Gladstone Institute of Virology and Immunology, San Francisco, CA 94158-2216, USA.
Methods. 2011 Jan;53(1):27-33. doi: 10.1016/j.ymeth.2010.04.002. Epub 2010 Apr 9.
A wide variety of cell types, including immune cells, have been observed to frequently interact via transient, long-distance membrane connections. However, considerable heterogeneity in their structure, mode of formation and functional properties has emerged, suggesting the existence of distinct subclasses. Open-ended tunneling nanotubes allow for the trafficking of cytoplasmic material, e.g. endocytic vesicles, or the transmission of calcium signals. Closed-ended membrane nanotubes do not seamlessly connect the cytoplasm between two interacting cells and a junction exists within the nanotube or where the nanotube meets a cell body. Recent live cell imaging suggested that membrane nanotubes between T cells could present a novel route for HIV-1 transmission. Here, we describe detailed protocols for observing membrane nanotubes and HIV-1 trafficking by live cell fluorescence microscopy.
多种细胞类型，包括免疫细胞，已经被观察到通过短暂的、长距离的膜连接频繁地相互作用。然而，它们的结构、形成方式和功能特性存在着相当大的异质性，这表明存在着不同的亚类。开放式的隧道纳米管允许细胞质物质（如内吞小泡）的运输，或钙信号的传递。封闭式的膜纳米管不能在两个相互作用的细胞之间无缝连接细胞质，并且在纳米管内或纳米管与细胞体相遇的地方存在连接。最近的活细胞成像表明，T 细胞之间的膜纳米管可能为 HIV-1 的传播提供了一种新途径。在这里，我们描述了通过活细胞荧光显微镜观察膜纳米管和 HIV-1 运输的详细方案。
