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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李宣書(Hsuan-Shu Lee) | |
dc.contributor.author | Cheng-Han Wu | en |
dc.contributor.author | 巫承翰 | zh_TW |
dc.date.accessioned | 2021-06-16T03:06:19Z | - |
dc.date.available | 2016-08-11 | |
dc.date.copyright | 2015-08-11 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-06-25 | |
dc.identifier.citation | Agata, K., Y. Saito, and E. Nakajima. 2007. Unifying principles of regeneration I: Epimorphosis versus morphallaxis. Development, growth & differentiation. 49:73-78.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/54590 | - |
dc.description.abstract | 有尾目兩棲動物具有非凡的肢體再生能力,當墨西哥蠑螈肢體被截斷後,將會啟動傷口癒合反應,接著形成原基組織並且再生出全新的肢體,在肢體再生的起始階段,原基細胞是由神經和頂端上皮蓋所誘導產生,神經在肢體再生中的作用一直以來都被頻繁的研究,雖然目前對於蠑螈肢體再生的神經依賴性已經有一定程度的了解,但是在探討神經依賴性底層機制的過程中,大規模系統化的探索還是有其必要性;在本篇研究中,高通量定序技術被用來比對再生肢體和去神經肢體的大量轉錄體,實驗的結果揭露多種原基組織和神經依賴性相關的生物程序,在將來一定能夠助益蠑螈肢體再生相關之研究;此外,我們藉由觀察蠑螈手臂再生時肌肉末端Pax7轉錄體的表現情況,對於蠑螈從原基中期到分化晚期的持續性去分化這現象,提供了扎實的證據,並且利用活體擴散張量影像技術和免疫染色實驗觀察到新生肌肉和殘餘肌肉之間具有一個接合性縫隙,綜合來看,由以上研究結果顯示出殘餘肌肉的持續性去分化現象,使得殘餘肌肉細胞保持較未分化的狀態,將能夠幫助新、殘餘肌肉組織準確的接合;這些實驗結果也許在不遠的將來,能夠幫助了解肢體再生的機制和協助再生醫學的發展。 | zh_TW |
dc.description.abstract | Urodele amphibians have an extraordinary ability to regenerate lost limbs. Amputation of the limb of axolotl causes an initial wound healing response followed by blastema formation and the regeneration of the new limb. In initial stage of limb regeneration, blastemal cells are induced by nerves and the apical epithelial cap. Nerve functions have long been the focus of limb regeneration studies. Although considerable progress has been made in inquiring into the nerve-dependency of salamander, a broader systemic perspective is still needed to investigate the underlying mechanisms upon the presence of a nerve. In this study, a high-throughput sequencing approach was used to compare transcript abundance among regenerating limbs, and limbs denervated at the time of amputation. This result addressed the biological processes which are associated with blastemal formation and nerve-dependency, and will greatly assist future research of limb regeneration in axolotl. Furthermore, I added evidence that dedifferentiation persisted from mid-bud to late differentiation stage by showing expression of Pax7 transcripts in the remnant muscle fiber ends. Moreover, in vivo diffusion tensor imaging and stained tissue sections indicate a gap between the regenerating and parental muscles. Taken together, the present data suggested that prolonged dedifferentiation at remnant muscle ends might facilitate accurate connection between regenerating and parental muscle fibers due to their immature status. Additionally, these results may assist in the understanding of limb regeneration, ultimately facilitating the development of regenerative medicine in mammals. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T03:06:19Z (GMT). No. of bitstreams: 1 ntu-104-D97642006-1.pdf: 4904674 bytes, checksum: 8ef44b4a165ae7b132351230e92c8386 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 口試委員會審定書 I
致謝 III 中文摘要 IV ABSTRACT V CONTENTS VII LIST OF FIGURES XI LIST OF TABLES XIV LIST OF ABBREVIATIONS XV CHAPTER 1: LITERATURE REVIEW 1 1.1 Introduction 2 1.2 Overview of Regeneration 4 1.2.1 Cellular and molecular fundamentals 4 1.2.2 Regeneration in animals 7 1.3 Salamander and Regeneration 12 1.3.1 Salamander 12 1.3.2 Use as a model organism 14 1.3.3 Origin of Blastema 16 1.4 Hypothesis and aims 18 CHAPTER 2: DE NOVO TRANSCRIPTOME SEQUENCING OF AXOLOTL BLASTEMA FOR IDENTIFICATION OF DIFFERENTIALLY EXPRESSED GENES DURING LIMB REGENERATION 21 2.1 Summary 22 2.2 Introduction 24 2.3 Materials and Methods 28 2.3.1 Animal experimental procedures 28 2.3.2 RNA extraction and library preparation 28 2.3.3 Analysis of Illumina sequencing results 29 2.3.4 Identification of differentially expressed genes 31 2.3.5 qPCR for mRNA quantification 32 2.3.6 In situ hybridization 32 2.4 Results 35 2.4.1 Illumina NGS and read assembly 35 2.4.2 Annotation of predicted proteins 36 2.4.3 Gene ontology and classification of clusters of orthologous groups 37 2.4.4 Detection of growth factor and transcription factor sequences related to limb regeneration in axolotls 38 2.4.5 Comparing gene expression profiles between blastema and denervated limb stump 40 2.4.6 Functional annotation of differentially expressed genes 42 2.4.7 Validation of differentially expressed genes by qPCR 42 2.4.8 Expression patterns of differentially expressed genes in limb regeneration 43 2.4.9 MS-275 inhibits limb regeneration in A. mexicanum larvae 44 2.4.10 HDACs are required for limb regeneration in A. mexicanum larvae 46 2.5 Discussion 48 2.6 Tables and Figures 50 CHAPTER 3: LONG-DURATION MUSCLE DEDIFFERENTIATION DURING LIMB REGENERATION IN AXOLOTLS 67 3.1 Summary 68 3.2 Introduction 69 3.3 Materials and Methods 72 3.3.1 Animal experimental procedures 72 3.3.2 Sequencing of axolotl Pax7 mRNA 72 3.3.3 Cloning of the coding sequence of axolotl Pax7 cDNA and preparation of RNA probes 74 3.3.4 Whole-mount in situ hybridization of Pax7 transcripts in axolotl embryos 75 3.3.5 Preparation of tissue sections 76 3.3.6 Tissue section in situ hybridization 76 3.3.7 Immunohistochemistry 77 3.3.8 Microscopy and image processing 78 3.4 Results 79 3.4.1 Nucleotide and deduced amino acid sequence of axolotl Pax7 79 3.4.2 Expression of Pax7 mRNA in axolotl embryos 80 3.4.3 Appearance of Pax7 mRNA expression in the muscle stump during the early bud and mid-bud stages 80 3.4.4 Immunohistochemical staining of muscle ends in longitudinal sections: mid-bud stage 81 3.4.5 Immunohistochemical staining of muscle ends in transverse sections: early bud and mid-bud stages 83 3.4.6 Lack of proliferation of PAX7+ cells in distal muscle ends at the mid-bud stage 84 3.4.7 Muscle regeneration during the early differentiation stage 85 3.4.8 Muscle regeneration during the late differentiation stage 86 3.4.9 Expression of Pax7 transcripts in the remnant muscle ends during early and late differentiation stages 87 3.5 Discussion 87 3.6 Figures 95 CHAPTER 4: DEMONSTRATION OF CONNECTION BETWEEN REGENERATING AND PARENTAL MUSCLES DURING AXOLOTL LIMB REGENERATION BY DIFFUSION TENSOR IMAGING 108 4.1 Summary 109 4.3 Materials and Methods 114 4.3.1 Animal experimental procedures 114 4.3.2 Preparation of tissue sections 114 4.3.3 Immunohistochemistry 115 4.3.4 Microscopy and image processing 116 4.3.5 DTI and MRI of axolotl muscle fibers 116 4.4.1 Muscle fiber tractography of a right upper arm 119 4.4.2 Fiber tractography of muscles during regeneration 120 4.4.3 Connection between regenerating and parental muscle fibers observed by DTI 121 4.5 Discussion 122 4.6 Figures 125 PROSPECTIVE ASPECTS 132 APPENDIX 141 REFERENCES 145 LIST OF PUBLICATIONS 176 | |
dc.language.iso | en | |
dc.title | 於蠑螈手臂再生時尋找其差異性表達基因和證明持續性肌肉去分化 | zh_TW |
dc.title | Identification of differentially expressed genes and demonstration of long-duration muscle dedifferentiation during limb regeneration in axolotls | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 宋麗英(Li-Ying Sung),林劭品(Sau-Ping Lin),李士傑(Shyh-Jye Lee),曾文毅(Wen-Yih Isaac Tseng),謝清河(Patrick C.H. Hsieh) | |
dc.subject.keyword | 原基,肢體再生,神經依賴性,墨西哥蠑螈,去分化,肌肉,擴散張量影像,再生醫學, | zh_TW |
dc.subject.keyword | blastemal,limb regeneration,nerve-dependency,axolotl,dedifferentiation,muscle,diffusion tensor imaging,regenerative medicine, | en |
dc.relation.page | 176 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-06-25 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 生物科技研究所 | zh_TW |
顯示於系所單位: | 生物科技研究所 |
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