926
Abstract
  1. The mode of action of the pyrethroid insecticide deltamethrin on inexcitable embryonic cultured cockroach neurones has been investigated using the patch-clamp technique. 2. Whole-cell recordings of the current induced by step depolarizations of the cell membrane showed that concentrations of deltamethrin ranging from 10(-8) to 5 x 10(-6) mol l-1 induced a small tetrodotoxin (TTX)-sensitive inward current that peaked at around +10 mV and reversed at around +60 mV. The activation and inactivation kinetics of this current were much slower than those of the axonal sodium current in this same species and were relatively insensitive to membrane potential. Steady-state inactivation was almost absent. 3. Single-channel activity associated with the action of the insecticide was analyzed using the cell-attached configuration. Three distinct patterns of activity were found: (1) discrete single-channel events of relatively short duration, (2) long events of comparatively small amplitude and (3) complex bursts made up of a succession of openings and closings to several levels. These three patterns were analyzed quantitatively using specially designed programs. 4. The first pattern of activity could be seen in most patches. It consisted of short (1-10 ms) rectangular events of comparatively small amplitude (1.5 pA at rest) and very low open time probability (around 0.001). The current-voltage relationship of these small events was linear over the voltage range studied and the (extrapolated) reversal potential approximated ENa. 5. The second pattern of activity was observed less frequently. The channels could stay open for very long periods (up to several seconds) and occasionally flickered between two or more levels. 6. The third pattern of activity was observed in many patches. During the burst, which could last from a few milliseconds to a few hundred milliseconds, the single-channel current jumped almost continuously between several levels (up to 7 or 8).
摘要
  1. 运用膜片钳技术研究了拟除虫菊酯类杀虫剂溴氰菊酯对不可兴奋的胚胎期培养蟑螂神经元的作用方式。2. 对细胞膜进行阶跃去极化所诱导电流的全细胞记录显示,浓度范围为10⁻⁸至5×10⁻⁶ mol l⁻¹的溴氰菊酯可诱导出一种小的对河豚毒素(TTX)敏感的内向电流,该电流在约 +10 mV 时达到峰值,并在约 +60 mV 时反转。此电流的激活和失活动力学比同一物种轴突钠电流的动力学要慢得多,并且对膜电位相对不敏感。几乎不存在稳态失活。3. 使用细胞贴附式记录构型分析了与杀虫剂作用相关的单通道活性。发现了三种不同的活性模式:(1)持续时间相对较短的离散单通道事件,(2)幅度相对较小的长事件,以及(3)由一系列在几个水平上的开放和关闭组成的复杂爆发。使用专门设计的程序对这三种模式进行了定量分析。4. 大多数膜片都能看到第一种活性模式。它由短(1 - 10毫秒)的矩形事件组成,幅度相对较小(静息时为1.5 pA),开放时间概率非常低(约0.001)。在研究的电压范围内,这些小事件的电流 - 电压关系呈线性,(外推的)反转电位接近ENa。5. 第二种活性模式较少观察到。通道可以长时间保持开放(长达几秒),偶尔在两个或更多水平之间闪烁。6. 在许多膜片中观察到了第三种活性模式。在持续时间从几毫秒到几百毫秒的爆发期间,单通道电流几乎连续地在几个水平(多达7或8个)之间跳跃。

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