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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 盧國賢 | |
| dc.contributor.author | Yen-Chia Lee | en |
| dc.contributor.author | 李妍葭 | zh_TW |
| dc.date.accessioned | 2021-06-08T00:27:32Z | - |
| dc.date.copyright | 2013-09-24 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-07-10 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17646 | - |
| dc.description.abstract | Introduction:Arecoline, the major alkaloid in the betel nut, has been reported to be potent in inducing developmentally toxic effects by generally lowering the embryo weight and retarding development of the embryo. Zebrafish has been suggested as a model organism for vertebrate development, due to its rapid extracorporeal development, the transparent embryo, easy maintenance, and availability of gene markers. We explore the effect of arecoline on the skeletal muscle development of zebrafish.
Material and Methods:Arecoline was administered to zebrafish embryos by incubation at concentrations ranging from 0.001–0.04 %, treating from 4 to 24 hpf (hours post fertilization). We recorded the morphological changes, survival rates, and the body length, and applied immunohistochemistry (IHC), confocal microscopy, and conventional light electron microscopy to investigate the morphological changes of zebrafish skeletal muscle tissue. Results:The survival rate of arecoline-treated embryos significantly declined as the arecoline concentration increased, the treated embryos also showed general growth retardation and the swimming (motor) ability impairments. Immunohistochemistry (IHC) and microscopic studies demonstrated that major phenotypical changes are the myosin heavy chain in a relatively looser arrangement. Ultrastructural observation revealed an alteration of myofibrils arrangement and the swelling of mitochondria. RT-PCR analyse of mitochondrial function genes showed that at 48 and 96 hpf, the expression of the genes including sdhb, coxI, cox4i1, atp5a1, and atp5f1 significantly declined as the arecoline concentration increased, but the expression of mt_cytb increased. Conclusions:We confirmed that arecoline causes general growth retardation, skeletal muscle impairment, mitochondrial structure damage and the alteration of mitochondria functional genes expression in zebrafish embryo. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T00:27:32Z (GMT). No. of bitstreams: 1 ntu-102-R98446010-1.pdf: 6150686 bytes, checksum: 970f98d1b5887823ec9db19800e903db (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 中文摘要…………………………………………………………………………………………1
英文摘要…………………………………………………………………………………………2 縮寫表……………………………………………………………………………………………4 一、緒言（Introduction）………………………………………………5 1.1 前言……………………………………………………………………………………5 1.2 檳榔子………………………………………………………………………………5 1.3 檳榔鹼(Arecoline）…………………………………………………6 1.4 檳榔鹼造成的胚胎畸形…………………………………………………6 1.5 斑馬魚Zebrafish（Danio rerio）…………………7 1.5.1 斑馬魚作為動物模式在本研究之應用…………………7 1.6 檳榔鹼及粒線體疾病………………………………………………………8 二、目的（Purposes）…………………………………………………………9 三、材料與方法(Materials and methods)……………10 3.1化學藥品………………………………………………………………………………10 3.1.1一般化學藥品…………………………………………………………………10 3.1.2 Danieau buffer……………………………………………………10 3.1.3 Danieau buffer with 1x PTU……………………11 3.2 斑馬魚之飼養與受精卵之收集………………………………………11 3.3 斑馬魚胚胎之培養……………………………………………………………11 3.4 檳榔鹼之處理……………………………………………………………………11 3.5 胚胎之觀察紀錄及照相……………………………………………………12 3.6 數據圖表之統計整理…………………………………………………………12 3.7免疫組織化學染色…………………………………………………………………12 3.7.1 斑馬魚胚胎之固定…………………………………………………………12 3.7.2 實驗步驟……………………………………………………………………………13 3.7.3 全覆式包埋及共軛焦顯微拍照………………………………………13 3.8 電子顯微鏡標本製備……………………………………………………………13 3.9 反轉錄聚合酶鏈鎖反應（RT-PCR）………………………………14 四、結果（Results）…………………………………………………………………15 4.1 斑馬魚胚胎經檳榔鹼處理後之存活率………………………………15 4.2 斑馬魚胚胎經檳榔鹼處理後之形態差異…………………………15 4.3 斑馬魚胚胎經檳榔鹼處理後之長度統計分析…………………16 4.4 檳榔鹼處理對斑馬魚胚胎運動功能的影響………………………17 4.5 免疫組織化學染色…………………………………………………………………17 4.6 一般光學顯微鏡觀察……………………………………………………………18 4.7 電子顯微鏡觀察……………………………………………………………………18 4.8 反轉錄聚合酶鏈鎖反應（RT-PCR）………………………………19 五、討論（Discussion）…………………………………………………………20 5.1 檳榔鹼對胚胎發育的影響……………………………………………………20 5.2 檳榔鹼對斑馬魚胚胎發育的影響………………………………………20 5.3 檳榔鹼對斑馬魚胚胎運動功能的影響………………………………21 5.4 檳榔鹼對骨骼肌發育的影響…………………………………………………22 5.4.1 免疫螢光染色與一般光學顯微鏡之觀察………………………22 5.4.2 一般電子顯微鏡之觀察（超微結構）…………………………23 5.5 檳榔鹼對粒線體的影響…………………………………………………………24 六、結論（Conclusions）………………………………………………………26 參考文獻……………………………………………………………………………………………27 圖表說明……………………………………………………………………………………………35 | |
| dc.language.iso | zh-TW | |
| dc.title | 檳榔鹼對斑馬魚胚胎發育的影響 | zh_TW |
| dc.title | Effect of Arecoline on the Embryonic Development of Zebrafish (Danio rerio)- With special reference to skeletal muscle | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 周逸鵬,周?珠,龔秀妮,張綺芬 | |
| dc.subject.keyword | 檳榔鹼,斑馬魚,骨骼肌,粒線體, | zh_TW |
| dc.subject.keyword | Arecoline,Zebrafish,skeletal muscle,mitochondria, | en |
| dc.relation.page | 54 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2013-07-10 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 解剖學暨生物細胞學研究所 | zh_TW |
| 顯示於系所單位： | 解剖學暨細胞生物學科所 | |
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