动物实验,斑马鱼早期胚胎 z-bio 2021-01-21 341

　　Objective: To investigate the role of ripply1 gene in the early development of zebrafish dorsoventral axis.

　　 Method: Using zebrafish whole cyst in situ hybridization technology, reveal the expression pattern of ribply1 gene in the early embryonic development of zebrafish, and use microinjection technology to inject ribply1 mRNA in the 1-cell stage of embryo. , Ripply1 and above proteins are highly expressed. Observe the embryonic dorsal and abdominal marker genes and embryo morphology changes. Tol2 transgenic technology was used to construct GFP transgenic fish driven by ribply1 promoter.

　　Result: The result of in situ hybridization showed that ribply1 gene was specifically expressed in embryo shielding stage in the early stage of embryo shielding in zebrafish stomach. After highly expressing ripply1, the expression range of the dorsal marker gene in the blastocyst stage of the blastocyst expanded, and the expression of the abdominal marker gene was weakened. 24 hours after fertilization, the embryo showed a severe dorsal phenotype: the head enlarged. However, in some cases, the yolk extension of the abdomen becomes weaker, the torso of the tail and the area of the tail decrease, and another separate axis is formed. The resulting transgenic fish revealed the expression of maternal ribply1, and the GFP-driven transcription start site 1200 bases upstream can mimic the expression pattern of endogenous genes.

　　 Conclusion: Ripply1 may be involved in the early dorsoventral axis of zebrafish embryos.