請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69663
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳俊宏(Jiun-Hong Chen) | |
dc.contributor.author | Sheridan Ke-wing Fok | en |
dc.contributor.author | 霍其嶸 | zh_TW |
dc.date.accessioned | 2021-06-17T03:22:54Z | - |
dc.date.available | 2019-07-06 | |
dc.date.copyright | 2018-07-06 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-06-15 | |
dc.identifier.citation | Reference
Agata K. (2003) Regeneration and gene regulation in planarians. Curr. Opin. Genet. Dev.13:492–496. Agata K, Nakajima E, Funayama N, Shibata N, Saito Y, Umesono Y. (2006 ) Two different evolutionary origins of stem cell systems and their molecular basis. Semin. Cell Dev. Biol. 2006;17:503–509 Agata K, Saito Y, Nakajima E. (2007) Unifying principles of regeneration I: epimorphosis versus morphallaxis. Dev. Growth Differ. 2007;49:73–78 Agata, Kiyokazu, and Yoshihiko Umesono. (2008) Brain Regeneration from Pluripotent Stem Cells in Planarian. Philosophical Transactions of the Royal Society B: Biological Sciences 363.1500 2071–2078. PMC. Web. 26 Dec. 2017. Almuedo-Castillo, María et al. (2014 ) JNK Controls the Onset of Mitosis in Planarian Stem Cells and Triggers Apoptotic Cell Death Required for Regeneration and Remodeling. Ed. A. Aziz Aboobaker. PLoS Genetics 10.6 (2014): e1004400. PMC. Web. 26 Dec. 2017. Bergmann, Andreas, and Hermann Steller. (2010) Apoptosis, Stem Cells, and Tissue Regeneration. Science signaling 3.145: re8–. Bergantinos, C., Corominas, M., Serras F. (2010) Cell death-induced regeneration in wing imaginal discs requires JNK signalling Development, 137, pp. 1169-1179 Bottger, A., Alexandrova O. (2007) Programmed cell death in Hydra Semin. Cancer Biol., 17, pp. 134-146 Chera, S., Ghila, L., Dobretz, K., Wenger, Y., Bauer, C., Buzgariu, W., Martinou, J.C., Galliot, B. (2009) Apoptotic cells provide an unexpected source of Wnt3 signaling to drive hydra head regeneration Dev. Cell, 17, pp. 279-289 Elmore, S. (2007) Apoptosis: a review of programmed cell death Toxicol. Pathol., 35, pp. 495-516 Fan, Y., Bergmann, A. (2008) Apoptosis-induced compensatory proliferation. The Cell is dead. Long live the Cell! Trends Cell Biol., 18, pp. 467-473 Fan, Y., Bergmann A. (2008) Distinct mechanisms of apoptosis-induced compensatory proliferation in proliferating and differentiating tissues in the Drosophila eye Dev. Cell, 14, pp. 399-410 Fire, A., Xu S., Montgomery, M.K., Kostas, S.A., Driver, S.E., Mello, C.C., Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 1998;391:806–810. doi: 10.1038/35888 Florentin, Anat, and Eli Arama. (2012 ) Caspase Levels and Execution Efficiencies Determine the Apoptotic Potential of the Cell. The Journal of Cell Biology 196.4: 513–527. PMC. Galliot, B. & Chera, S. (2010) The Hydra model: disclosing an apoptosis-driven generator of Wnt-based regeneration Trends Cell Biol., 20 , pp. 514-523 Galliot, B., Ghila, L. (2010) Cell plasticity in homeostasis and regeneration Mol. Reprod. Dev., 77, pp. 837-855 Galliot, B., Miljkovic-Licina, M., de Rosa, R., Chera, S. (2006) Hydra, a niche for cell and developmental plasticity Semin. Cell Dev. Biol., 17, pp. 492-502 Gauron, Carole et al. (2013) Sustained Production of ROS Triggers Compensatory Proliferation and Is Required for Regeneration to Proceed. Scientific Reports 3: 2084. PMC. Web. 26 Dec. 2017. Hannon, G.J. RNA interference. Nature. 2002;418:244–248. doi: 10.1038/418244a Huang, S.A., Mishina, Y.M., Liu,S., et al., (2009) Tankyrase inhibition stabilizes axin and antagonizes Wnt signaling Nature 461, 614–620 Huh, J.R., Guo, M., Hay, B.A. (2004) Compensatory proliferation induced by cell death in the Drosophila wing disc requires activity of the apical cell death caspase Dronc in a nonapoptotic role Curr. Biol., 14, pp. 1262-1266 Iglesisa, M., Almuedo-Catsillo, M., Aboobaker, A.A., Salo, E. (2011) Early planarian brain regeneration is independent of blastema polarity mediated by the Wnt/β-catenin pathway. Developmental Biology, 358 (1) , pp. 68-78.Kragl, M., Knapp, D., Kumar, S. (2006) Caspase function in programmed cell death. Cell Death and Differentiation (2006) 14, 32–43 (2007) Kyrylkova K., Kyryachenko S., Leid M., Kioussi C. (2012) Detection of Apoptosis by TUNEL Assay. In: Kioussi C. (eds) Odontogenesis. Methods in Molecular Biology (Methods and Protocols), vol 887. Humana Press Krysko, D.V., Vanden Berghe T., D'Herde, K., Vandenabeele P. (2008) Apoptosis and necrosis: detection, discrimination and phagocytosis Methods, 44 (2008), pp. 205-221 Nacu, E., Khattak, S., Maden, M., Epperlein, H.H., Tanaka E.M. (2009) Cells keep a memory of their tissue origin during axolotl limb regeneration Nature, 460, pp. 60-65 Li, F., Huang, Q., Chen, J., Peng, Y., Roop, D.R., Bedford, J.S., Li, C.Y. (2011 ) Apoptotic cells activate the “phoenix rising” pathway to promote wound healing and tissue regeneration Development. Jun;138(12):2417-27. doi: 10.1242/dev.060764. McIlwain D.R., Berger T., Mak T.W. (2013) Caspase functions in cell death and disease. Cold Spring Harb. Perspect. Biol. 5:a008656. Minina,E.A., Coll, N.S., Tuominen, H & Bozhkov, P.V. (2017) Metacaspases versus caspases in development and cell fate regulation Cell Death and Differentiation 24, 1314–1325 Nacu, E., Tanaka E.M. (2011) Limb regeneration: a new development? Annu. Rev. Cell Dev. Biol. 27:409-40 Orita Y., Nishizaki K., Sasaki J., Kanda S., Kimura N., Nomiya S., Yuen K., Masuda Y. (1999) Does TUNEL staining during peri- and post-natal development of the mouse inner ear indicate apoptosis? Acta Otolaryngol Suppl 540, 22–26. Pearson, B.J., Sánchez Alvarado A. (2010) A planarian p53 homolog regulates proliferation and self-renewal in adult stem cell lineages Development, 137, pp. 213-221 Petersen, C.P., Reddien, P.W. (2009) A wound-induced Wnt expression program controls planarian regeneration polarity Proc. Natl. Acad. Sci. USA, 106, pp. 17061-17066 Pellettieri, J. et al (2010 ) Cell Death and Tissue Remodeling in Planarian Regeneration. Developmental biology 338.1 (2010): 76–85. Reddien, P.W., Bermange, A.L., Murfitt, K.J., Jennings, J.R., Sánchez Alvarado, A. (2005). Identification of genes needed for regeneration, stem cell function, and tissue homeostasis by systematic gene perturbation in planaria Dev. Cell, 8, pp. 635-649 Rode H. D., Eisel D., Frost I. (2004) Apoptosis, cell death and cell proliferation. 3rd ed. London: Roche Applied Science. Ryoo, H.D., Gorenc, T., Steller, H. (2004) Apoptotic cells can induce compensatory cell proliferation through the JNK and the Wingless signaling pathways Dev. Cell, 7, pp. 491-501 Sánchez Alvarado, A. (2006) Planarian regeneration: its end is its beginning Cell, 124 (2006), pp. 241-245 Tamura, K., Ohgo, S., Yokoyama, H. (2010) Limb blastema cell: a stem cell for morphological regeneration Dev. Growth Differ., 52, pp. 89-99 Tanaka, E., Reddien, P. (2001) The Cellular Basis for Animal Regeneration Developmental Cell, 21 (1) , pp. 172-185. Tasaki, J., Shibata, N., Nishimura, O., Itomi, K., Tabata, Y., Son, F., Suzuki, N., Akari, R., Abe, M., Agata, K., Umesono, Y. (2011) ERK signaling controls blastema cell differentiation during planarian regeneration. Development. Jun;138(12):2417-27 Tseng, A.S., Adams, D.S., Qiu,D., Koustubhan,P., Levin, M. (2007) Apoptosis is required during early stages of tail regeneration in Xenopus laevis Dev. Biol., 301, pp. 62-69 Yokoyama, H. (2008) Initiation of limb regeneration: the critical steps for regenerative capacity Dev. Growth Differ., 50, pp. 13-22 Vlaskalin, T., Wong, C.J., Tsilfidis C. (2004) Growth and apoptosis during larval forelimb development and adult forelimb regeneration in the newt (Notophthalmus viridescens) Dev. Genes Evol., 214, pp. 423-431 Walsh, J.G., Cullen, S.P., Sheridan, C., Luthi, AU., Gerner, C., Martin, S.J. (2008) Executioner caspase-3 and caspase-7 are functionally distinct proteases. Proc Natl Acad Sci USA ;105(35):12815-9. doi: 10.1073/pnas.0707715105. Epub 2008 Aug 22. Wu, Xiaoling et al. (2014) A Caspase-Dependent Pathway Is Involved in Wnt/β-Catenin Signaling Promoted Apoptosis in Bacillus Calmette-Guerin Infected RAW264.7 Macrophages. International Journal of Molecular Sciences 15.3: 5045–5062. PMC. Web. 26 Dec. 2017. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69663 | - |
dc.description.abstract | 細胞凋亡是生物體內自發性的一種細胞程序性死亡,其參與生物體內許多重要的生理機制,像是胚胎發生、免疫反應、及神經退形性疾病等,生物體內的細胞凋亡主要是由一群專一性極高的蛋白脢-胱天蛋白酶(caspase)所調控。在渦蟲和非洲爪蟾的實驗中已經發現,caspase在再生的過程中亦扮演極為重要的角色;另一方面,經典Wnt/β-catenin訊息傳遞路徑參與發生過程中許多重要的程序,包含細胞分裂、細胞遷移、體軸建立及組織再生。研究顯示經典Wnt/β-catenin訊息傳遞路徑和胱天蛋白酶可互相調控並且影響生物體的再生。瓢體蟲(Aeolosoma viride)是一種具有極強再生能力的水生環節動物,因此在此篇論文中被用來作為模式生物進行細胞凋亡與經典Wnt/β-catenin訊息傳遞路徑調控再生的研究。
此篇論文的研究結果確認了細胞凋亡會於前端再生早期及晚期出現於再生組織中。三個與細胞凋亡相關的基因被從瓢體蟲身上選殖出來,再經由序列及演化樹的分析確認這三個基因分別為:caspase、Bcl-associated X protein (bax)、B-cell lymphoma extra large(Bcl-xL)。其中caspase跟目前已知caspase構造不盡相同,故命名為:Avi-caspase X。會促進細胞凋亡進行的Avi-caspase X 及Avi-bax均會在再生晚期大量的表現;而具有抑制細胞凋亡功能的Avi-Bcl-xL則是在再生早期大量表現並於24小時後逐漸降低。在使用RNA干擾抑制Avi-caspase X 基因表現量時,再生成功率會大幅的下降,說明了Avi-caspase X 對於瓢體蟲前端再生的重要性。這些結果均顯示細胞凋亡及caspase參與了瓢體蟲前端再生的過程。另一方面,我們發現瓢體蟲前端再生的過程中,Avi-Wnt-4的基因表現量會顯著的上升;以Wnt抑制劑 XAV939抑制Wnt訊息傳遞路徑活性時,會使再生完成率及Avi-caspase 6和新發現的Avi-caspase X的基因表現量都會顯著的下降;Bax的表現量亦會隨著caspase的表現量下降,而抑制性的Bcl-xL則會上升。但是,當Avi-caspase X 的活性被抑制時,Avi-Wnt-4的表現量並沒有產生明顯的變化。因此,推論WNT應該屬於細胞凋亡的訊息傳遞路徑的上游,可共同調控再生的進行。 | zh_TW |
dc.description.abstract | Many invertebrates possess extraordinary regenerative ability to repair their lost tissues or organs. Apoptotic caspases are required during early stage regeneration in Xenopus laevis and many model organisms. Previous study showed that Aelosoma viride could not complete its anterior regeneration when Avi-caspase 6 was knocked down by RNA interference. However, caspase was not the only factor affecting anterior regeneration in A. viride. The canonical wnt/β-Catenin signaling pathway had been prove to be involved in many biological processes such as cell proliferation, cell migration, carcinogenesis, axis formation, and tissue regeneration. Recent studies in Hydra vulgaris also indicated that wnt/β-Catenin signaling might have promoted apoptosis through a caspase-dependent pathway. In A. viride, expression of Avi-wnt-4 elevated in the anterior region during anterior regeneration, and Wnt antagonist, XAV939, showed a significant inhibitory effect during anterior regeneration.
In this study, I have identified a novel caspase, Avi-caspase X. Its expression was elevated during anterior regeneration, and was confirmed by both in situ hybridization and quantitative real-time polymerase chain reaction (qRT-PCR). In order to identify the relationship between apoptotic caspase and canonical Wnt signaling pathway, inhibitor of Wnt signaling pathway was applied to observed the change in caspase gene expression. Surprisingly, expression of both Avi-caspase X and Avi-caspase 6 decreased significantly in XAV939 treated group. The Bcl-2 protein family is notable for their regulation of apoptosis. In order to rule out the possibilities of genetic cross talk, two target genes, Bcl-2 associated X protein (apoptosis regulator Bax) and B-cell lymphoma-extra-large (Bcl-xL) from the Bcl-2 family were identified and examined. The gene expression level of Avi-Bax and Avi-Bcl-xL fluctuated during anterior regeneration, and seems to be negatively correlated to each other. Under the treatment with XAV939, the expression level of Avi-Bax decreased along with an increase in gene expression level of Avi-Bcl-xL. Finally, RNA interference (RNAi) of Avi-caspase X was performed by siRNA, further confirmed the relationship between Wnt signaling pathway and apoptotic caspase. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T03:22:54Z (GMT). No. of bitstreams: 1 ntu-107-R04b21025-1.pdf: 3573237 bytes, checksum: 6aa23e10fbbdc5f674411ce86316b5ab (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | Contents
致謝 iii 中文摘要 vii Abstract viii Introduction 1 Regeneration 1 Caspase 2 Apoptosis 3 Roles of caspase in the process of development and regeneration 4 Canonical Wnt signaling pathway 5 Aeolosoma viride 6 Material Methods 8 Aeolosoma viride 8 RNA Extraction 8 Reverse Transcription 9 Real time quantitative polymerase chain reaction (RT-qPCR) 10 in situ hybridization 10 TUNEL assay 12 dsRNA 13 Microinjection 14 Statistical analysis 14 Results 15 The number of apoptotic cells during anterior regeneration 15 Sequencing of Avi-caspase X, Avi-Bax, and Avi-Bcl-xL 15 Gene expression of Avi-caspase X, Avi-caspase 6, Avi-Bax, and Avi-Bcl-xL during anterior regeneration 18 Location of gene expression of Avi-caspases at the blastema during anterior regeneration 19 Inhibition of Avi-caspase X by dsRNA 20 Inhibitory effect of XAV939 on anterior regeneration in A. viride 21 The influence of XAV939 on apoptosis during regeneration 21 Avi-caspase X dsRNA showed no effect on the gene expression of Avi-Wnt-4 22 Discussion 24 Apoptosis and regeneration 24 Identification of a novel caspase, Avi-caspase X 24 Avi-caspase X expression is critical during anterior regeneration 25 Avi-caspase X and Avi-caspase 6 26 Relationship between apoptotic caspase and canonical Wnt signaling pathway 27 Compensatory proliferation in A. viride 28 Reference 29 Tables 35 Table 1. primers used for cloning apoptotic related genes 35 Table 2. primers used for the RNA probes 35 Table 3. primers used for qPCR 36 Table 4. primers used for the synthesis of dsRNA 36 Figures 37 Figure 1. Detection of apoptotic cells during anterior regeneration 39 Figure 2. Sequence of Avi-caspase X, Avi-Bax, Avi-Bcl-xL 46 Figure 3. Phylogenetic tree for Avi-caspases 48 Figure 4. Phylogenetic tree for Avi-Bax and Avi-Bcl-xL 50 Figure 5. Relative gene expression of Avi-caspases during anterior regeneration 52 Figure 6. Relative gene expression of apoptotic related genes during anterior regeneration 54 Figure 7. Localization of Avi-caspases during anterior regeneration 57 Figure 8. Knock-down experiment of Avi-caspase X by dsRNA 60 Figure 9. The inhibitory effect of XAV939 on anterior regeneration of A. viride. 62 Figure 10. The inhibitory effect of XAV939 on anterior regeneration of A. viride. 64 Figure 11. Gene expression of Avi-caspases, Avi-Bax, and Avi-Bcl-xL decreased under XAV939 treatment 66 Figure 12. Avi-caspase X dsRNA treatment showed no significant effect on gene expression of Avi-Wnt-4 68 Figure 13. Apoptotic signals increased after XAV939 treatment 71 | |
dc.language.iso | en | |
dc.title | Avi-caspase X在瓢體蟲前端再生中的表現量是由
經典Wnt/β-catenin訊息傳遞路徑所調控 | zh_TW |
dc.title | Avi-caspase X Expression in Anterior Regeneration of Aeolosoma viride Is Regulated by the Wnt/β-Catenin Signaling Pathway | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 郭典翰(Dian-Han Kuo),吳益群(Yi-Chun Wu),蔡素宜(Su-Yi Tsai) | |
dc.subject.keyword | 瓢體蟲,再生,細胞凋亡,胱天蛋白?,Wnt 訊息傳遞路徑, | zh_TW |
dc.subject.keyword | Aeolosoma viride,regeneration,Caspase,Wnt signaling pathway,XAV939,compensatory proliferation, | en |
dc.relation.page | 71 | |
dc.identifier.doi | 10.6342/NTU201800952 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-06-15 | |
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 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-107-1.pdf 目前未授權公開取用 | 3.49 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。