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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李秀香(Hsiu-Hsiang Lee) | |
| dc.contributor.author | Jian-Ming Ke | en |
| dc.contributor.author | 柯建銘 | zh_TW |
| dc.date.accessioned | 2021-05-15T17:52:31Z | - |
| dc.date.available | 2020-03-12 | |
| dc.date.available | 2021-05-15T17:52:31Z | - |
| dc.date.copyright | 2015-03-12 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-12-04 | |
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(2013) A Cullin1-based SCF E3 ubiquitin ligase targets the InR/PI3K/TOR pathway to regulate neuronal pruning. PLoS Biol. 11(9):e1001657. Xiang Y, Yuan Q, Vogt N, Looger LL, Jan LY, Jan YN. (2010) Light-avoidance-mediating photoreceptors tile the Drosophila larval body wall. Nature 468(7326):921-6. Yoshikawa S, McKinnon RD, Kokel M, Thomas JB. (2003) Wnt-mediated axon guidance via the Drosophila Derailed receptor. Nature 422(6932):583-8. Zeng X, Tamai K, Doble B, Li S, Huang H, Habas R, Okamura H, Woodgett J, He X. (2005) A dual-kinase mechanism for Wnt co-receptor phosphorylation and activation. Nature 438(7069):873-7. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/5143 | - |
| dc.description.abstract | Neuronal remodeling plays a critical role during development from invertebrates to vertebrates. Pruning, one of the neuronal remodeling mechanisms, is a process to selectively remove specific parts of neurons without causing cell death. In Drosophila, the class IV dendritic arborization (da) neurons, which undergo pruning process to eliminate larval dendrites during metamorphosis, is an ideal model system to study the underlying mechanisms of pruning. Previous studies showed that ubiquitin proteasome system (UPS) initiates the dendrite pruning. To address the dynamics of proteasome activity during pruning process, we generated a photoconvertible reporter system. The decrease of converted fluorescent signals over a period of time could be used to monitor the proteasome activity in vivo. Wingless-Int (Wnt) signaling is crucial for variety of biological events. It could be divided into three major pathways: canonical Wnt pathway, planar cell polarity (PCP) pathway, and calcium pathway. Previous studies reported a trophic role for Wnt-Ror kinase signaling to regulate neuronal pruning in C.elegans, thus highlighted the regulatory roles of Wnt signaling. Here, we identified dishevelled (dsh), a Wnt signaling molecule, as a candidate to regulate Drosophila dendrite pruning. Dsh is a cytoplasmic phosphoprotein, which is involved in both canonical Wnt and PCP pathways. To determine which pathway of Wnt signaling can regulate dendrite pruning, we examined the dendrite pruning of class IV da neurons in various dsh mutant flies, and found the canonical Wnt pathway is associated with dendrite pruning. Next, we identified which Wnt receptors might act upstream of dsh during dendrite pruning. Consistent with the fact that the redundant roles of fz and fz2 in canonical Wnt signaling, both fz and fz2 loss of function mutants showed pruning defects. Furthermore, a co-receptor arrow, which is required for canonical Wnt signaling, also showed some dendrite severing defects when it lost its function. Finally, the canonical Wnt-specific nuclear TCF caused critical severing defects when expressing its N-terminal deletion form. Taken together, these results demonstrated that the canonical Wnt pathway is essential for Drosophila dendrite pruning. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-15T17:52:31Z (GMT). No. of bitstreams: 1 ntu-103-R01448014-1.pdf: 4534979 bytes, checksum: 71c1b40994e62cb7d64110024337f70c (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 口試委員會審定書……………………………………………………….i 誌謝…..………………………………………………………………… .ii Abstract………………………………………………………………....iii 摘要…..………………………………………………………………… .v Chapter 1. Introduction .……………………………………………...01 1. Neuronal remodeling............................................................................01 2. Drosophila class IV dendritic arborization (da) neurons……………..02 3. Drosophila dendrite pruning………………………………………….03 4. Ubiquitin-proteasome system (UPS)…………………………………04 5. Wnt pathways………………………………………………………...06 6. Dishevelled (dsh)……………………………………………………..08 7. Multiple Wnt receptors……………………………………………….10 8. Other molecules involved in Wnt signaling………………………….11 9. Hypothesis……………………………………………………………12 Chapter 2. Materials Methods………………………………..……15 Chapter 3. Results…………………………………………..………….18 1. The relationship between the dynamics of proteasome activity and Drosophila dendrite pruning…………………………………………18 1.1 Knockdown of proteasome subunits in class IV da neurons caused pruning defects……………………...……………………………….18 1.2 The proteasome activities of Ub-Dendra2 and Dendra2 control in WT ddaC neurons during the larval stage………………………..………20 1.3 The proteasome activity of Ub-Dendra2 under knockdown of DTS5 and UAS-lacZ control backgrounds during the larval stage…...……21 2. Loss-of-function analysis of dishevelled in class IV da neurons.……22 2.1 The dsh mutants showed minor dendrite severing defects………….22 2.2 The genetic interaction between dsh and three identified regulators (Ik2, kat-60L1, and Spn-F) in class IV da neurons...………………..23 2.3 Mutational analysis of dsh with distinct mutated domains showed various morphological changes of dendrite pruning defects...………24 3. Loss-of-function analysis of multiple Wnt receptors in class IV da neurons…………..…………………………………………………..26 3.1 Loss-of-function analysis of two redundant Wnt receptors, fz and fz2, showed minor dendrite severing defects…………………….………26 3.2 Other receptors involved in multiple Wnt pathways……….……….28 3.3 Mutational analysis of Wg co-receptor arrow showed some dendrite pruning defects…………………...………………………………….29 4. Loss-of-function analysis of other Wnt members downstream of dsh in class IV da neurons…………………………….…………….………30 4.1 Expression of various mutant forms of sgg caused critical severing defects…………………………………..........................……………30 4.2 Expression of N-terminal-deletion TCF mutant form caused critical severing defects………………………...……………………………31 Chapter 4. Discussion………………………………………………….33 1. The application of photoconvertible reporter system of the UPS……34 2. The mutational analysis of Dishevelled…………...………………….35 3. The roles of two redundant Wnt receptors, Frizzled and Frizzled2, as well as the co-receptor arrow……………………………….……….36 4. Possible mechanisms for Wnt signaling to regulate dendrite pruning…………………….…………………………………………38 5. Crosstalk between protein degradation and Wnt pathway in dendrite pruning………………………………………….……………………39 Chapter 5. References………………...……………………………….41 Tables……………………………………….…………………………..48 Figures………………………………………….………………………53 List of Tables Table 1. List of fly mutant stocks…………………………...…………..48 Table 2. List of fly RNAi stocks…………………………...……………51 Table 3. List of enhancer trap lines………………………………..…….52 List of Figures Figure 1. Knockdown of proteasome subunits in class IV da neurons caused dendrite pruning defect………………………...…………….53 Figure 2. Proteasome activities of Ub-Dendra2 in class IV da neurons during the larval stage……………………………………………….55 Figure 3. Knockdown of dsh caused critical dendrite pruning defect…..58 Figure 4. No severing defect were observed in dsh mutants……………60 Figure 5. Dsh hemizygous mutants caused minor severing defect……...61 Figure 6. The genetic interaction between dsh and three identified regulators (Ik2, kat-60L1, and Spn-F) in class IV da neurons………63 Figure 7. Mutational analysis of dsh with distinct mutated domains showed various morphological changes of dendrite pruning defects..................................................................................................65 Figure 8. Mutants of two redundant Wnt receptors, fz and fz2, showed little dendrite severing defects……………………………………….68 Figure 9. Single or double Knockdown of fz and fz2 showed little dendrite severing defects…………………………………………….70 Figure 10. Other receptors involved in multiple Wnt pathways………...72 Figure 11. Mutational analysis of Wg co-receptor arr showed some dendrite pruning defects……………………………………………..73 Figure 12. Expression of various mutant forms of sgg caused severe dendrite pruning defects………….………………………………….76 Figure 13. Expression of N-terminal-deletion TCF caused critical dendrite severing defects…………………………………………….78 | |
| dc.language.iso | en | |
| dc.subject | dishevelled | zh_TW |
| dc.subject | TCF | zh_TW |
| dc.subject | 第四樹突型神經元 | zh_TW |
| dc.subject | 蛋白?體活性 | zh_TW |
| dc.subject | 神經樹突修剪 | zh_TW |
| dc.subject | Wnt訊息傳導路徑 | zh_TW |
| dc.subject | Wnt受體 | zh_TW |
| dc.subject | dendrite pruning | en |
| dc.subject | TCF | en |
| dc.subject | Wnt receptors | en |
| dc.subject | dishevelled | en |
| dc.subject | Wnt pathway | en |
| dc.subject | proteasome activity | en |
| dc.subject | class IV da neurons | en |
| dc.title | 蛋白酶體活性與Wnt訊息傳導路徑調控果蠅神經元樹突修剪之研究 | zh_TW |
| dc.title | Roles of Proteasome Activity and Wnt Signaling Regulation during Neuronal Dendrite Pruning in Drosophila | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 徐立中(Li-Chung Hsu),皮海薇(Hai-Wei Pi) | |
| dc.subject.keyword | 神經樹突修剪,第四樹突型神經元,蛋白?體活性,Wnt訊息傳導路徑,Wnt受體,dishevelled,TCF, | zh_TW |
| dc.subject.keyword | dendrite pruning,class IV da neurons,proteasome activity,Wnt pathway,dishevelled,Wnt receptors,TCF, | en |
| dc.relation.page | 78 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2014-12-04 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
| 顯示於系所單位： | 分子醫學研究所 | |
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