Cloning, Expression And Functional Studies Of Zebrafish SEMA4C Gene

Sema4c is a trans-membrane protein of semaphorins that have been confirmed with axon guidance and myotube formation. In the early experiments, we have showed sema4c could enhance endothelial cells movement and tube formation in vitro, but it was unclear what kind of role sema4c plays in vivo.In order to explore the functions of sema4c in vivo, we applied an ideal model organism--zebrafish, and focused on sema4c of functions that regulate angiogenesis and lymphangiogenesis in zebrafish. In this study, we blasted gene sema4c from zebrafish gene database, which is termed zsema4c. By means of sequence analysis, protein comparison and homology analysis, we verified that zsema4c is an orthologous gene of human sema4c, and they are highly conserved in evolution. Whole-mount in situ hybridization assay showed that zsema4c was expressed maternally and ubiquitously during embryogenesis, implying a role in early embryonic development. Zsema4c expressed starting from one cell stage, specially distributed in blood vessels at 24hpf. The expression pattern is similar to temporal-spatial expression of ZFLT4 and ZVEGFC. Using a morpholino-antisense gene knockdown approach, we showed that almost all embryos arise vascular developmental disorders at high dose morpholino, the main phenotype is defect in branch vessels. However, morpholino injection at lower doses inhibited thoracic duct formation, it's only 30% embryo have this special phenotype.The lymphatic and blood vessels are closely related with cancer metastasis, we first confirmed that zsema4c play an important role in zebrafish angiogenesis and lymphangiogenesis, it maybe become a new target in clinical cancer treatment.