1] Centre for Research in Neuroscience, Research Institute of the McGill University Health Centre, Montréal, Québec, Canada. [2].
Nat Protoc. 2014 Jan;9(1):1-10. doi: 10.1038/nprot.2013.157. Epub 2013 Dec 5.
We describe a protocol to image the complex axonal branching structure of identified mechanosensory neurons in Drosophila, combined with a behavioral assay to evaluate the functional output of the neuron. The stimulation of identified mechanosensory neurons in live animals produces a stereotyped grooming reflex. The mechanosensory axonal arbor within the CNS is subsequently labeled with a lipophilic fluorescent dye and imaged by fluorescence microscopy. The behavioral output can therefore be correlated to the axonal morphology of the stimulated neuron in the same animal. Combining this protocol with genetic analysis provides a powerful tool for identifying the roles of molecules involved in different aspects of hard-wired neural circuit formation underlying an innate behavior. From behavioral analysis to axonal imaging, the protocol takes 4 d.
我们描述了一种方案，用于对果蝇中已鉴定的机械感觉神经元的复杂轴突分支结构进行成像，同时结合行为测定来评估神经元的功能输出。在活体动物中刺激已鉴定的机械感觉神经元会产生刻板的梳理反射。随后，用亲脂性荧光染料对中枢神经系统内的机械感觉轴突树突进行标记，并通过荧光显微镜进行成像。因此，可以将行为输出与同一动物中受刺激神经元的轴突形态相关联。将该方案与遗传分析相结合，为鉴定参与先天行为的固有神经回路形成的不同方面的分子的作用提供了强有力的工具。从行为分析到轴突成像，整个方案需要 4 天时间。
