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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 許弘明(Hong-Ming Hsu) | |
| dc.contributor.author | Yen-Ju Chen | en |
| dc.contributor.author | 諶彥汝 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:18:12Z | - |
| dc.date.available | 2021-11-08 | |
| dc.date.available | 2022-11-24T03:18:12Z | - |
| dc.date.copyright | 2021-11-08 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-04 | |
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FEBS letters, 2007. 581(16): p. 2919-2928. 38. Rendón-Gandarilla, F.J., et al., The TvLEGU-1, a legumain-like cysteine proteinase, plays a key role in Trichomonas vaginalis cytoadherence. BioMed Research International, 2013. 2013. 39. Van den Bout, I. and N. Divecha, PIP5K-driven PtdIns (4, 5) P2 synthesis: regulation and cellular functions. Journal of cell science, 2009. 122(21): p. 3837-3850. 40. Mandal, K., Review of PIP2 in Cellular Signaling, Functions and Diseases. International Journal of Molecular Sciences, 2020. 21(21): p. 8342. 41. Weernink, P.A.O., M. Schmidt, and K.H. Jakobs, Regulation and cellular roles of phosphoinositide 5-kinases. European journal of pharmacology, 2004. 500(1-3): p. 87-99. 42. Van Rheenen, J., et al., EGF-induced PIP2 hydrolysis releases and activates cofilin locally in carcinoma cells. The Journal of cell biology, 2007. 179(6): p. 1247-1259. 43. Dias-Lopes, G., et al., Morphologic Study of the Effect of Iron on Pseudocyst Formation in Trichomonas Vaginalis and Its Interaction With Human Epithelial Cells. Memorias do Instituto Oswaldo Cruz, 2017. 112(10): p. 664-673. 44. 林書帆, 陰道滴蟲 PI4P5K 參與在鐵刺激 PIP2 產生的功能角色. 臺灣大學微生物學研究所學位論文, 2018: p. 1-60. 45. Tsai, C.-D., H.-W. Liu, and J.-H. Tai, Characterization of an iron-responsive promoter in the protozoan pathogen Trichomonas vaginalis. Journal of Biological Chemistry, 2002. 277(7): p. 5153-5162. 46. Ryu, J., et al., Effect of iron on the virulence of Trichomonas vaginalis. Journal of Parasitology, 2001. 87(2): p. 457-460. 47. 李由, 陰道滴蟲 Myb2 及 Myb3 轉錄因子受鐵及過氧化氫誘導入核之調控機制. 臺灣大學微生物學研究所學位論文, 2011: p. 1-112. 48. Brugerolle, G., G. Bricheux, and G. Coffe, Centrin protein and genes in Trichomonas vaginalis and close relatives. Journal of Eukaryotic Microbiology, 2000. 47(2): p. 129-138. 49. Crouch, M.-L. and J.F. Alderete, Trichomonas vaginalis interactions with fibronectin and laminin. Microbiology, 1999. 145(10): p. 2835-2843. 50. Abràmoff, M.D., P.J. Magalhães, and S.J. Ram, Image processing with ImageJ. Biophotonics international, 2004. 11(7): p. 36-42. 51. Schneider, C.A., W.S. Rasband, and K.W. Eliceiri, NIH Image to ImageJ: 25 years of image analysis. Nature methods, 2012. 9(7): p. 671-675. 52. 王凱萱, 致病性原蟲陰道滴蟲細胞中 F-actin capping protein 之功能探討. 臺灣大學微生物學研究所學位論文, 2019: p. 1-74. 53. Holz, R.W., et al., A pleckstrin homology domain specific for phosphatidylinositol 4, 5-bisphosphate (PtdIns-4, 5-P2) and fused to green fluorescent protein identifies plasma membrane PtdIns-4, 5-P2 as being important in exocytosis. Journal of Biological Chemistry, 2000. 275(23): p. 17878-17885. 54. Crouch, M.V. and J.F. Alderete, Trichomonas vaginalis has two fibronectin-like iron-regulated genes. Arch Med Res, 2001. 32(2): p. 102-7. 55. Dias-Lopes, G., et al., Morphologic study of the effect of iron on pseudocyst formation in Trichomonas vaginalis and its interaction with human epithelial cells. Memórias do Instituto Oswaldo Cruz, 2017. 112(10): p. 664-673. 56. Ribeiro, K.C., L.H. MONTEIRO‐LEAL, and M. Benchimol, Contributions of the axostyle and flagella to closed mitosis in the protists Tritrichomonas foetus and Trichomonas vaginalis. Journal of Eukaryotic Microbiology, 2000. 47(5): p. 481-492. 57. Kawai, T., et al., Polarized PtdIns (4, 5) P2 distribution mediated by a voltage-sensing phosphatase (VSP) regulates sperm motility. Proceedings of the National Academy of Sciences, 2019. 116(51): p. 26020-26028. 58. Raucher, D., et al., Phosphatidylinositol 4, 5-bisphosphate Functions as a Second Messenger That Regulates Cytoskeleton-Plasma Membrane Adhesion. Cell, 2000. 100(2): p. 221-228. 59. Brown, H.A., et al., ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity. Cell, 1993. 75(6): p. 1137-1144. 60. Hammond, S.M., et al., Human ADP-ribosylation Factor-activated Phosphatidylcholine-specific Phospholipase D Defines a New and Highly Conserved Gene Family (∗). Journal of Biological Chemistry, 1995. 270(50): p. 29640-29643. 61. Jenkins, G.H., P.L. Fisette, and R.A. Anderson, Type I phosphatidylinositol 4-phosphate 5-kinase isoforms are specifically stimulated by phosphatidic acid. Journal of Biological Chemistry, 1994. 269(15): p. 11547-11554. 62. Shibasaki, Y., et al., Massive actin polymerization induced by phosphatidylinositol-4-phosphate 5-kinase in vivo. Journal of Biological Chemistry, 1997. 272(12): p. 7578-7581. 63. Wen, P.J., et al., Phosphatidylinositol (4, 5) bisphosphate coordinates actin-mediated mobilization and translocation of secretory vesicles to the plasma membrane of chromaffin cells. Nature communications, 2011. 2(1): p. 1-11. 64. Jesus, J.B.d., et al., Trichomonas vaginalis virulence against epithelial cells and morphological variability: the comparison between a well-established strain and a fresh isolate. Parasitology Research, 2004. 93(5): p. 369-377. 65. Kuvibidila, S.R., et al., Iron deficiency reduces the hydrolysis of cell membrane phosphatidyl inositol-4, 5-bisphosphate during splenic lymphocyte activation in C57BL/6 mice. The Journal of nutrition, 1998. 128(7): p. 1077-1083. 66. Schneider-Poetsch, T., et al., Inhibition of eukaryotic translation elongation by cycloheximide and lactimidomycin. Nature chemical biology, 2010. 6(3): p. 209-217. 67. Rocha-Perugini, V., M. Gordon-Alonso, and F. Sánchez-Madrid, PIP2: choreographer of actin-adaptor proteins in the HIV-1 dance. Trends in microbiology, 2014. 22(7): p. 379-388. 68. Hairfield, M.L., C. Westwater, and J.W. Dolan, Phosphatidylinositol-4-phosphate 5-kinase activity is stimulated during temperature-induced morphogenesis in Candida albicans. Microbiology, 2002. 148(6): p. 1737-1746. 69. Xie, Y., et al., Orchestrated actin nucleation by the Candida albicans polarisome complex enables filamentous growth. Journal of Biological Chemistry, 2020. 295(44): p. 14840-14854. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80830 | - |
| dc.description.abstract | 陰道滴蟲寄生於泌尿生殖道已發展出許多機制來面對多重環境變化造成的壓力。其中，已知鐵可誘發相關訊息傳導路徑來影響其生理反應，因此在陰道滴蟲內受到嚴密調控。在前人研究中發現，鐵能活化陰道滴蟲中的TvPI4P5K 來加速PIP2生成。PIP2在高等真核細胞中作為第二信使參與多種反應，其中包含肌動蛋白重組介導之細胞形變與運動；本研究旨在探討陰道滴蟲中，PIP2與其細胞骨架重組能力的關聯性。觀察實驗室蟲株，低貼附蟲株主要以鞭毛擺動而自由泳動，高貼附蟲株具有極強外觀形變能力，細胞貼附後轉以偽足構造爬行，推測細胞骨架重組能力在兩者間應有顯著差異。根據墨點分析合併免疫螢光染色，低貼附力蟲株其PIP2表現量低，訊號集中於鞭毛；在高黏附蟲株中PIP2表現量高且分布於細胞膜周圍，但在細胞貼附後細胞膜上PIP2訊號下降，顯示PIP2的表現與分布可能與陰道滴蟲細胞形變以及運動能力有關。目前實驗室自製的抗體，可應用於免疫螢光、免疫斑點以及免疫沉澱實驗，發現高貼附性蟲株比低貼附性蟲株有較高的TvPI4P5K表現量，同時有鐵環境下的蟲體也比低鐵環境中蟲體有更高的TvPI4P5K表現。而在鐵短暫刺激下調控的TvPI4P5K與PIP2表現量升高可能透過蛋白質轉譯相關路徑調控。TvPI4P5K相關的分子機制與細胞生物功能仍待進一步研究。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:18:12Z (GMT). No. of bitstreams: 1 U0001-0110202114255200.pdf: 2853450 bytes, checksum: 742f4a7b4ea47851f6309a1126e51028 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "目錄 口試委員會審定書 i 誌謝 ii 中文摘要 iii 英文摘要 iv 第一章、前言 1 第一節、陰道滴蟲簡介 1 第二節、陰道滴蟲型態 1 第三節、陰道滴蟲致病機轉 1 第四節、鐵離子與陰道滴蟲 3 第五節、磷脂酸肌醇4, 5-雙磷酸 (Phosphatidylinositol 4,5-bisphosphate, PIP2 ) 3 第六節、研究目的 5 第二章、材料與方法 6 第一節、陰道滴蟲蟲株 6 1.1蟲株種類 6 1.2蟲株培養 6 1.3陰道滴蟲之保存與活化 6 第二節、免疫螢光染色法 (Immunofluorescence assay，IFA) 7 2.1 直接固定細胞 7 2.2 活細胞自然貼附後固定 7 2.3 免疫螢光抗體反應 7 第三節、免疫斑點分析 (Dot blot) 8 第四節、蛋白質聚丙烯醯胺凝膠電泳 (SDS polyacrylamide gel electrophoresis ) 8 第五節、西方墨點法 (Western blot assay) 8 第六節、免疫沉澱法 (Immunoprecipitation , IP) 9 第三章 、 結果 10 第一節、PIP2在陰道滴蟲蟲株的表現差異 10 第二節、PIP2在陰道滴蟲細胞貼附前後的表現差異 10 第三節、實驗室自製TvPI4P5K抗體 11 第四節、TvPI4P5K抗體的專一性測試 11 第五節、TvPI4P5K在不同陰道滴蟲分離株的表現差異 13 第六節、PIP2以及TvPI4P5K在臨床分離株的表現量 13 第七節、環境中鐵離子對陰道滴蟲PIP2與TvPI4P5K的表現量影響 14 第四章 、 討論 16 第一節、PIP2以及TvPI4P5K對於陰道滴蟲細胞貼附力之關聯 16 第二節、鐵離子對於陰道滴蟲PIP2以及TvPI4P5K表現之影響 18 第三節、鐵離子可能透過調控TvPI4P5K生成進而影響PIP2表現量 19 第四節、在已知系統PIP2與病原體致病性關聯 19 參考資料 49 附錄 55 附錄一、TYI-S33培養液 55 附錄二、各實驗使用之抗體 56 附錄三、綜合抑制劑 59 附錄四、Phosphotase Inhibitor 60 附圖目錄 圖一、PIP2於不同表現型陰道滴蟲的表現差異 21 圖二、PIP2於陰道滴蟲進行細胞貼附前後的表現量與細胞內的分布 23 圖三、anti-TvPI4P5K生產抗體之peptide區域 25 圖四、TvPI4P5K抗體於陰道滴蟲全細胞裂解蛋白的專一性測試 27 圖五、免疫沉澱濃縮的TvPI4P5K抗體訊號於基因轉殖的陰道滴蟲的表現 29 圖六、TvPI4P5K抗體的專一性測試 31 圖七、濃縮的TvPI4P5K抗體訊號於不同表現型陰道滴蟲的表現 33 圖八、TvPI4P5K於不同表現型陰道滴蟲的表現差異 35 圖九、PIP2以及TvPI4P5K於不同臨床陰道滴蟲的表現差異 37 圖十、PIP2以及TvPI4P5K在不同鐵離子環境下於陰道滴蟲的表現差異 40 圖十一、PIP2以及TvPI4P5K在短暫鐵離子刺激下於陰道滴蟲的影響 42 圖十二、蛋白質合成抑制劑Cycloheximide對於PIP2以及TvPI4P5K的影響 44 圖十三、TvPI4P5K於陰道滴蟲的作用機制圖 47" | |
| dc.language.iso | zh-TW | |
| dc.subject | 細胞貼附力 | zh_TW |
| dc.subject | 陰道滴蟲 | zh_TW |
| dc.subject | 磷酸肌醇激酶 | zh_TW |
| dc.subject | 磷脂酸肌醇4 | zh_TW |
| dc.subject | 5-雙磷酸 | zh_TW |
| dc.subject | 鐵離子 | zh_TW |
| dc.subject | 細胞變形作用 | zh_TW |
| dc.subject | Phosphatidylinositol-4-phosphate-5-kinase | en |
| dc.subject | Cytoadherence | en |
| dc.subject | Morphological transformation | en |
| dc.subject | Iron | en |
| dc.subject | PIP2 | en |
| dc.subject | Trichomonas vaginalis | en |
| dc.title | 不同陰道滴蟲分離株磷脂酰肌醇4-磷酸5-激酶表現量差異分析 | zh_TW |
| dc.title | The expression profiling of a Phosphatidylinositol-4-phosphate-5-kinase in different isolates of Trichomonas vaginalis | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蔡欣祐(Hsin-Tsai Liu),許書豪(Chih-Yang Tseng) | |
| dc.subject.keyword | 陰道滴蟲,磷酸肌醇激酶,磷脂酸肌醇4, 5-雙磷酸,鐵離子,細胞變形作用,細胞貼附力, | zh_TW |
| dc.subject.keyword | Trichomonas vaginalis,Phosphatidylinositol-4-phosphate-5-kinase,PIP2,Iron,Morphological transformation,Cytoadherence, | en |
| dc.relation.page | 60 | |
| dc.identifier.doi | 10.6342/NTU202103497 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-10-04 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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