碩士 / 國立臺灣海洋大學 / 生物科技研究所 / 99 / Zebrafish(Danio rerio) has been extensively used as a model organism for vertebrates and the alkylating agent ENU (N-ethyl-N-nitrosourea) is commonly used to generate a large scale of mutation because of its ability to cause DNA damages in spermatogonial stem cells. In this study, an ENU-generated zebrafish mutant(snake) was taken to examine whether changes in morphological features and defects in the gastrointestinal system had occurred. The eyes and heads of snake zebrafish at 4 days post fertilization(dpf) were obviously smaller than those of its sibling. More severe abnormal features, including curved body axes, withering of swim bladders and development defects in the digestive tract, were found to arise in 5 dpf snake organisms. Whole mount in situ hybridization showed no influence of ENU-induced mutation on the digestive tract when early genes(cp, hhex, prox1) specifically expressed in the liver, the pancreas and the intestinal tract were analyzed. In situ hybridization using a late liver-specific gene marker fabp10a indicated that a severe liver damage had occurred in 5 dpf snake mutant fish. Additionally, hybridization targeting the late expression gene trypsin showed a serious pancreas defect in 5 dpf snake zebrafish and the hybridization signals were barely detected in 6 dpf mutant fish, reflecting a complete loss of the exocrine function of the pancreas. Acridine orange(AO) staining revealed the presence of apoptotic cells in the heart and the head tissues of the mutant zebrafish. The results of this study demonstrated the sensitivity of the late development stages of liver and pancreas to ENU-induced gene mutations.
| Identifer | oai:union.ndltd.org:TW/099NTOU5613007 |
| Date | January 2011 |
| Creators | Yin-Shan Chang, 張銀珊 |
| Contributors | Guor-Mour Her, 何國牟 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 60 |
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