Andlauer Till F M, Sigrist Stephan J
Cold Spring Harb Protoc. 2012 Apr 1;2012(4):481-9. doi: 10.1101/pdb.prot068593.
In the past decade, a significant number of proteins involved in the developmental assembly and maturation of synapses have been identified. However, detailed knowledge of the molecular processes underlying developmental synapse assembly is still sparse. We have developed an approach that makes extended in vivo imaging of selected proteins in live Drosophila larvae feasible at a single-synapse resolution. This protocol describes the repetitive, noninvasive imaging of the neuromuscular junction (NMJ) of a live, intact, anesthetized Drosophila larva over extended periods of time with confocal microscopy. Proteins of interest must be tagged with a fluorescent label and have to be expressed in transgenic fly strains. The method has proven highly useful for the study of synaptic assembly and the trafficking of proteins. These data contribute to our understanding of synaptic assembly under in vivo conditions.
在过去十年中，已经鉴定出大量参与突触发育组装和成熟的蛋白质。然而，关于发育性突触组装背后分子过程的详细知识仍然匮乏。我们开发了一种方法，使得在活的果蝇幼虫中以单突触分辨率对选定蛋白质进行长时间的体内成像成为可能。本方案描述了使用共聚焦显微镜对活的、完整的、麻醉的果蝇幼虫的神经肌肉接头（NMJ）进行长时间的重复、非侵入性成像。感兴趣的蛋白质必须用荧光标签标记，并在转基因果蝇品系中表达。该方法已被证明对研究突触组装和蛋白质运输非常有用。这些数据有助于我们理解体内条件下的突触组装。
