Gene expression analysis of CCN proteins: Whole-mount in situ hybridization of Ccn2 in developing calcified tissues

Tomoichiro Yamaai; Masaharu Takigawa

doi:10.1007/978-1-4939-6430-7_2

Gene expression analysis of CCN proteins: Whole-mount in situ hybridization of Ccn2 in developing calcified tissues

Tomoichiro Yamaai, Masaharu Takigawa

Dental School

Research output: Chapter in Book/Report/Conference proceeding › Chapter

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.

Original language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	11-19
Number of pages	9
DOIs	https://doi.org/10.1007/978-1-4939-6430-7_2
Publication status	Published - 2017

Publication series

Name	Methods in Molecular Biology
Volume	1489
ISSN (Print)	1064-3745

Keywords

Ccn2 gene expression
Dig-labeled riboprobe
Mouse embryo
Whole-mount in situ hybridization

ASJC Scopus subject areas

Molecular Biology
Genetics

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10.1007/978-1-4939-6430-7_2

Cite this

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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.",

keywords = "Ccn2 gene expression, Dig-labeled riboprobe, Mouse embryo, Whole-mount in situ hybridization",

author = "Tomoichiro Yamaai and Masaharu Takigawa",

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language = "English",

series = "Methods in Molecular Biology",

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AU - Yamaai, Tomoichiro

AU - Takigawa, Masaharu

N1 - Publisher Copyright: © Springer Science+Business Media New York 2017.

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

KW - Ccn2 gene expression

KW - Dig-labeled riboprobe

KW - Mouse embryo

KW - Whole-mount in situ hybridization

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Gene expression analysis of CCN proteins: Whole-mount in situ hybridization of Ccn2 in developing calcified tissues

Abstract

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Keywords

ASJC Scopus subject areas

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