Olsen Douglas P, Keshishian Haig
Cold Spring Harb Protoc. 2012 Feb 1;2012(2):226-30. doi: 10.1101/pdb.prot067801.
The Drosophila neuromuscular junction (NMJ) ranks as one of the preeminent model systems for studying synaptic development, function, and plasticity. This protocol describes the use of the two-electrode voltage clamp (TEVC) to examine potassium (K(+)) currents mediated by voltage-gated ion channels, and gives several genetic and pharmacological methods that are used to study the currents. Drosophila larval muscle fibers possess three major K(+) currents. One of these, a fast voltage-activating and inactivating I(A) current, is mediated by the Shaker channel. The Shaker channel is characterized by its sensitivity to the drug 4-aminopyridine (4-AP). Two useful transgenic tools for altering membrane excitability have been developed by making specific modifications of the Shaker channel; their use is described here.
果蝇神经肌肉接头（NMJ）是研究突触发育、功能和可塑性的卓越模型系统之一。本方案描述了使用双电极电压钳（TEVC）来检测电压门控离子通道介导的钾（K(+)）电流，并给出了几种用于研究这些电流的遗传学和药理学方法。果蝇幼虫肌肉纤维具有三种主要的K(+)电流。其中一种是快速电压激活和失活的I(A)电流，由Shaker通道介导。Shaker通道的特点是对药物4-氨基吡啶（4-AP）敏感。通过对Shaker通道进行特定修饰，开发了两种用于改变膜兴奋性的有用转基因工具；此处描述了它们的使用方法。
