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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 朱家瑩(Chia-Ying Chu) | |
| dc.contributor.author | Chao-I Chuang | en |
| dc.contributor.author | 莊昭儀 | zh_TW |
| dc.date.accessioned | 2021-06-08T01:07:03Z | - |
| dc.date.copyright | 2014-09-02 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-19 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18474 | - |
| dc.description.abstract | 細胞自噬是一種可以將細胞中不需要或者是失去正常功能的組成物降解的基礎代謝路徑，不論是從酵母菌到人類，細胞自噬都具有高度保留性。近期研究指出細胞自噬可能參與調控幹細胞在老化以及再生、組織重組的恆定性。在人類細胞的研究中也顯示成體幹細胞的自我更新以及多元性分化必須要經歷細胞自噬的過程。儘管如此，細胞自噬在渦蟲幹細胞以及再生上的功能仍未經仔細探討。
為了瞭解細胞自噬在渦蟲再生中所扮演的角色，我從本實驗室建構的東洋渦蟲轉錄子資料庫中找到一些酵母菌細胞自噬相關因子(Atgs)的同源性基因。其中我們選殖了三個酵母菌Atg8同源基因，並分別命名為DjAtg8A 、DjAtg8B以及DjAtg8C。在針對此三基因的原位雜交實驗結果中顯示，這些基因表現於渦蟲腸道中，顯示這些基因在細胞內吞作用和溶酶體代謝路徑上可能具有功能上的保留性。在酵母菌以及HeLa細胞的功能替代檢測中也顯示Atg8同源基因的保留性。此外加碼射線的照射會使得這些DjAtg8基因的表現量上升，更進一步地使得DjAtg8A脂化，這些結果顯示渦蟲的細胞自噬路徑一樣會因環境壓力而活化並且活化路徑也與其他模式物種相近。此外，當我們以RNAi方式降低渦蟲體內不同DjAtg8基因的表現後，去除DjAtg8B會造成渦蟲斷裂並死亡。這些結果提供證據使我們判斷細胞自噬在渦蟲消化系統以及對環境壓力反應上的功能，但是在渦蟲再生的功能仍需近一步釐清。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-08T01:07:03Z (GMT). No. of bitstreams: 1 ntu-103-R01b41022-1.pdf: 4029867 bytes, checksum: 867e1627bb06bdece3065b6ca7e64767 (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 口試委員審定書 i
致謝 ii 摘要 iii Abstract iv 1. Introduction 1 1.1 Autophagy 1 1.2 Autophagy and stem cells 4 1.3 Regeneration and remodeling of planarian 6 1.4 Autophagy in planarian 9 1.5 Exploring the Atg8-dependent autophagy in Dugesia japonica 10 2. Material and Methods 11 2.1 Animals 11 2.2 Cloning of Dugesia japonica Atg8 complementary DNA 11 2.3 Quantitative reverse-transcription PCR (RT-qPCR) analysis of gene expression levels 12 2.4 Whole mount in situ hybridizations 13 2.5 Cell culture and transient expression of YFP fusion protein 14 2.6 Western blotting of HeLa cells 15 2.7 Western blotting of yeast cells 16 2.8 Yeast strains and medium 16 2.9 Ape1 maturation assay of yeast 17 2.10 RNA interference (RNAi) experiments 17 3. Results 18 3.1 Cloning of the Dugesia Japonica Atg8 paralogs 18 3.2 Expression pattern of DjAtg8 in Dugesia japonica 19 3.3 Functional conservation of DjAtg8 paralogs in mammalian cells 20 3.4 DjAtg8 partially participates in autophagy and Cvt pathway in yeast 21 3.5 Autophagy is up-regulated by stress in planarian 23 3.6 Expression of DjAtg8-paralogs during regeneration 24 3.7 Phenotypes in DjAtg8s-depleted planarians 26 3.8 Identification of planarian genes in Atg8 conjugation systems 27 4. Discussions 29 4.1 DjAtg8A, DjAtg8B and DjAtg8C are members of Atg8 family 29 4.2 DjAtg8A, DjAtg8B and DjAtg8C partially compensates the function of autophagy in yeast and promote autophagy in planarian 33 4.3 DjAtg8 paralogs may not play roles in neoblasts 36 4.4 The DjAtg8 paralogs acts differently but all participate in autophagy in planarian 37 Figures 44 Supplementary data 63 | |
| dc.language.iso | en | |
| dc.title | 渦蟲體內Atg8 蛋白之功能性研究 | zh_TW |
| dc.title | Identification and Characterization of Autophagy-Related Genes Atg8 in Planarian | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳俊宏(Jiun-Hong Chen),黃偉邦(Wei-Pang Huang) | |
| dc.subject.keyword | 渦蟲,Atg8,細胞自噬,再生, | zh_TW |
| dc.subject.keyword | Planarian,Atg8,autophagy,regeneration, | en |
| dc.relation.page | 71 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2014-08-19 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生命科學系 | zh_TW |
| 顯示於系所單位： | 生命科學系 | |
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