title: Combined FISH and immunofluorescent staining methods to co-localize proteins and mRNA in neurons and brain tissue.
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.
authors.affiliation: Department of Microbiology, New York University School of Medicine, New York, NY, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Ma
authors.forename: Naoko
authors.initials: N
authors.lastname: Tanese
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology*; D005455:Fluorescent Antibody Technique / Q000379:methods*; D021621:Imaging, Three-Dimensional; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D051379:Mice; D018613:Microscopy, Confocal; D009474:Neurons / Q000378:metabolism*; D021381:Protein Transport; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D059629:Signal-To-Noise Ratio; D013194:Staining and Labeling / Q000379:methods*; D014162:Transfection
chemical_list: D011506:Proteins; D012333:RNA, Messenger