an in vivo method to quantify lymphangiogenesis in zebrafish在斑马鱼体内方法量化lymphangiogenesis.pdfVIP

An In Vivo Method to Quantify Lymphangiogenesis in Zebrafish 1 1 2 1 1 2 Scott J. Hoffman , Peter J. Psaltis , Karl J. Clark , Daniel B. Spoon , Colin D. Chue , Stephen C. Ekker , Robert D. Simari1,2* 1 Division of Cardiovascular Diseases, Mayo Clinic, Rochester, Minnesota, United States of America, 2 Division of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota, United States of America Abstract Background: Lymphangiogenesis is a highly regulated process involved in the pathogenesis of disease. Current in vivo models to assess lymphangiogenesis are largely unphysiologic. The zebrafish is a powerful model system for studying development, due to its rapid growth and transparency during early stages of life. Identification of a network of trunk lymphatic capillaries in zebrafish provides an opportunity to quantify lymphatic growth in vivo. Methods and Results: Late-phase microangiography was used to detect trunk lymphatic capillaries in zebrafish 2- and 3- days post-fertilization. Using this approach, real-time changes in lymphatic capillary development were measured in response to modulators of lymphangiogenesis. Recombinant human vascular endothelial growth factor (VEGF)-C added directly to the zebrafish aqueous environment as well as human endothelial and mouse melanoma cell transplantation resulted in increased lymphatic capillary growth, while morpholino-based knockdown of vegfc and chemical inhibitors of lymphangiogenesis added to the aqueous environment resulted in decreased lymphatic capillary growth. Conclusion: Lymphatic capillaries in embryonic and larval zebrafish can be quantifi