Establishment Of Zebrafish Transgenic Strains And Preliminary Studies On Their Functions

The process of heart development is complex. Some signaling pathways, transcription factors and adhesion molecules regulate the proliferation, differentiation, the diversity and the morphogenesis of heart cells. We build some transgenic zebrafish models to research the biological functions of genes which highly express in heart tissues.It can lay a theoretical foundation for understanding the mechanism of heart development and the development of effective treatments.Report gene GFP and RFP is usually used to live report, track the protein expression of time and space. Therefore, we use them to study the genes associated with heart development. To12is a transposon system which was found in medaka. Constructing transgenic zebrafish models by using To12transposon system can significantly improve the efficiency of transgene. Consequently, To12transposon system provides a great convenience for studying the function of candidate genes in the zebrafish heart development.A gene can specific express in the heart and muscles, our laboratory screen and identify it HDGC3, which is a heart development candidate gene. Preliminary researches indicate that the HDGC3is high level expression in zebrafish heart and muscle tissue. In order to further study the function of HDGC3in the process of heart development, we constructed several cardiovascular transgenic zebrafish models basing on To12transgenic technology as well as the advantages of zebrafish. We have been construct two expression vectors:pTol2(cmlc2:HDGC3-EGFP) and pTol2(Titin:HDGC3-EGFP) by using cmlc2and Titin promoter. And then, we respectively mix two expression vectors with To12transposases according to certain proportion, then inject them to one-cell stage Zebrafish embryos. A few days afterwards, we screen and identify these embryos which have green fluorescence expression under the fluorescence microscope. So we have established two stable genetic zebrafish models:Tg (cmlc2:HDGC3-EGFP) and Tg (Titin:HDGC3-EGFP). They are used to preliminary analysis the influence on heart development and function in the HDGC3overexpression transgenic zebrafish models; At the same time, we cloned HDGC3gene promoter sequences, and Tg (HDGC3:EGFP) was established based on the promoter sequences. We examine the green fluorescence expression in the zebrafish embryo. The analysis results show that the HDGC3specific express in the early development of the heart and muscle. Tg (HDGC3: EGFP) was used to tracked the expression of HDGC3during the development of zebrafish. The transgenic zebrafish model is great significance for studying the function of HDGC3in heart development.