研究乙醇对成年哺乳动物神经元离子电流的急性和慢性影响的电压钳模型。

S R Ikeda, G G Schofield, F F Weight
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引用次数: 0

摘要

本研究的目的是开发和表征一个模型系统,在该模型系统中,乙醇对成年哺乳动物神经元中电压和激动剂门控离子电流的影响可以使用电压箝位技术进行研究。我们发现,从成年大鼠和豚鼠的外周神经系统(结节和颈上神经节)和中枢神经系统(海马锥体层)中酶解分离的神经元比传统的神经元制备(例如,完整的神经节或脑切片)有几个优点。首先,分离的神经元与膜片钳技术相结合,可以高保真地记录宏观和单通道电流。其次,电流和电压钳记录显示,分离神经元的主动和被动膜特性、化学敏感性和离子电流与传统制备中描述的相似。最后,我们开发了一种细胞内灌注系统,可以方便地控制细胞内环境。这一技术对离子电流下细胞内第二信使的研究是有用的。我们的研究结果表明,分离的成年哺乳动物神经元非常适合研究乙醇对膜兴奋性的急性和慢性影响。
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Voltage-clamp models for the study of acute and chronic effects of ethanol on ionic currents in adult mammalian neurons.

The aim of this study was to develop and characterize a model system in which the effects of ethanol on voltage- and agonist-gated ionic currents in adult mammalian neurons could be studied using voltage-clamp techniques. We have found that neurons enzymatically isolated from the peripheral (nodose and superior cervical ganglia) and central nervous system (pyramidal layer of the hippocampus) of the adult rat and guinea pig provide several advantages over conventional neuronal preparations (e.g., intact ganglia or brain slice). First, the isolated neurons, in conjunction with the patch clamp technique, allow high fidelity recordings of both macroscopic and single channel currents. Secondly, current- and voltage-clamp recordings have revealed that active and passive membrane properties, chemosensitivity, and ionic currents in the isolated neurons resemble those described from conventional preparations. Finally, we have developed an intracellular perfusion system which allows the convenient control of the intracellular milieu. This technique should be useful for the study of intracellular second messengers on ionic currents. Our results demonstrate that isolated adult mammalian neurons are ideally suited for the study of both the acute and chronic effects of ethanol on membrane excitability.

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