Mechanism Of Pygo1 And Nulp1 On Heart Development Of Zebrafish Analyzed By Whole Mount In Situ Hybridization

Heart development is an extremely complex process, subjecting to a series of precise control of the cardiac developmental genes. Mutations or abnormal expression of these genes will affect cardiomyocyte differentiation, diversity and pattern of myocardial cells, but their specific molecular mechanism remains to be elucidated. Classic Wnt signaling pathway (the Wnt-P-catenin signaling pathway) plays a very important role in embryonic development of organisms and the disease process.Pygol,a candidate gene for heart development which cloned by our laboratory, is a member of the Wnt signaling pathway and auxiliary active beta-catenin. In this article the mRNA level was studied to analyze the effect of pygol gene in heart development caused by expression change of specific gene markers of cardiac development, such as Amhc, Vmhc, Myl7, Nppa, Bmp4 and Hand2, detected by the method of the whole mount in situ hybridization with the zebrafish as a model in different developmental stages. The results showed that in the early and late stages of zebrafish development, pygol through regulating the expression level of Amhc, Vmhc Myll, Nppa, Bmp4, and Hand2 to influence the heart development of zebrafish.Nulpl, is also a candidate gene for heart development which cloned by our laboratory. In this paper, we used zebrafish as a model to study the role of nulpl gene in the different stages of zebrafish heart development. The morpholinos technology was uesd to knock down the expression level of nulp1 gene in two transgenic zebrafish, Tg(nppa::EGFP) and Tg (cmlc2::dsRed2), to analyze the role in vertebrate heart development. We found that when knocking down nulpl to study its function in late stages of zebrafish development, compared with the control, the heart looping is abnormal, and the size of ventricule become smaller, however, the atrium was bigger than control. Preliminary results indicate that nulpl can affect the heart development of zebrafish. The physiology of the zebrafish heart analysis also confirmed that nulpl can affect the heart development of zebrafish. The whole mount in situ hybridization was used to detect the regulation of nulpl on early and late stages of heart development of zebrafish. In situ hybridization was used to detect whether knocking out nulpl to affect heart development of zebrafish at the early or late stages. The results show that the heart has emerged as larger atrial and smaller ventricular and the effects is consistent with the previous conclusion. These results suggest that nulpl may affect the ventricular and atrial morphology of the zebrafish heart, respectively.