碩士 / 國立彰化師範大學 / 生物技術研究所 / 96 / Copper (Cu) is an essential trace element for all organisms, but excess copper exposure can exert a variety of pathological effects in fish. In this study, we treated zebrafish embryos with 2.5-10 ppm either Cu+ or Cu ++ for 48h, survival rate was decreased and abnormal embryos was increased significantly. Lethal Concentration Fifty (LC50) was 5 ppm for either Cu+ or Cu++ treated. Reactive oxygen species (ROS) of embryos under copper ions treated were elevated by DCFH-DA analysis. The levels of protein carbonylation and lipid peroxidation were increased significantly after Cu++ treatment. We have evaluated the five antioxidant enzyme gene expression under 5 ppm copper treated by Real-time RT-PCR. The results showed that the expressions of Cu/Zn-SOD, Mn-SOD, Catalase, peroxiredoxin 2 and peroxiredoxin 6 gene were increased at 6 h, but Mn-SOD, peroxiredoxin 1 and peroxiredoxin 6 decreased at 48 h. In Mn-SOD promoter assays, we identified that MTF-1 was an important transcription factor to Mn-SOD gene expression under copper treated. Using morpholino to knock down Mn-SOD gene expression at one-cell stage of embryonic development, the embryo’s ROS and lipid peroxidation were increased the teratogenesis, especially at heart failure. The results showed that Mn-SOD was important for zebrafish embryonic development and oxidative stress resist.
| Identifer | oai:union.ndltd.org:TW/096NCUE5108004 |
| Date | January 2008 |
| Creators | Chin-Yu Cheng, 鄭進裕 |
| Contributors | Chuian-Fu Ken, 耿全福 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 168 |
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