Sweeney Sean T, Hidalgo Alicia, de Belle J Steven, Keshishian Haig
Cold Spring Harb Protoc. 2012 Feb 1;2012(2):235-8. doi: 10.1101/pdb.prot067397.
The Drosophila nervous system provides a valuable model for studying various aspects of brain development and function. The postembryonic Drosophila brain is especially useful, because specific neuron types derive from specific progenitors at specific times. Elucidating the means by which diverse neuron types derive from a limited number of progenitors can contribute significantly to our understanding of the genetic and molecular mechanisms involved in developmental neurobiology. Antibody-labeling techniques are particularly useful for examining the Drosophila brain. These methods generally use primary antibodies specific to a protein or a structure of interest and a fluorescently labeled or enzyme-coupled secondary antibody to detect the primary antibodies. Immunofluorescence methods allow for simultaneous probing for multiple antigens using different fluorophores, as well as high-resolution confocal examination of deep structures. This protocol describes general procedures for antibody labeling of neural tissue from Drosophila, as well as visualization techniques for fluorescent and enzyme-linked probes.
果蝇神经系统为研究大脑发育和功能的各个方面提供了一个有价值的模型。果蝇胚胎后期的大脑尤其有用，因为特定的神经元类型在特定时间源自特定的祖细胞。阐明多种神经元类型如何从有限数量的祖细胞衍生而来，能够极大地促进我们对发育神经生物学中涉及的遗传和分子机制的理解。抗体标记技术对于研究果蝇大脑特别有用。这些方法通常使用针对感兴趣的蛋白质或结构的一抗，以及荧光标记或酶联二抗来检测一抗。免疫荧光方法允许使用不同的荧光团同时探测多种抗原，以及对深层结构进行高分辨率共聚焦检查。本方案描述了果蝇神经组织抗体标记的一般程序，以及荧光和酶联探针的可视化技术。
