Domire Jacqueline S, Mykytyn Kirk
Department of Pharmacology, Department of Internal Medicine Division of Human Genetics, College of Medicine, The Ohio State University, Columbus, Ohio 43210, USA.
Methods Cell Biol. 2009;91:111-21. doi: 10.1016/S0091-679X(08)91006-2. Epub 2009 Dec 1.
Primary cilia were first detected on neurons in the mammalian brain over 40 years ago using electron microscopy. However, this approach is very labor intensive and has inherent limitations that restrict its utility for studying neuronal cilia. While the study of cilia in other tissues was greatly facilitated by the identification of specific ciliary markers, historically there have been no markers for neuronal cilia. Fortunately, recent developments make the study of neuronal cilia more practical. First, specific proteins have been shown to selectively localize to neuronal cilia and can serve as markers by immunolabeling. Second, neurons have been shown to possess cilia in culture, which allows for the use of additional approaches, such as live-cell imaging of neuronal cilia. This chapter provides an overview of the current techniques for visualizing neuronal cilia in tissue as well as fixed and living cells. These approaches allow for the identification of additional neuronal ciliary proteins and provide a basis for future functional studies.
40多年前,人们首次使用电子显微镜在哺乳动物大脑的神经元上检测到初级纤毛。然而,这种方法非常耗费人力,并且存在固有的局限性,限制了其在研究神经元纤毛方面的效用。虽然通过鉴定特定的纤毛标记物极大地促进了对其他组织中纤毛的研究,但从历史上看,一直没有神经元纤毛的标记物。幸运的是,最近的进展使神经元纤毛的研究变得更加可行。首先,已证明特定蛋白质可选择性地定位于神经元纤毛,并可通过免疫标记用作标记物。其次,已证明神经元在培养物中具有纤毛,这使得可以使用其他方法,例如对神经元纤毛进行活细胞成像。本章概述了目前用于可视化组织以及固定和活细胞中神经元纤毛的技术。这些方法有助于鉴定更多的神经元纤毛蛋白,并为未来的功能研究提供基础。