联合荧光原位杂交（FISH）和免疫荧光染色方法，以在神经元和脑组织中对蛋白质和mRNA进行共定位。

Combined FISH and immunofluorescent staining methods to co-localize proteins and mRNA in neurons and brain tissue.

作者信息

Ma Bin, Tanese Naoko

机构信息

Department of Microbiology, New York University School of Medicine, New York, NY, USA.

出版信息

Methods Mol Biol. 2013;1010:123-38. doi: 10.1007/978-1-62703-411-1_9.

DOI:10.1007/978-1-62703-411-1_9

PMID:23754223

Abstract

Combining multicolor fluorescent in situ hybridization (FISH) and immunofluorescent staining (IFS) presents a powerful method for visualizing the spatial relationship between mRNA and proteins in different neural compartments. Although seemingly straightforward, the combination of IFS/FISH and quantitative co-localization analysis of mRNA and proteins can be difficult to perform successfully, often generating variable results. Here we describe a combined method of multicolor IFS and FISH in concert with two-dimensional (2D) and three-dimensional (3D) co-localization analysis for determining the expression of individual molecules in rat neurons and brain sections. Using this approach, we have analyzed interactions of the Huntington's disease protein huntingtin with select proteins and mRNA.

摘要

将多色荧光原位杂交（FISH）与免疫荧光染色（IFS）相结合，为可视化不同神经区室中mRNA与蛋白质之间的空间关系提供了一种强大的方法。尽管看似简单，但IFS/FISH的结合以及mRNA与蛋白质的定量共定位分析往往难以成功实施，常常会产生不一致的结果。在此，我们描述了一种多色IFS与FISH相结合的方法，并结合二维（2D）和三维（3D）共定位分析，以确定大鼠神经元和脑切片中单个分子的表达情况。利用这种方法，我们分析了亨廷顿舞蹈病蛋白亨廷顿与特定蛋白质和mRNA之间的相互作用。