Cardiomyocytes, the heart’s muscle cells, are a vital part of heart function as they enable the heart’s ability to pump. During heart development these cardiomyocytes are surrounded by an extracellular matrix (ECM). Integrins are transmembrane receptors that bind to the ECM. They activate signal transduction pathways, which play an important role in various cellular processes. Zebrafish double mutants in the Fibronectin (Fn1) receptors Integrin alpha5 (Itga5) and Integrin alpha4 (Itga4) display defects in various heart tissues such as cardia bifida and failure of endocardial differentiation. To investigate how Integrins specifically function in the cardiomyocytes, we generated a transgenic plasmid that expresses the Fn1 receptor Itga5 specifically in the myocardium of the heart. We utilized Gateway cloning technology to combine the gene promoter p5E-cmcl2, the Integrin pME-itga5, and the fluorescent tag p3E-H2A-mCherry to create our transgenic plasmid. After analyzing our restriction digests we confirmed the Gateway assembly was successful and obtained the plasmid we were seeking. Looking ahead we will inject the obtained plasmid into the Zebrafish embryo to obtain a stable transgenic line.
Author(s): Skylie Antle, Lily Lerman, Jennifer A. Schumacher
Advisor(s): Jennifer A. Schumacher, Biology
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