动物造模 z-bio 2022-07-26 304

　　Objective: To explore the role of ripply1 gene in the development of the early dorsal-ventral axis in zebrafish.

　　Methods: The zebrafish whole-mount in situ hybridization technique was used to reveal the expression pattern of ripply1 gene in the early embryonic development of zebrafish, and the ripply1 mRNA was injected at the 1-cell stage of embryos by microinjection technology to highly express Ripply1 protein, and the embryos were observed at the later stage. Changes of dorsal and ventral marker genes and embryo morphological changes. Tol2 transgenic technology was used to construct GFP transgenic fish driven by the ripple1 promoter.

　　RESULTS: The results of in situ hybridization showed that the ripply1 gene was specifically expressed at the embryo shield, which is the predetermined back, during the early gastrula stage of zebrafish. After high expression of ripply1, the expression range of dorsal marker genes was expanded in the embryo shield stage, and the expression of abdominal marker genes was weakened. At 24 hours after fertilization, the embryos showed a severe dorsalization phenotype: enlarged head, weakened abdominal yolk extension, and tail trunk and tail. The area was reduced, and some even formed a second body axis. The resulting transgenic fish revealed maternal expression of ripply1, and GFP driven 1200 bases upstream of the transcription start site could mimic the expression pattern of endogenous genes.

　　Conclusion: ripply1 may be involved in the development of the early dorsal-ventral axis in zebrafish embryos.