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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46131完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李士傑 | |
| dc.contributor.author | Hsin-Ting Wu | en |
| dc.contributor.author | 吳欣庭 | zh_TW |
| dc.date.accessioned | 2021-06-15T04:54:58Z | - |
| dc.date.available | 2012-09-21 | |
| dc.date.copyright | 2011-09-21 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-18 | |
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Human homologue of mouse lymph node homing receptor: evolutionary conservation at tandem cell interaction domains. Proc Natl Acad Sci U S A 86:5562-5566. Strubel NA, Nguyen M, Kansas GS, Tedder TF, Bischoff J. 1993. Isolation and characterization of a bovine cDNA encoding a functional homolog of human P-selectin. Biochem Biophys Res Commun 192:338-344. Sun G, Pan J, Liu K, Wang X, Wang S. 2010. Molecular Cloning and Expression Analysis of P-Selectin from Zebrafish (Danio rerio). Int J Mol Sci 11:4618-4630. Tedder TF, Isaacs CM, Ernst TJ, Demetri GD, Adler DA, Disteche CM. 1989. Isolation and chromosomal localization of cDNAs encoding a novel human lymphocyte cell surface molecule, LAM-1. Homology with the mouse lymphocyte homing receptor and other human adhesion proteins. J Exp Med 170:123-133. Tedder TF, Steeber DA, Pizcueta P. 1995. L-selectin-deficient mice have impaired leukocyte recruitment into inflammatory sites. J Exp Med 181:2259-2264. Tsao MC. 2009. Knockdown of LPA1 inhibits neutrophil recruitment during wound healing. Master Thesis. Tsurushita N, Fu H, Berg EL. 1996. PCR cloning of the cDNA encoding baboon L-selectin. Gene 181:219-220. Vestweber D, Blanks JE. 1999. Mechanisms that regulate the function of the selectins and their ligands. Physiol Rev 79:181-213. von Andrian UH, Chambers JD, Berg EL, Michie SA, Brown DA, Karolak D, Ramezani L, Berger EM, Arfors KE, Butcher EC. 1993. L-selectin mediates neutrophil rolling in inflamed venules through sialyl LewisX-dependent and -independent recognition pathways. Blood 82:182-191. von Andrian UH, Chambers JD, McEvoy LM, Bargatze RF, Arfors KE, Butcher EC. 1991. Two-step model of leukocyte-endothelial cell interaction in inflammation: distinct roles for LECAM-1 and the leukocyte beta 2 integrins in vivo. Proc Natl Acad Sci U S A 88:7538-7542. Weller A, Isenmann S, Vestweber D. 1992. Cloning of the mouse endothelial selectins. Expression of both E- and P-selectin is inducible by tumor necrosis factor alpha. J Biol Chem 267:15176-15183. Yan HC, Delisser HM, Pilewski JM, Barone KM, Szklut PJ, Chang XJ, Ahern TJ, Langer-Safer P, Albelda SM. 1994. Leukocyte recruitment into human skin transplanted onto severe combined immunodeficient mice induced by TNF-alpha is dependent on E-selectin. J Immunol 152:3053-3063. Zarbock A, Polanowska-Grabowska RK, Ley K. 2007. Platelet-neutrophil-interactions: linking hemostasis and inflammation. Blood Rev 21:99-111. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46131 | - |
| dc.description.abstract | 選擇蛋白(selectin)是一種黏著分子，在哺乳類的研究中顯示其可調節白血球滾動、復位和再循環。選擇蛋白有三種，分別是：L-selectin以游離形式出現在血清中，也以跨膜蛋白的形式出現在嗜中性白血球、單核球、Ｔ細胞、Ｂ細胞、自然殺手細胞及未成熟的造血細胞膜上；P-selectin表現在被活化的血小板及內皮細胞膜上；E-selectin表現在內皮細胞膜上。斑馬魚在最近被用來研究白血球在傷口的聚集作用，然而我們對斑馬魚選擇蛋白的所知有限。因此本研究選殖出斑馬魚的選擇蛋白，並嘗試釐清其可能的功能與表現。我選殖了三種選擇蛋白，並利用整體原位雜交方法觀察選擇蛋白在斑馬魚的表現位置。結果顯示這三種選擇蛋白的表現位置有部分重疊。我們亦利用morpholino削弱選擇蛋白在魚體中的表現，於受精後三天將魚尾切除以製造傷口，利用螢光顯微鏡在切除魚尾8.5小時後觀察其嗜中性球的遷移情形。我發現削弱L及E選擇蛋白的表現會抑制嗜中性球遷移到傷口處，削弱Ｐ選擇蛋白的表現則會在製造傷口後前兩小時促進嗜中性球的遷移。因此我們推測斑馬魚的選擇蛋白在調節白血球於傷口的聚集作用上，有與哺乳類選擇蛋白類似的功能。 | zh_TW |
| dc.description.abstract | Selectins are adhesion molecules known to mediate leukocyte rolling, homing and recirculation. There are three types of selectins: L-selectin is in a soluble form in serum and a transmembrane protein on the membrane of neutrophils, monocytes, T and B cells, natural killer cells and immature hematopoietic cells; P-selectin is expressed in activated platelets and endothelial cells; E-selectin is expressed on endothelium cell membranes. Zebrafish is a newly established model for leukocyte recruitment study, but we know very little about selectins in zebrafish. Therefore, I aimed to investigate the roles of selectins in zebrafish. I cloned three selectins and studied their expression patterns by RT-PCR and whole-mount in situ hybridization (WISH). Zebrafish selectins share sequence homology to their respective homologs. WISH analysis showed overlapped expression patterns among three types of selectins. To characterize their roles in neutrophil recruitment, I cut the tails of Tg(MPO::EGFP)114 zebrafish at 3 days post-fertilization (dpf) and observed neutrophil recruitments at 8.5 hours post-injury (hpi) by epifluorescent microscope. Knockdown of L– or E-selectin inhibits neutrophil recruitment whereas knockdown of P-selectin enhances neutrophil recruitment at first 2 hours after injury. It suggests that zebrafish selectins may play a conserved role in mediating neutrophil recruitment in zebrafish. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T04:54:58Z (GMT). No. of bitstreams: 1 ntu-100-R98B41009-1.pdf: 1448891 bytes, checksum: 7061c6d85b15e58ce6a9f590383be625 (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 謝誌 i
中文摘要 ii Abstract iv Introduction 3 Materials and methods 7 Maintenance of Zebrafish 7 Cloning and analysis of zebrafish E-, L-, P-selectin 7 Whole-mount in situ hybridization 8 Microinjection 9 Morpholino Oligonucleotides (MO) knockdown 9 Neutrophil recruitment detection 9 Statistical analysis 10 Results 11 Sequencing analysis of zebrafish selectins 11 Expression patterns of zebrafish selectins 12 Knockdown of zebrafish selectins affects neutrophil recruitment in different degrees 14 Discussion 18 Selectin sequence analysis 18 Expression patterns 19 Selectin in neutrophil recruitment 21 References 24 Table 32 Table 1 Primer for cloning and RT-PCR 32 Table 2 Amino acid sequences similarity table of L-selectin homologues. 33 Table 3 Amino acid sequences similarity table of P-selectin homologues. 33 Table 4 Amino acid sequences similarity table of E-selectin homologues. 33 Figures 34 Fig. 1 Sequence analysis of zebrafish selectin 36 Fig. 2 Temporal and spatial expressions of selectins by RT-PCR 37 Fig. 3 Whole-mount in situ hybridization analysis of E-selectin expression patterns 38 Fig. 4 Whole-mount in situ hybridization analysis of L-selectin expression patterns 39 Fig. 5 Whole-mount in situ hybridization analysis of P-selectin expression patterns 40 Fig. 6 Knockdown of E-selectin inhibits neutrophil recruitment 42 Fig. 7 Knockdown of L-selectin inhibits neutrophil recruitment. 44 Fig. 8 Knockdown of P-selectin enhances neutrophil recruitment at the first 2 hpi but had no differences at 8.5 hpi. 46 Appendix 47 Figures 47 Fig. 1 The methods of neutrophil recruitment detection 47 Fig. 2 The nucleotide sequence of zebrafish E-selectin 49 Fig. 3 The nucleotide sequence of zebrafish partial L-selectin 50 Fig. 4 The nucleotide sequence of zebrafish P-selectin 52 | |
| dc.language.iso | zh-TW | |
| dc.subject | 發炎反應 | zh_TW |
| dc.subject | Selectin | zh_TW |
| dc.subject | 嗜中性球 | zh_TW |
| dc.subject | Recruitment | zh_TW |
| dc.subject | 斑馬魚 | zh_TW |
| dc.subject | neutrophil | en |
| dc.subject | inflammation | en |
| dc.subject | zebrafish | en |
| dc.subject | recruitment | en |
| dc.subject | selectin | en |
| dc.title | 斑馬魚selectins的選殖、表現與功能分析 | zh_TW |
| dc.title | Cloning, expression and functional analysis of zebrafish selectins | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 丁詩同,沈湯龍 | |
| dc.subject.keyword | Selectin,嗜中性球,Recruitment,斑馬魚,發炎反應, | zh_TW |
| dc.subject.keyword | selectin,neutrophil,recruitment,zebrafish,inflammation, | en |
| dc.relation.page | 52 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-18 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
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