| Summary: | 碩士 === 慈濟大學 === 人類遺傳研究所 === 94 === The importance of fibroblast growth factor (FGF) signals in liver organogenesis has been reported in mammals. We screened the expression of FGF receptors (FGFR) in three developmental stages of zebrafish liver and found the FGFR2b was the major subtype. We also found FGF10, one of the ligands of the FGFR2b, expressed at high level in early liver development. However, the roles of FGF10/FGFR2 in liver development of zebrafish are not clear. We knocked down the expression of FGF10 by injecting antisense morpholino (MO) into the embryos of liver type fatty acid binding protein (LFABP)-GFP transgenic fish. The results indicated that the liver of FGF10 and FGFR2 morphants were smaller than wild-type, especially in FGF10/FGFR2 double morphants. Using whole-mount in situ hybridization, the expression pattern of liver specific gene LFABP was reduced in FGFR2 morphant and double morphant. The expression of downstream target gene pea3 was decreased in the morphants, especially in FGF10 morphants. The daedalus (dae) zebrafish, in which the FGF10 gene was disrupted, will crossed with the LF2.8-GFP transgenic fish to generate the dae/LFABP fish. The fish will be analyzed by direct observation of GFP expression in the liver and subsequent histological analysis. On the other hand, we used the liver-specific L-FABP promoter to ectopically overexpress FGF10 in the liver. We cloned FGF10 into the LF2.8-MCS-V5/CMV-RFP bi-functional vector and injected to embryos. The injected embryos with RFP will be analyzed for the expression of exogenous genes. In the future, we will create the stable transgenic fish lines with FGF10 overexpression.
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