title: Non-isotopic RNA In Situ Hybridization on Embryonic Sections.
abstract: This unit describes methods for non-isotopic RNA in situ hybridization on embryonic mouse sections. These methods can be used to follow the spatiotemporal dynamics of gene expression in an embryonic tissue of interest. They involve the use of labeled (e.g., digoxygenin, FITC) antisense riboprobes that hybridize to a specific mRNA in the target tissue. The probes are detected using an alkaline phosphatase-conjugated antibody recognizing the label and a chromogenic substrate. This method can be used to: (1) assess the expression of a single gene within a tissue, (2) compare the expression profiles of two genes within a tissue, or (3) compare the distribution of a transcript and protein within a tissue. While this approach is not quantitative, it provides a qualitative assessment of the precise cell types where a gene is expressed, which is not easily achievable with other more quantitative methods such as quantitative PCR.
authors.affiliation: Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children's Hospital Research Institute, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Yacine
authors.initials: Y
authors.lastname: Touahri
authors.affiliation: Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children's Hospital Research Institute, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Lata
authors.initials: L
authors.lastname: Adnani
authors.affiliation: Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children's Hospital Research Institute, University of Calgary, Calgary, Alberta, Canada.|Current address: Cellular Neurobiology Research Unit, Institut de Recherches Cliniques de Montréal, Montréal, Quebec, Canada.
authors.forename: Pierre
authors.initials: P
authors.lastname: Mattar
authors.affiliation: Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children's Hospital Research Institute, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Markham
authors.affiliation: Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children's Hospital Research Institute, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Natalia
authors.initials: N
authors.lastname: Klenin
authors.affiliation: Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children's Hospital Research Institute, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Carol
authors.initials: C
authors.lastname: Schuurmans
mesh_terms: D000818:Animals; D004622:Embryo, Mammalian / Q000737:chemistry* / Q000378:metabolism*; D005821:Genetic Techniques; D017403:In Situ Hybridization; D051379:Mice; D015347:RNA Probes / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D015347:RNA Probes
keywords: embryonic mouse; gene expression; in situ hybridization; mRNA detection; spatiotemporal dynamics