Effects of Alkaline phosphatases on Zebrafish Embryonic Hematopoiesis

• Main Authors: Wen-Chan, 陳文銓
• Other Authors: 謝錦源
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/46129662379418584919

碩士 === 中山醫學大學 === 生化暨生物科技研究所 === 94 === Alkaline phosphatase (AP), in the past study has been known three out of four ALP isozymes are tissue-specific - placental (PLAP), germ cell (GCAP), the third is intestinal (IAP), and the last is tissue non-specific (TNAP). The TNAP was involved into mineralization and calcification in skeleton. However, recent research showed that mice lacking TNAP die from seizures due to defective metabolism of vitamin B6, and reduced level of inhibitory neurotransmitter γ-aminobutyric acid (GABA) in the brain. ALP has been established as a marker enzyme for a differentiated blood/brain barrier (BBB) phenotype in vivo and in vitro. It is known that TNAP existed in zebrafish, and expressed at early developmental stages of zebrafish embryos. Thus zebrafish as animal was selected as model to study the role of TNAP in zebrafish embryo development process. In this study, we analyze spatiotemporal expression of zebrafish TNAP by RT-PCR and whole-mount in situ hybridization. And the siRNA or morpholino of zebrafish TNAP were microinjected into zebrafish one-cell embryos to observe the roles of TNAP on embryogenesis. It was found that after TNAP knockdown embryos were delayed in embryogenesis, bended of notochord in larva period and decreased hematopoiesis (by o-dianisidnie staining) in 48hpf. In the future, we will use specific RNA probe to analyze the role of TNAP in hematopoiesis at zebrafish early development stage.