跳到主要內容

臺灣博碩士論文加值系統

(3.239.4.127) 您好!臺灣時間:2022/08/20 06:44
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:王嘉佩
研究生(外文):Jia-Pey Wang
論文名稱:斑馬魚促細胞凋亡基因zfbax1誘發之早期胚胎細胞凋亡
論文名稱(外文):Earlier embryonic apoptosis induced by zebrafish pro-apoptotic gene zfbax1
指導教授:吳金洌吳金洌引用關係
指導教授(外文):Jen-Leih Wu
學位類別:碩士
校院名稱:國立海洋大學
系所名稱:水產生物技術研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:53
中文關鍵詞:斑馬魚細胞凋亡
外文關鍵詞:zebrafishbaxapoptosisembryo
相關次數:
  • 被引用被引用:0
  • 點閱點閱:288
  • 評分評分:
  • 下載下載:81
  • 收藏至我的研究室書目清單書目收藏:0
Bcl-2家族蛋白在動物的胚胎發育過程中具有重要的功能,然而其分子機制之進行則尚未明瞭,本實驗室以斑馬魚做為模式動物,研究Bcl-2家族蛋白在胚胎發育中細胞凋亡的功能,因此選殖出zfbax1,其屬於Bcl-2家族中具促細胞凋亡能力者,cDNA全長有1938個核苷酸,轉譯出192胺基酸,根據胺基酸序列比對結果顯示zfBax1與人類Baxα等形(isoform)具有54﹪identity,且包含三個與促細胞凋亡之Bcl-2家族蛋白高保守度之BH區域,自N端到C端依序為BH3、BH1與BH2; zfBax1在跨膜區域具有多個胺機酸與哺乳類不同,尤其哺乳類具有決定Bax在細胞中位置的186Serine,zfBax1則為186Alanine;因此將zfBax1與綠螢光蛋白作成融合蛋白(fusion protein),以追蹤zfBax1在細胞中的位置。zfBax1於細胞中表現後直接進入粒腺體,並導致細胞死亡,此異於哺乳類之Bax表現在細胞質且不會造成細胞死亡。以北方墨點法偵測斑馬魚整個胚胎發育過程中zfbax1 mRNA均有表現;另在胚胎表現EGFP-t-zfBxa1會使胚胎在blastula後期至gastrula前期進行細胞凋亡,且具有58﹪致死率,然而此現象可經由大量表現EGFP-zfMcl-1a而延緩;綜合以上結果,zfBax1歸屬於促細胞凋亡之Bcl-2家族成員,且可能在斑馬魚胚胎發育過程中具有重要的功能。

Proteins of the Bcl2 family play important roles in animal development; however, the underlying molecular mechanisms of their actions are poorly understood. As part of our undergoing effort to develop zebrafish embryos as a model system for the embryonic study of the Bcl2 family proteins, here we described the cloning and characterization of the third members of the zebrafish Bcl2 protein family, zfBax1. The zfBax1 cDNA is 1938 nucleotides long, encoding a polypeptide of 192 amino acids. The primary sequence of zfBax1 shares 54 % identity to human Baxisoform, and contains all three conserved BH domains of the pro-apoptotic Bcl-2 family proteins. Northern blot analysis detected a single mRNA species with decreasing abundance over time in oocytes and all selected stages of developing embryos. GFP-tagging analysis demonstrated a punctate cytoplasmic distribution pattern, which was overlapping with mitochondria-specific staining for the tagged zfBax1 protein in transfected cells. Over-expression of GFP-zfBax1 protein resulted in rapid and drastic cell death in transfected cells even before the expression of the fusion protein could be detected by epi-fluorescent microscopic examination. Finally, GFP-zfBax1 over-expression led to apoptotic death in embryonic cells as apparently by nuclear condensation and DNA fragmentation, and its killing effect was retarded by simultaneous over-expression of GFP-zfmcl1a protein. Overall, our results suggest that zfBax1 is a pro-apoptotic Bcl-2 family protein, and is likely to play an important role in zebrafish oogenesis and embryogenesis.

目錄
中文摘要---------------------------1
英文摘要---------------------------2
序論-----------------------------3
材料 ---------------------------9
方法-----------------------------14
結果-----------------------------21
討論---------------------------28
參考文獻--------------------------34
圖表-------------------------------39

Adams, J. M., and Cory, S. (1998). The Bcl-2 protein family: arbiters of cell survival. Science 281(5381), 1322-6.
Aladjem, M. I., Spike, B. T., Rodewald, L. W., Hope, T. J., Klemm, M., Jaenisch, R., and Wahl, G. M. (1998). ES cells do not activate p53-dependent stress responses and undergo p53-independent apoptosis in response to DNA damage. Curr Biol 8(3), 145-55.
Anderson, J. A., Lewellyn, A. L., and Maller, J. L. (1997). Ionizing radiation induces apoptosis and elevates cyclin A1-Cdk2 activity before but not after the midblastula transition in Xenopus. Mol Biol Cell 8(7), 1195-206.
Ashkenazi, A., and Dixit, V. M. (1998). Death receptors: signaling and modulation. Science 281(5381), 1305-8.
Brison, D. R., and Schultz, R. M. (1997). Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including transforming growth factor alpha. Biol Reprod 56(5), 1088-96.
Chao, D. T., and Korsmeyer, S. J. (1998). BCL-2 family: regulators of cell death. Annu Rev Immunol 16, 395-419.
Chen, M. C., Gong, H. Y., Cheng, C. Y., Wang, J. P., Hong, J. R., and Wu, J. L. (2000). Cloning and characterization of a novel nuclear Bcl-2 family protein, zfMcl-1a, in zebrafish embryo. Biochem Biophys Res Commun 279(2), 725-31.
Chen, M. C., Gong, H. Y., Cheng, C. Y., Wang, J. P., Hong, J. R., and Wu, J. L. (2001). Cloning and characterization of zfBLP1, a Bcl-XL homologue from the zebrafish, Danio rerio. Biochim Biophys Acta 1519(1-2), 127-33.
Chou, J. J., Li, H., Salvesen, G. S., Yuan, J., and Wagner, G. (1999). Solution structure of BID, an intracellular amplifier of apoptotic signaling. Cell 96(5), 615-24.
Clarke, P. G., and Clarke, S. (1996). Nineteenth century research on naturally occurring cell death and related phenomena. Anat Embryol (Berl) 193(2), 81-99.
Deng, C., Zhang, P., Harper, J. W., Elledge, S. J., and Leder, P. (1995). Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 82(4), 675-84.
Desagher, S., and Martinou, J. C. (2000). Mitochondria as the central control point of apoptosis. Trends Cell Biol 10(9), 369-77.
Ellis, H. M., and Horvitz, H. R. (1986). Genetic control of programmed cell death in the nematode C. elegans. Cell 44(6), 817-29.
Finkielstein, C. V., Lewellyn, A. L., and Maller, J. L. (2001). The midblastula transition in Xenopus embryos activates multiple pathways to prevent apoptosis in response to DNA damage. Proc Natl Acad Sci U S A 98(3), 1006-11.
Green, D. R., and Reed, J. C. (1998). Mitochondria and apoptosis. Science 281(5381), 1309-12.
Hengartner, M. O., and Horvitz, H. R. (1994). C. elegans cell survival gene ced-9 encodes a functional homolog of the mammalian proto-oncogene bcl-2. Cell 76(4), 665-76.
Hensey, C., and Gautier, J. (1997). A developmental timer that regulates apoptosis at the onset of gastrulation. Mech Dev 69(1-2), 183-95.
Herzog, K. H., Chong, M. J., Kapsetaki, M., Morgan, J. I., and McKinnon, P. J. (1998). Requirement for Atm in ionizing radiation-induced cell death in the developing central nervous system. Science 280(5366), 1089-91.
Hsu, S. Y., and Hsueh, A. J. (2000). Tissue-specific Bcl-2 protein partners in apoptosis: An ovarian paradigm. Physiol Rev 80(2), 593-614.
Ikegami, R., Hunter, P., and Yager, T. D. (1999). Developmental activation of the capability to undergo checkpoint-induced apoptosis in the early zebrafish embryo. Dev Biol 209(2), 409-33.
Inohara, N., and Nunez, G. (2000). Genes with homology to mammalian apoptosis regulators identified in zebrafish. Cell Death Differ 7(5), 509-10.
Jacobson, M. D., Weil, M., and Raff, M. C. (1997). Programmed cell death in animal development. Cell 88(3), 347-54.
Jurisicova, A., Varmuza, S., and Casper, R. F. (1996). Programmed cell death and human embryo fragmentation. Mol Hum Reprod 2(2), 93-8.
Kerr, J. F., Wyllie, A. H., and Currie, A. R. (1972). Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26(4), 239-57.
Lindsten, T., Ross, A. J., King, A., Zong, W. X., Rathmell, J. C., Shiels, H. A., Ulrich, E., Waymire, K. G., Mahar, P., Frauwirth, K., Chen, Y., Wei, M., Eng, V. M., Adelman, D. M., Simon, M. C., Ma, A., Golden, J. A., Evan, G., Korsmeyer, S. J., MacGregor, G. R., and Thompson, C. B. (2000). The combined functions of proapoptotic Bcl-2 family members bak and bax are essential for normal development of multiple tissues. Mol Cell 6(6), 1389-99.
Lockshin, R. A. (1969). Programmed cell death. Activation of lysis by a mechanism involving the synthesis of protein. J Insect Physiol 15(9), 1505-16.
Makarova, O., Kamberov, E., and Margolis, B. (2000). Generation of deletion and point mutations with one primer in a single cloning step. Biotechniques 29(5), 970-2.
McDonnell, J. M., Fushman, D., Milliman, C. L., Korsmeyer, S. J., and Cowburn, D. (1999). Solution structure of the proapoptotic molecule BID: a structural basis for apoptotic agonists and antagonists. Cell 96(5), 625-34.
Nechushtan, A., Smith, C. L., Hsu, Y. T., and Youle, R. J. (1999). Conformation of the Bax C-terminus regulates subcellular location and cell death. Embo J 18(9), 2330-41.
Norimura, T., Nomoto, S., Katsuki, M., Gondo, Y., and Kondo, S. (1996). p53-dependent apoptosis suppresses radiation-induced teratogenesis. Nat Med 2(5), 577-80.
Oltvai, Z. N., Milliman, C. L., and Korsmeyer, S. J. (1993). Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74(4), 609-19.
Puthalakath, H., Huang, D. C., O'Reilly, L. A., King, S. M., and Strasser, A. (1999). The proapoptotic activity of the Bcl-2 family member Bim is regulated by interaction with the dynein motor complex. Mol Cell 3(3), 287-96.
Reed, J. C., Miyashita, T., Takayama, S., Wang, H. G., Sato, T., Krajewski, S., Aime-Sempe, C., Bodrug, S., Kitada, S., and Hanada, M. (1996). BCL-2 family proteins: regulators of cell death involved in the pathogenesis of cancer and resistance to therapy. J Cell Biochem 60(1), 23-32.
Sattler, M., Liang, H., Nettesheim, D., Meadows, R. P., Harlan, J. E., Eberstadt, M., Yoon, H. S., Shuker, S. B., Chang, B. S., Minn, A. J., Thompson, C. B., and Fesik, S. W. (1997). Structure of Bcl-xL-Bak peptide complex: recognition between regulators of apoptosis. Science 275(5302), 983-6.
Saunders, J. W., Jr. (1966). Death in embryonic systems. Science 154(749), 604-12.
Shimizu, S., Ide, T., Yanagida, T., and Tsujimoto, Y. (2000a). Electrophysiological study of a novel large pore formed by Bax and the voltage-dependent anion channel that is permeable to cytochrome c. J Biol Chem 275(16), 12321-5.
Shimizu, S., Konishi, A., Kodama, T., and Tsujimoto, Y. (2000b). BH4 domain of antiapoptotic Bcl-2 family members closes voltage-dependent anion channel and inhibits apoptotic mitochondrial changes and cell death. Proc Natl Acad Sci U S A 97(7), 3100-5.
Shimizu, S., Narita, M., and Tsujimoto, Y. (1999). Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399(6735), 483-7.
Shimizu, S., and Tsujimoto, Y. (2000). Proapoptotic BH3-only Bcl-2 family members induce cytochrome c release, but not mitochondrial membrane potential loss, and do not directly modulate voltage-dependent anion channel activity. Proc Natl Acad Sci U S A 97(2), 577-82.
Sible, J. C., Anderson, J. A., Lewellyn, A. L., and Maller, J. L. (1997). Zygotic transcription is required to block a maternal program of apoptosis in Xenopus embryos. Dev Biol 189(2), 335-46.
Stack, J. H., and Newport, J. W. (1997). Developmentally regulated activation of apoptosis early in Xenopus gastrulation results in cyclin A degradation during interphase of the cell cycle. Development 124(16), 3185-95.
Suzuki, M., Youle, R. J., and Tjandra, N. (2000). Structure of Bax: coregulation of dimer formation and intracellular localization. Cell 103(4), 645-54.
Tata, J. R. (1966). Requirement for RNA and protein synthesis for induced regression of the tadpole tail in organ culture. Dev Biol 13(1), 77-94.
Thornberry, N. A., and Lazebnik, Y. (1998). Caspases: enemies within. Science 281(5381), 1312-6.
von Ahsen, O., Renken, C., Perkins, G., Kluck, R. M., Bossy-Wetzel, E., and Newmeyer, D. D. (2000). Preservation of mitochondrial structure and function after Bid- or Bax-mediated cytochrome c release. J Cell Biol 150(5), 1027-36.
Yang, E., Zha, J., Jockel, J., Boise, L. H., Thompson, C. B., and Korsmeyer, S. J. (1995). Bad, a heterodimeric partner for Bcl-XL and Bcl-2, displaces Bax and promotes cell death. Cell 80(2), 285-91.
Yuan, J., Shaham, S., Ledoux, S., Ellis, H. M., and Horvitz, H. R. (1993). The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1 beta-converting enzyme. Cell 75(4), 641-52.

QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊