How to Study Gene Expression and Gain of Function of Hoxb1 in Mouse Heart Development

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de Bono, Christopher | Lescroart, Fabienne | Zaffran, Stéphane

Edité par CCSD ; Springer US -

International audience. Anterior Hox genes are required for genetic identity and anterior posterior patterning of the second heartfield (SHF), which contributes to the formation of the embryonic heart in vertebrates. Defective contribu-tion of SHF cells to the arterial or venous pole of the heart is often associated with severe congenital heartdefects. The mouse Cre-lox system allows the activation of expression of any gene of interest in restrictedtissues. We developed a gain of function approach that relies on the use of a CAG transgene to ectopicallyactivate Hoxb1 expression in SHF cardiac progenitor cells through specific Cre activation. Therefore, wegenerated a floxed transgenic mouse line, CAG-Hoxb1-eGFP, which upon recombination by Cre recombi-nase conditionally induces robust Hoxb1 and eGFP expression. When induced within the anterior SHFlineage, we detected heart defects in mouse embryos such as right ventricular hypoplasia. Here, we describethe strategy for generating and genetically crossing this transgenic mouse line. We also provide detailedprotocols for whole-mount embryo and paraffin section in situ RNAscope hybridization and X-gal stainingallowing investigation of SHF contribution during heart development.

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