Cellular and Molecular Study in Thioacetamide Induced Edema in Pericardial Sac during Zebrafish Development

• Main Authors: Po- Rung Jiang, 江柏蓉
• Other Authors: Chuian - Fu Ken, Ph.D.
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/57131851557503070487

碩士 === 國立彰化師範大學 === 生物技術研究所 === 101 === Thioacetamide (TAA) is a hepatotoxin that can cause hepatocellular carcinoma in mice, but little is known about its toxic effect on embryonic development. In the previous study, we treated zebrafish embryos with 0-32 mM TAA for 0-72 h, the survival rate were decreased to about 80% and the edema in pericardial sac (eps) were shown. For understanding the mechanism about TAA teratogenesis, we used DNA microarray to analyze the gene expression in zebrafish embryos when responses to TAA treatment. After data mining, there are 46 genes associated with organogenesis, and 107 genes involved in regulation of transcription. Among these genes, nrp2b and mef2cb were down-regulated to 2 or 1.8 fold after TAA treatment. nrp2b is a receptor that VEGF-mediated vascular development and mef2cb as a novel regulator in heart development. We then test the TAA downregulated nrp2b and mef2cb that can cause the edema in pericardial sac. 32 mM TAA treated embryos showed significantly decreased nrp2b and mef2cb gene expression, knock-down either nrp2b or mef2cb caused 55% and 56% eps on 24hpf, the results were the same as TAA treated embryos. Our laboratory has demonstrated previously the TAA induce embryonic a lot of produce H2O2. Therefore, we use 4 mM H2O2. treatment embryonic 6-24 hours then of quantitative RT-PCR and real-time qPCR, significantly reduce the more than 2 fold also caused a increase in embryonic eps the embryo mef2cb nrp2b amount of gene expression and increased mortality rate. Embryos processing 4mM H2O2 to 60 phf will not only cause eps makes Mast Cell aggregation in the pericardial sac near。By the above results, you can see that the TAA induce embryonic large number of generated the H2O2 that cuase reduce nrp2b and mef2cb gene expression and lead to eps eventually leading to death.