title: Gene Expression Analysis of CCN Proteins: Whole-Mount In Situ Hybridization of Ccn2 in Developing Calcified Tissues.
abstract: A procedure for whole-mount in situ hybridization developed for detecting gene expression of Ccn2 in developing calcified tissues of mouse embryos is presented. In this method, embryos are hybridized with Dig-labeled riboprobes, and the riboprobes are detected by use of the alkaline-phosphatase reaction in the presence of a 4-nitro-blue tetrazolium chloride and 5-bromo-4-chloro-3-indolyl-phosphate (NBT + BCIP) mixture. Obvious detection of positive signals for Ccn2 in the cartilage of developing phalanges indicates that this method can be applied to gene expression analysis of other Ccn genes in developing calcified tissues.
authors.affiliation: Department of Oral Function and Anatomy, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8558, Japan.
authors.forename: Tomoichiro
authors.initials: T
authors.lastname: Yamaai
authors.affiliation: Advanced Research Center for Oraland Craniofacial Sciences, Okayama University Dental School/Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1, Shikata-cho, Kita-ku, Okayama, 700-8525, Japan. takigawa@md.okayama-u.ac.jp.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Takigawa
mesh_terms: D000818:Animals; D055511:CCN Intercellular Signaling Proteins / Q000235:genetics* / Q000378:metabolism; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation; D017403:In Situ Hybridization / Q000379:methods; D051379:Mice; D013194:Staining and Labeling
chemical_list: D055511:CCN Intercellular Signaling Proteins
keywords: Ccn2 gene expression; Dig-labeled riboprobe; Mouse embryo; Whole-mount in situ hybridization