视紫红质通道蛋白2介导果蝇神经肌肉接头处突触电位的刺激。
Channelrhodopsin2 mediated stimulation of synaptic potentials at Drosophila neuromuscular junctions.
作者信息
Hornstein Nicholas J, Pulver Stefan R, Griffith Leslie C
机构信息
Department of Biology, Brandeis University, USA.
出版信息
J Vis Exp. 2009 Mar 16(25):1133. doi: 10.3791/1133.
Abstract
The Drosophila larval neuromuscular preparation has proven to be a useful tool for studying synaptic physiology. Currently, the only means available to evoke excitatory junctional potentials (EJPs) in this preparation involves the use of suction electrodes. In both research and teaching labs, students often have difficulty maneuvering and manipulating this type of stimulating electrode. In the present work, we show how to remotely stimulate synaptic potentials at the larval NMJ without the use of suction electrodes. By expressing channelrhodopsin2 (ChR2) in Drosophila motor neurons using the GAL4-UAS system, and making minor changes to a basic electrophysiology rig, we were able to reliably evoke EJPs with pulses of blue light. This technique could be of particular use in neurophysiology teaching labs where student rig practice time and resources are limited.
摘要
果蝇幼虫神经肌肉标本已被证明是研究突触生理学的有用工具。目前，在此标本中诱发兴奋性接头电位（EJP）的唯一可用方法是使用吸力电极。在研究实验室和教学实验室中，学生们经常在操作和操控这种刺激电极时遇到困难。在本研究中，我们展示了如何在不使用吸力电极的情况下远程刺激幼虫神经肌肉接头处的突触电位。通过使用GAL4-UAS系统在果蝇运动神经元中表达通道视紫红质2（ChR2），并对基本的电生理装置进行微小改动，我们能够用蓝光脉冲可靠地诱发EJP。这项技术在学生实验练习时间和资源有限的神经生理学教学实验室中可能会特别有用。
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本文引用的文献
1
Modification of a hydrophobic layer by a point mutation in syntaxin 1A regulates the rate of synaptic vesicle fusion.Syntaxin 1A 中的点突变对疏水层的修饰调节了突触小泡融合的速率。
PLoS Biol. 2007 Apr;5(4):e72. doi: 10.1371/journal.pbio.0050072.
2
Synaptic development: insights from Drosophila.突触发育：来自果蝇的见解
Curr Opin Neurobiol. 2007 Feb;17(1):35-42. doi: 10.1016/j.conb.2007.01.001. Epub 2007 Jan 16.
3
Light-induced activation of distinct modulatory neurons triggers appetitive or aversive learning in Drosophila larvae.光诱导不同调节神经元的激活触发果蝇幼虫的食欲或厌恶学习。
Curr Biol. 2006 Sep 5;16(17):1741-7. doi: 10.1016/j.cub.2006.07.023.
4
Light activation of channelrhodopsin-2 in excitable cells of Caenorhabditis elegans triggers rapid behavioral responses.秀丽隐杆线虫可兴奋细胞中视紫红质通道蛋白-2的光激活引发快速行为反应。
Curr Biol. 2005 Dec 20;15(24):2279-84. doi: 10.1016/j.cub.2005.11.032.
5
Channelrhodopsin-2, a directly light-gated cation-selective membrane channel.通道视紫红质-2，一种直接受光门控的阳离子选择性膜通道。
Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):13940-5. doi: 10.1073/pnas.1936192100. Epub 2003 Nov 13.
6
The drosophila neuromuscular junction: a model system for studying synaptic development and function.果蝇神经肌肉接头：用于研究突触发育和功能的模型系统。
Annu Rev Neurosci. 1996;19:545-75. doi: 10.1146/annurev.ne.19.030196.002553.
7
Targeted neuronal cell ablation in the Drosophila embryo: pathfinding by follower growth cones in the absence of pioneers.
Neuron. 1995 Apr;14(4):707-15. doi: 10.1016/0896-6273(95)90215-5.
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