Vieillard Jennifer, Duteyrat Jean-Luc, Cortier Elisabeth, Durand Bénédicte
Centre de Génétique et de Physiologie Moléculaire et Cellulaire, Université Claude Bernard Lyon-1, Lyon, Villeurbanne, France.
Methods Cell Biol. 2015;127:279-302. doi: 10.1016/bs.mcb.2014.12.009. Epub 2015 Feb 14.
Drosophila melanogaster is a powerful genetic model organism to understand the function of proteins in specific cellular processes. Cilia have been extensively studied in Drosophila playing various sensory functions that are essential for fly survival. Indeed, flies defective in cilia formation cannot walk, fly, or feed properly. Drosophila harbors different types of cilia that can be motile or immotile or that can show compartimentalized (intraflagellar transport (IFT)-dependent) or cytoplasmic (IFT-independent) mode of assembly. Therefore, Drosophila represents an advantageous model organism to study the function of novel ciliary candidates and to address specific questions such as their requirement for IFT-dependent processes versus other aspects of cilia-associated functions. This chapter describes protocols to visualize cilia by direct or indirect fluorescent labeling and protocols to analyze ciliary ultrastructure by electron microscopy.
黑腹果蝇是一种强大的遗传模式生物，有助于理解蛋白质在特定细胞过程中的功能。在果蝇中，纤毛已得到广泛研究，发挥着各种对果蝇生存至关重要的感官功能。事实上，纤毛形成有缺陷的果蝇无法正常行走、飞行或进食。果蝇拥有不同类型的纤毛，这些纤毛可以是能动的或不能动的，或者可以呈现出分区化（依赖鞭毛内运输（IFT））或细胞质（不依赖IFT）的组装模式。因此，果蝇是研究新型纤毛候选物功能以及解决特定问题（如它们对依赖IFT的过程与纤毛相关功能的其他方面的需求）的有利模式生物。本章描述了通过直接或间接荧光标记可视化纤毛的方案以及通过电子显微镜分析纤毛超微结构的方案。
