Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, 30 Convent Dr. 303A, Bethesda, MD 20892-4340, USA.
Traffic. 2012 May;13(5):627-34. doi: 10.1111/j.1600-0854.2012.01328.x. Epub 2012 Jan 31.
Regulated exocytosis is a fundamental process that every secretory cell uses to deliver molecules to the cell surface and the extracellular space by virtue of membranous carriers. This process has been extensively studied using various approaches such as biochemistry, electrophysiology and electron microscopy. However, recent developments in time-lapse light microscopy have made possible imaging individual exocytic events, hence, advancing our understanding of this process at a molecular level. In this review, we focus on intravital microscopy (IVM), a light microscopy-based approach that enables imaging subcellular structures in live animals, and discuss its recent application to study regulated exocytosis. IVM has revealed differences in regulation and modality of regulated exocytosis between in vitro and in vivo model systems, unraveled novel aspects of this process that can be appreciated only in in vivo settings and provided valuable and novel information on its molecular machinery. In conclusion, we make the case for IVM being a mature technique that can be used to investigate the molecular machinery of several intracellular events under physiological conditions.
受调控的胞吐作用是一种基本过程,每个分泌细胞都利用膜性载体将分子递送到细胞表面和细胞外空间。已经使用各种方法(如生物化学、电生理学和电子显微镜)对该过程进行了广泛研究。然而,延时荧光显微镜的最新发展使得对单个胞吐事件进行成像成为可能,从而使我们能够在分子水平上加深对该过程的理解。在这篇综述中,我们将重点介绍活体显微镜(IVM),这是一种基于荧光显微镜的方法,能够对活体动物中的亚细胞结构进行成像,并讨论其最近在研究受调控的胞吐作用中的应用。IVM 揭示了在体外和体内模型系统中受调控的胞吐作用的调节和模式的差异,揭示了该过程在体内环境中才能被认识到的新方面,并提供了关于其分子机制的有价值和新颖的信息。总之,我们认为 IVM 是一种成熟的技术,可以用于在生理条件下研究几种细胞内事件的分子机制。