Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
Genes Brain Behav. 2011 Nov;10(8):883-900. doi: 10.1111/j.1601-183X.2011.00729.x. Epub 2011 Oct 19.
Genetic manipulation of individual neurons provides a powerful approach toward understanding their contribution to stereotypic behaviors. We describe and evaluate a method for identifying candidate interneurons and associated neuropile compartments that mediate Drosophila larval locomotion. We created Drosophila larvae that express green fluorescent protein (GFP) and a shibire(ts1) (shi(ts1)) transgene (a temperature-sensitive neuronal silencer) in small numbers of randomly selected cholinergic neurons. These larvae were screened for aberrant behavior at an elevated temperature (31-32°C). Among larvae with abnormal locomotion or sensory-motor responses, some had very small numbers of GFP-labeled temperature-sensitive interneurons. Labeled ascending interneurons projecting from the abdominal ganglia to specific brain neuropile compartments emerged as candidates for mediation of larval locomotion. Random targeting of small sets of neurons for functional evaluation, together with anatomical mapping of their processes, provides a tool for identifying the regions of the central nervous system that are required for normal locomotion. We discuss the limitations and advantages of this approach to discovery of interneurons that regulate motor behavior.
个体神经元的遗传操作提供了一种强大的方法，可以深入了解它们对刻板行为的贡献。我们描述并评估了一种识别候选中间神经元及其相关神经丛室的方法，这些神经元和神经丛室介导果蝇幼虫的运动。我们创建了在少数随机选择的胆碱能神经元中表达绿色荧光蛋白（GFP）和 shibire(ts1)（shi(ts1)）转基因（一种温度敏感的神经元沉默物）的果蝇幼虫。这些幼虫在较高温度（31-32°C）下进行行为异常筛选。在运动或感觉运动反应异常的幼虫中，有些幼虫的 GFP 标记的温度敏感中间神经元数量非常少。从腹部神经节投射到特定脑神经丛室的标记的上升中间神经元成为幼虫运动调节的候选者。对小群神经元进行功能评估的随机靶向，以及它们过程的解剖映射，为识别中枢神经系统中调节运动行为所需的区域提供了一种工具。我们讨论了这种方法发现调节运动行为的中间神经元的局限性和优势。
