Abstract
Disconnected (disco)-interacting protein 2 homolog A is a member of the DIP2 protein family encoded by Dip2a gene. Dip2a expression pattern has never been systematically studied. Functions of Dip2a in embryonic development and adult are not known. To investigate Dip2a gene expression and function in embryo and adult, a Dip2a-LacZ mouse model was generated by insertion of β-Gal cDNA after Dip2a promoter using CRISPR/Cas9 technology. Dip2a-LacZ mouse was designed to be a lacZ reporter mouse as well as a Dip2a knockout mouse. Heterozygous mice were used to study endogenous Dip2a expression and homozygotes to study DIP2A-associated structure and function. LacZ staining indicated that Dip2a is broadly expressed in neuronal, reproductive and vascular tissues, as well as in heart, kidney, liver and lung. Results demonstrate that Dip2a is expressed in ectoderm-derived tissues in developing embryos. Adult tissues showed rich staining in neurons, mesenchymal, endothelial, smooth muscle cells and cardiomyocytes by cell types. The expression pattern highly overlaps with FSTL1 and supports previous report that DIP2A to be potential receptor of FSTL1 and its protective roles of cardiomyocytes. Broad and intense embryonic and adult expression of Dip2a has implied their multiple structural and physiological roles.
Highlights
DIP2A is a member of Disconnected-interacting protein 2 (DIP2) family with other two isoforms, DIP2B and DIP2C
Dip2a-LacZ-neo mouse was crossed with transgenic mice that express Flipase to excise Neo cassette, resulting to Dip2aLacZ/+ mice [7]
Genetic background of Dip2aLacZ/+ mice was switched to C57BL/6J by continued crossing heterozygous Dip2aLacZ/+ mice with C57BL/6J mice
Summary
DIP2A is a member of Disconnected (disco)-interacting protein 2 (DIP2) family with other two isoforms, DIP2B and DIP2C. Bioinformatic analysis using Predict Protein and Homolo Gene suggested that DIP2A is a type I receptor molecule with DMAP, CaiC and AMP-binding domains [1]. Mukhopadhyay reported that DIP2 homologs are evolutionarily conserved in organisms from C. elegans to humans. DIP2A proteins may exert their signaling roles as receptors in prokaryotes and eukaryotes and may provide positional cues for axon path finding and patterning [2]. Expression pattern and physiological function of DIP2A are currently.
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