TY - JOUR A1 - Blanco Fernández de Valderrama, María José AU - Rodríguez Learte, Ana Isabel AU - Marchena, Miguel Ángel AU - Muñoz Sáez, Emma AU - Cid Torres, María Antonia AU - Rodríguez Martín, Iván AU - Sánchez-Camacho Blázquez, Cristina T1 - Tracing Gene Expression Through Detection of β-galactosidase Activity in Whole Mouse Embryos Y1 - 2018 SN - 1940-087X UR - http://hdl.handle.net/11268/7957 AB - The Escherichia coli LacZ gene, encoding β-galactosidase, is largely used as a reporter for gene expression and as a tracer in cell lineage studies. The classical histochemical reaction is based on the hydrolysis of the substrate X-gal in combination with ferric and ferrous ions, which produces an insoluble blue precipitate that is easy to visualize. Therefore, β-galactosidase activity serves as a marker for the expression pattern of the gene of interest as the development proceeds. Here we describe the standard protocol for the detection of β-galactosidase activity in early whole mouse embryos and the subsequent method for paraffin sectioning and counterstaining. Additionally, a procedure for clarifying whole embryos is provided to better visualize X-gal staining in deeper regions of the embryo. Consistent results are obtained by performing this procedure, although optimization of reaction conditions is needed to minimize background activity. Limitations in the assay should be also considered, particularly regarding the size of the embryo in whole mount staining. Our protocol provides a sensitive and a reliable method for β- galactosidase detection during the mouse development that can be further applied to the cryostat sections as well as whole organs. Thus, the dynamic gene expression patterns throughout development can be easily analyzed by using this protocol in whole embryos, but also detailed expression at the cellular level can be assessed after paraffin sectioning. KW - Genética KW - Cerebro KW - Genética KW - Cerebro LA - eng ER -