Antizyme inhibitor decreases the cellular insults-interfered with zebrafish embryonic development

• Main Authors: Yuen-Chien, 陳岳謙
• Other Authors: 劉光耀
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/47153035634836416289

碩士 === 中山醫學大學 === 醫學研究所 === 97 === BACKGROUD: Antizyme inhibitor (AZI) delicately regulates ornithine decarboxylase (ODC) enzyme activity and polyamine transport. ODC plays a numerous roles in molecular and cellular functions, including developmental regulation, cell cycle, proliferation, cell death, differentiation and tumorigenesis. AZI has similar sequence and molecular weight with ODC. AZI, which has higher binding affinity to antizyme (AZ) more than ODC, can rescue ODC from AZ inhibition. The developmental mechanisms of zebrafish are similar with mammalian, highly. They are tiny size, easy feeding, short generation period, big limpid eggs, and developmental organs observation easily. However, how does AZI participate in the zebrafish is still unidentified. Therefore, we evaluate the effect of AZI on cellular insults during zebrafish embryonic development. METHODS: Cellular insults such as UVB, H2O2 and CQ were treated in zebrafish embryonic development. AZI expression was enhanced by microinjection of AZI DNA or protein. The phenotypes were observed and photographed with a microscope equipped with a digital camera. TUNEL assay was applied to identify apoptosis. The expressions of AZI, ODC and AZ mRNA were analyzed by using RT-PCR. RESULTS: We constructed the experiment model, zebrafish. We shown that UVB, H2O2 and CQ induced zebrafish embryonic development abnormal: malformation (similar defects of brain heart, development and heart function was abnormal, yolk sac enlargement and contortion of trunk and tail), increased death rate, decreased hatch rate and apoptotic level raise. Furthermore, AZI expression was reduced in cellular insults. The overexpression of AZI could recover the injuries as mention above. Beside, AZI shRNA leads malformation and increase death rate in zebrafish. CONCLUSIONS: According to these data, we suggest that AZI could resist stress and recover injury in zebrafish embryonic development.