1. Expression analysis of murine genes using in situ hybridization with radioactive and nonradioactively labeled RNA probes
- Author
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Anne, Chotteau-Lelièvre, Pascal, Dollé, Françoise, Gofflot, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Louis Pasteur - Strasbourg I, Toussaint, Jean-Luc, Institut de génétique et biologie moléculaire et cellulaire ( IGBMC ), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), and Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
MESH: Gene Expression ,MESH: Paraffin Embedding ,Gene Expression ,MESH : RNA Probes ,[ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,MESH: Histological Techniques ,In Vitro Techniques ,[SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Sensitivity and Specificity ,MESH : Frozen Sections ,Mice ,MESH: Pregnancy ,MESH: In Situ Hybridization ,Pregnancy ,MESH : Mice ,Animals ,Frozen Sections ,MESH : Female ,MESH: Animals ,MESH: Mice ,In Situ Hybridization ,Paraffin Embedding ,MESH : Embryo ,MESH: Frozen Sections ,MESH : Paraffin Embedding ,Histological Techniques ,MESH: Digoxigenin ,MESH: Embryo ,MESH : Digoxigenin ,MESH : In Situ Hybridization ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,RNA Probes ,Embryo, Mammalian ,MESH: Sensitivity and Specificity ,MESH : Gene Expression ,MESH : Pregnancy ,Female ,MESH : Histological Techniques ,MESH: RNA Probes ,MESH : Animals ,MESH : Sensitivity and Specificity ,Digoxigenin ,MESH: Female - Abstract
International audience; The term in situ hybridization (ISH) refers to all methods allowing the detection of specific DNA (gene loci) or RNA (gene expression products) sequences, using molecular hybridization (base pairing) of labeled nucleic acid probes to target molecules within "intact" cell populations in tissue sections or whole organisms, cultured cells, or chromosomal spreads. For more than two decades, ISH has been one of the main approaches used to characterize gene expression patterns in all laboratory animal models, especially in the context of embryonic development, as well as in human tissue or cell samples for both research and diagnostic purposes. Here, we describe several ISH protocols applied to the analysis of mouse embryos and tissues; this organism has become a reference for mammalian experimental genetics. These protocols use in vitro transcribed RNAs as probes for detection. Radiolabeled probes (using 35S as a radioisotope) allow sensitive ISH on sections of paraffin-embedded material, whereas nonradioactively (digoxigenin) labeled probes can be used both for hybridization of whole embryos (whole-mount ISH) and frozen tissue sections.
- Published
- 2006