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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 朱家瑩(Chia-Ying Chu) | |
| dc.contributor.author | Yen-Han Wang | en |
| dc.contributor.author | 王嬿涵 | zh_TW |
| dc.date.accessioned | 2021-05-19T17:55:51Z | - |
| dc.date.available | 2022-06-23 | |
| dc.date.available | 2021-05-19T17:55:51Z | - |
| dc.date.copyright | 2016-08-26 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-08-20 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7848 | - |
| dc.description.abstract | 再生是生物體更新或修復受損的細胞和組織，藉以維持正常生理功能與生存的方式。扁形動物渦蟲綱具有強大的再生能力，其體內存在大量多功能性成體幹細胞（neoblasts），能夠增生並分化成所有類型的細胞。因此，維持成體幹細胞增生與分化的平衡，對於渦蟲的生理恆定和再生過程都極為重要。DDX6蛋白屬於DEAD-box RNA解旋酶家族，真核生物中的DDX6同源蛋白不論在序列或功能上都具有高度保守性，並參與多項細胞生理機制。最近的研究更指出，DDX6能夠協同微核糖核酸（microRNA）影響哺乳類神經幹細胞分化，亦可透過降解mRNA和抑制轉譯的方式調控哺乳類上皮幹細胞的增生與分化。然而，對於渦蟲DDX6同源蛋白DjCBC-1是否能調控成體幹細胞增生與分化至今仍尚未明瞭。先前研究顯示，Djcbc-1表現在成體幹細胞、神經和生殖細胞當中，而抑制Djcbc-1的表現量會限制渦蟲眼點的再生。本研究進一步證實Djcbc-1高量表現於再生組織，並發現Djcbc-1的表現與否能顯著地影響處於再生期的渦蟲之存活率。除此之外，抑制Djcbc-1的表現量也會改變渦蟲腦部結構的完整性，並且大幅減弱渦蟲的負趨光性。我的研究結果顯示，Djcbc-1參與渦蟲個體的生理恆定、再生過程、以及負趨光性的調控。 | zh_TW |
| dc.description.abstract | Regeneration is the process of renewal and regrowth of damaged tissue to maintain the normal physiological functions for surviving. Planarian has a robust ability of regeneration. Regeneration in planarians requires neoblasts, the pluripotent somatic stem cells, which can proliferate and differentiate into all different cell types. Regulation of neoblasts proliferation and differentiation is critical for planarians homeostasis and regeneration. DDX6, a member of the DEAD-box protein family, is highly conserved among most eukaryotes. DDX6 homologs play multiple roles in many biological processes. Recent studies showed that DDX6 regulates the differentiation of mammalian neural stem cells by microRNAs, as well as regulates the proliferation and the differentiation of mammalian epidermal stem cells by mRNA degradation and translational repression. However, it is unclear that whether DjCBC-1, the DDX6 ortholog in planarians, also regulates the proliferation and differentiation of neoblasts. Previous studies showed that Djcbc-1 is expressed in neoblasts, neurons, and germline cells. In addition, depletion of Djcbc-1 limited the formation of photoreceptors. Here, I further demonstrated that Djcbc-1 is highly expressed in the regenerating tissue, and is essential for planarian survival at regeneration status. Besides, depletion of Djcbc-1 perturbed the homeostasis of brain structure and reduced the negative phototaxis in planarians. Collectively, these results suggest that Djcbc-1 participates in tissue homeostasis, regeneration, and the regulation of negative phototaxis in planarians. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-19T17:55:51Z (GMT). No. of bitstreams: 1 ntu-105-R03b21012-1.pdf: 5212717 bytes, checksum: 4982943a768a10dc36eb043c60b530ba (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 口試委員審定書 ... i
誌謝 ... ii 摘要 ... iii Abstract ... iv 1. Introduction ... 1 1.1 Regeneration in planarians ... 1 1.2 Neoblasts in planarian regeneration and homeostasis ... 2 1.3 Nervous systems and the behaviors of planarians ... 3 1.4 DEAD-box helicase 6 (DDX6) ... 5 1.5 DjCBC-1, the DDX6 ortholog in Dugesia japonica ... 7 2. Materials and Methods ... 11 2.1 Animals ... 11 2.2 Synthesis and purification of digoxigenin (DIG)-labeled RNA probes ... 11 2.3 Whole-mount in situ hybridization (WISH) ... 13 2.4 Gamma (γ)-irradiation ... 14 2.5 RNA isolation and cDNA preparation ... 14 2.6 Quantitative reverse-transcription PCR (RT-qPCR) ... 15 2.7 Fluorescence-activated cell sorting (FACS) ... 16 2.8 DsRNA-mediated RNA interference (RNAi) experiment ... 17 2.9 Photophobia assay ... 18 2.10 Motility assay ... 19 2.11 Immunofluorescence (IF) staining with phospho-Histone 3 (H3p) (Ser10) or SYNORF1 (Synapsin) antibody ... 19 3. Results ... 21 3.1 Djcbc-1 was highly expressed in brain and neoblasts of planarian ... 21 3.2 Djcbc-1 was highly expressed in the regenerating tissue of planarian ... 22 3.3 Djcbc-1 was essential for the survival of newly regenerated planarian ... 23 3.4 Depletion of Djcbc-1 delayed the eyespots formation during planarian head regeneration ... 25 3.5 The neoblasts population was not declined after Djcbc-1 RNAi ... 25 3.6 Depletion of Djcbc-1 may affect the activity of neoblasts ... 26 3.7 Depletion of Djcbc-1 perturbed the brain of planarian during homeostasis ... 27 3.8 Depletion of Djcbc-1 impaired the brain regeneration of planarian ... 29 3.9 Depletion of Djcbc-1 abolished the negative phototaxis of planarian ... 30 4. Discussion ... 32 4.1 Summary and significance ... 32 4.2 Depletion of Djcbc-1 in newly regenerated planarians results in abnormal phenotype, even leads to death ... 34 4.3 Planarian behavior and the subtype neurons of CNS ... 36 4.4 The function of DjCBC-1 in neurons ... 38 4.5 Perspective ... 38 5. Figures ... 40 Figure 1 Djcbc-1 was highly expressed in brain and neoblasts of planarian ... 40 Figure 2 The expression level of Djcbc-1 was decreased after γ-irradiation ... 41 Figure 3 Djcbc-1 was highly expressed in FACS isolated X1 and X2 populations from the body of planarian ... 42 Figure 4 Djcbc-1 was highly expressed in the regenerating tissue of planarian ... 44 Figure 5 Djcbc-1 RNAi was sufficient to deplete 80% of Djcbc-1 in planarian ... 46 Figure 6 Small planarians were more sensitive to Djcbc-1 RNAi ... 47 Figure 7 Newly regenerated planarians were more sensitive to Djcbc-1 RNAi ... 49 Figure 8 Djcbc-1 was essential for the survival of newly regenerated planarian ... 50 Figure 9 Depletion of Djcbc-1 delayed the eyespots formation during planarian head regeneration ... 52 Figure 10 The expression patterns and levels of neoblasts-related marker genes after Djcbc-1 RNAi ... 54 Figure 11 Depletion of Djcbc-1 increased the relative amounts of isolated X1 but not the X2 cells ... 55 Figure 12 Depletion of Djcbc-1 increased the mitotic cells of planarians ... 57 Figure 13 Djcbc-1-depleted planarians were more sensitive to γ-irradiation with the dosage of 5 Gy or 90 Gy ... 58 Figure 14 Depletion of Djcbc-1 had no effect on the planarian eyespots during homeostasis ... 59 Figure 15 Depletion of Djcbc-1 perturbed the planarian brain during homeostasis ... 60 Figure 16 Gene expression levels of FACS-separated cell populations in Djcbc-1-depleted planarian ... 62 Figure 17 Depletion of Djcbc-1 did not impair the eyespots regeneration of planarian ... 63 Figure 18 Depletion of Djcbc-1 impaired the brain regeneration of planarian ... 64 Figure 19 Depletion of Djcbc-1 abolished the negative phototaxis of planarian... 66 Figure 20 Depletion of Djcbc-1 did not affect the planarian motility and the stimulating-escape response ... 68 Figure 21 Summary of the function of DjCBC-1 in planarian ... 69 6. Reference ... 71 | |
| dc.language.iso | en | |
| dc.title | 渦蟲DDX6核糖核酸解旋酶DjCBC-1之功能研究 | zh_TW |
| dc.title | Functional study of the planarian RNA helicase DDX6, DjCBC-1 | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 郭典翰(Dian-Han Kuo),陳示國(Shih-Kuo Chen) | |
| dc.subject.keyword | DEAD-box核糖核酸解璇?,DDX6,DjCBC-1,組織恆定性,再生,渦蟲, | zh_TW |
| dc.subject.keyword | DEAD-box helicase 6,DDX6,DjCBC-1,homeostasis,regeneration,planarian, | en |
| dc.relation.page | 81 | |
| dc.identifier.doi | 10.6342/NTU201602791 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2016-08-21 | |
| 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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