碩士 / 國立臺灣海洋大學 / 環境生物與漁業科學學系 / 95 / Abstract
The zebrafish (Danio rerio) is vertebrate. It has common character with human in cardiovascular, immunity and vision system. Because it is easy to observe embryo development, the life of cycle is short and it is easy to microinjection. It is an important model in every toxicity experiment and it is to valuate danger environment. The TBT (tributyltin）is the permanent organic pollution in the environment and it can cause many aminals’ physiology and morphology change.
The purpose of this study is to understand TBT toxicity to zebrafish and its liver. Concentrations of TBT used for experiment groups are 0 ug/l , 5 ug/l , 25 ug/l , 50 ug/l , 75 ug/l and 100 ug/l while control group is 0 ug/l. There are 12 individuals of zebrafish used for each group during 24 hours exposed time. Three repeat for each experiment has been done.
The dead fish number is recorded for each experiment. And when experiment completed, we collected zebrafish fresh livers and take out their tissue to slice and HE stain. Further, we expose zebrafish liver cell line in different concentration of TBT, and record its survival rate by fluorescent stain and DNA electrophosis picture to determine whether the ZF-L cell is necrotic or apoptotic.
Results of acute toxicity show that TBT concentration of 24hr-LC50 is 46 ug/l. In the end of acute toxicity experiment, fish’s liver is taken out to examine its morphology. It can be found that liver become tumescent, and color deterioration.
With the paraffin-embedded tissue, immersing HE stain, it can be observed that liver damage condition in different concentration of TBT. After the ZF-L exposed with TBT up to 6 hrs and 24 hrs, trypan blue is used to calculate live cells and dead cells. It shows that there is no significant different cytotoxicity. By using fluorescent stain, it shows that cell become red dependent on TBT concentration. Basing DNA electrophoresis picture, the DNA are fragments. When use Western Blot analysis Bax protein and Bcl-2 protein, it shows that Bax protein to increase and Bcl-2 protein to reduce, dependent on TBT concentration.
In conclusion, hope these results are referable to the study of gene signal pathway of liver cell apoptosis. As well as, hope these results can let some people to understand the TBT toxicity to fish’s liver. Therefore, if TBT accumulates in human’s liver, the toxicity can not be neglected.
Key word：Tributyltin, Acute toxicity, ZF-L, Apoptosis, Zebrafish (Danio rerio).
| Identifer | oai:union.ndltd.org:TW/095NTOU5451007 |
| Date | January 2007 |
| Creators | Pao-Ying Kuo, 郭寶瓔 |
| Contributors | Chun-Te Chen, 陳俊德 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 83 |
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