Sweeney Sean T, Hidalgo Alicia, de Belle J Steven, Keshishian Haig
Cold Spring Harb Protoc. 2012 Feb 1;2012(2):239-41. doi: 10.1101/pdb.prot067405.
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. β-Galactosidase, the product of the E. coli lacZ gene, has been used extensively as a reporter in Drosophila research. Staining for β-galactosidase activity can be performed using the substrate X-gal (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside), which produces a blue precipitate visible by light microscopy. This detection method is highly sensitive and has the advantage that the results can be observed without the need for specialized microscopy equipment. This protocol describes general procedures for X-gal labeling of neural tissue from Drosophila.
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。胚胎后期的果蝇大脑尤其有用，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，可为我们理解发育神经生物学中涉及的遗传和分子机制做出重大贡献。β-半乳糖苷酶是大肠杆菌lacZ基因的产物，在果蝇研究中已被广泛用作报告基因。可以使用底物X-gal（5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷）对β-半乳糖苷酶活性进行染色，该底物会产生一种蓝色沉淀，在光学显微镜下可见。这种检测方法高度灵敏，优点是无需专门的显微镜设备即可观察结果。本方案描述了对果蝇神经组织进行X-gal标记的一般程序。
