Abstract:

BACKGROUND: Previous studies found that in different periods of early development of zebrafish, embryos treated with certain concentration of deguelin show varying degrees of developmental delay and abnormal phenotype.
OBJECTIVE: To observe the effect of deguelin on expression of Wnt signaling pathway related genes in Zebrafish embryo during early embryo development.
METHODS: Zebrafish embryos that incubated to the shield period were collected and put into the 6-well plate, 0.6 μmol/L deguelin solution were added and treated in dark, and then cultured in the incubator under 28 ℃. Zebrafish embryos treated with 0.1% dimethylsulfoxide solution were as control group, and cultured in the same conditions. At 6, 8, 10, 12, 14 and 24 hours after fertilization, phenotype of Zebrafish embryos was observed with upright fluorescence microscope. The expression levels of target genes before and after treatment were tested by quantitative real time PCR.
RESULTS AND CONCLUSION: The embryos treated by deguelin on shield stage stop growing and stay at bud stage; the expressions of wnt9a, wnt10b, axin2 as well as snail1b and Jnk2 related with Wnt signal were changed significantly, among them, the expression of wnt9a, wnt10b, axin2 and snail 1b were decreased, while the expression of Jnk2 was increased. Deguelin has a negative effect on Zebrafish in vivo Wnt signal.

Key words: tissue construction, cytology experiment in tissue construction, deguelin, real-time PCR, Wnt signaling pathway, provincial grants-supported paper