| Summary: | 碩士 === 國立中山大學 === 生物科學系研究所 === 107 === Genetic programs and signaling pathways are required for proper growth and patterning of blood vessels. Zebrafish is a powerful vertebrate model organism to study genetic control vascular development. In our previous study, transcription factors islet2 and coupTFIb regulated vein and ISV (intersegmental vessel) formation. Microarray analysis showed that some GTPase related genes are changed in expression in isl2/coupTFIb MO. GTPase signaling has been shown important for angiogenesis, migration, proliferation etc. Here, we reported two novel GTPase related genes ect2 and wasf1 that control vascular development in zebrafish. In-situ hybridization shows that ect2 and wasf1 mRNA is expressed in developing vessels, suggested the roles in vasculature. Loss of ect2 and wasf1 by morpholino knockdown individually impairs the growth of ISV and CVP (caudal vein plexus), suggesting the role of ect2 and wasf1 in controlling ISV and CVP growth. To address whether the cell death contributes to the ISV and CVP defects in wasf1 MO and ect2 MO, we performed TUNEL assay and AO staining. The data showed that vascular defects do not caused by cell death, but likely due to the impairment of proliferation and migration. To test molecular mechanisms of vascular defects in wasf1 MO and ect2 MO, we examined the expression of vascular markers and we found the remodeling the expression of vascular markers in wasf1 MO and ect2 MO. We also revealed the relationship between those GTPase related genes and Notch signals. To test molecular mechanisms between VEGF signals, BMP signals and ect2, wasf1, we performed western blot. The data showed that VEGF and BMP signals is regulated by ect2 and wasf1. Together, we showed that GTPase related proteins wasf1 and ect2 play important roles for vascular development in zebrafish.
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