請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45442
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 吳益群 | |
dc.contributor.author | Ya-Chi Chan | en |
dc.contributor.author | 詹雅琦 | zh_TW |
dc.date.accessioned | 2021-06-15T04:20:24Z | - |
dc.date.available | 2016-09-14 | |
dc.date.copyright | 2011-09-14 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-17 | |
dc.identifier.citation | Aizenman, E., Stout, A.K., Hartnett, K.A., Dineley, K.E., McLaughlin, B., and Reynolds, I.J. (2000). Induction of neuronal apoptosis by thiol oxidation: putative role of intracellular zinc release. J Neurochem 75, 1878-1888.
Brenner, S. (1974). The genetics of Caenorhabditis elegans. Genetics 77, 71-94. Cain, K., Langlais, C., Sun, X.M., Brown, D.G., and Cohen, G.M. (2001). Physiological concentrations of K+ inhibit cytochrome c-dependent formation of the apoptosome. J Biol Chem 276, 41985-41990. Chen, F., Hersh, B.M., Conradt, B., Zhou, Z., Riemer, D., Gruenbaum, Y., and Horvitz, H.R. (2000). Translocation of C. elegans CED-4 to nuclear membranes during programmed cell death. Science 287, 1485-1489. Chew, S.K., Chen, P., Link, N., Galindo, K.A., Pogue, K., and Abrams, J.M. (2009). Genome-wide silencing in Drosophila captures conserved apoptotic effectors. nature 460, 123-127. Conradt, B., and Horvitz, H.R. (1998). The C-elegans protein EGL-1 is required for programmed cell death and interacts with the Bcl-2-like protein CED-9. Cell 93, 519-529. Desai, C., Garriga, G., Mcintire, S.L., and Horvitz, H.R. (1988). A Genetic Pathway for the Development of the Caenorhabditis-Elegans Hsn Motor Neurons. Nature 336, 638-646. Ellis, H.M., and Horvitz, H.R. (1986). Genetic-Control of Programmed Cell-Death in the Nematode C-Elegans. Cell 44, 817-829. Ellis, R.E., Jacobson, D.M., and Horvitz, H.R. (1991). Genes Required for the Engulfment of Cell Corpses during Programmed Cell-Death in Caenorhabditis-Elegans. Genetics 129, 79-94. Fransson, A., Ruusala, A., and Aspenstrom, P. (2003). Atypical Rho GTPases have roles in mitochondrial homeostasis and apoptosis. J Biol Chem 278, 6495-6502. Galvin, B.D., Denning, D.P., and Horvitz, H.R. (2011). SPK-1, an SR protein kinase, inhibits programmed cell death in Caenorhabditis elegans. Proc Natl Acad Sci U S A 108, 1998-2003. Garland, J.M., and Halestrap, A. (1997). Energy metabolism during apoptosis. Bcl-2 promotes survival in hematopoietic cells induced to apoptose by growth factor withdrawal by stabilizing a form of metabolic arrest. J Biol Chem 272, 4680-4688. Glucksmann, A. (1951). Cell Deaths in Normal Vertebrate Ontogeny. Biol Rev 26, 59-86. Gottlob, K., Majewski, N., Kennedy, S., Kandel, E., Robey, R.B., and Hay, N. (2001). Inhibition of early apoptotic events by Akt/PKB is dependent on the first committed step of glycolysis and mitochondrial hexokinase. Genes Dev 15, 1406-1418. Hedgecock, E.M., Sulston, J.E., and Thomson, J.N. (1983). Mutations Affecting Programmed Cell Deaths in the Nematode Caenorhabditis Elegans. Science 220, 1277-1279. Hengartner, M.O., Ellis, R.E., and Horvitz, H.R. (1992). Caenorhabditis-Elegans Gene Ced-9 Protects Cells from Programmed Cell-Death. Nature 356, 494-499. Hoeppner, D.J., Hengartner, M.O., and Schnabel, R. (2001). Engulfment genes cooperate with ced-3 to promote cell death in Caenorhabditis elegans. Nature 412, 202-206. Hughes, F.M., Jr., Bortner, C.D., Purdy, G.D., and Cidlowski, J.A. (1997). Intracellular K+ suppresses the activation of apoptosis in lymphocytes. J Biol Chem 272, 30567-30576. Kerr, J.F.R., Wyllie, A.H., and Currie, A.R. (1972). Apoptosis - Basic Biological Phenomenon with Wide-Ranging Implications in Tissue Kinetics. Brit J Cancer 26, 239-&. Kimble, J., and Hirsh, D. (1979). Post-Embryonic Cell Lineages of the Hermaphrodite and Male Gonads in Caenorhabditis-Elegans. Dev Biol 70, 396-417. Knowles, L.M., Yang, C., Osterman, A., and Smith, J.W. (2008). Inhibition of fatty-acid synthase induces caspase-8-mediated tumor cell apoptosis by up-regulating DDIT4. J Biol Chem 283, 31378-31384. Koppen, M., Simske, J.S., Sims, P.A., Firestein, B.L., Hall, D.H., Radice, A.D., Rongo, C., and Hardin, J.D. (2001). Cooperative regulation of AJM-1 controls junctional integrity in Caenorhabditis elegans epithelia. Nat Cell Biol 3, 983-991. Lee, J.Y., Cole, T.B., Palmiter, R.D., and Koh, J.Y. (2000). Accumulation of zinc in degenerating hippocampal neurons of ZnT3-null mice after seizures: evidence against synaptic vesicle origin. J Neurosci 20, RC79. Neukomm, L.J., Frei, A.P., Cabello, J., Kinchen, J.M., Zaidel-Bar, R., Ma, Z., Haney, L.B., Hardin, J., Ravichandran, K.S., Moreno, S., et al. (2011). Loss of the RhoGAP SRGP-1 promotes the clearance of dead and injured cells in Caenorhabditis elegans. Nat Cell Biol 13, 79-86. Peden, E., Kimberly, E., Gengyo-Ando, K., Mitani, S., and Xue, D. (2007). Control of sex-specific apoptosis in C. elegans by the BarH homeodomain protein CEH-30 and the transcriptional repressor UNC-37/Groucho. Genes Dev 21, 3195-3207. Reddien, P.W., Andersen, E.C., Huang, M.C., and Horvitz, H.R. (2007). DPL-1 DP, LIN-35 Rb and EFL-1 E2F act with the MCD-1 zinc-finger protein to promote programmed cell death in Caenorhabditis elegans. Genetics 175, 1719-1733. Reddien, P.W., Cameron, S., and Horvitz, H.R. (2001). Phagocytosis promotes programmed cell death in C. elegans. Nature 412, 198-202. Schwartz, H.T., and Horvitz, H.R. (2007). The C. elegans protein CEH-30 protects male-specific neurons from apoptosis independently of the Bcl-2 homolog CED-9. Genes Dev 21, 3181-3194. Shepelev, M.V., Chernoff, J., and Korobko, I.V. Rho family GTPase Chp/RhoV induces PC12 apoptotic cell death via JNK activation. Small Gtpases 2, 17-26. Sulston, J.E., and Horvitz, H.R. (1977). Post-embryonic cell lineages of the nematode, Caenorhabditis elegans. Dev Biol 56, 110-156. Sulston, J.E., Schierenberg, E., White, J.G., and Thomson, J.N. (1983). The Embryonic-Cell Lineage of the Nematode Caenorhabditis-Elegans. Dev Biol 100, 64-119. Walev, I., Reske, K., Palmer, M., Valeva, A., and Bhakdi, S. (1995). Potassium-inhibited processing of IL-1 beta in human monocytes. EMBO J 14, 1607-1614. Wang, X., Yang, C., Chai, J., Shi, Y., and Xue, D. (2002). Mechanisms of AIF-mediated apoptotic DNA degradation in Caenorhabditis elegans. Science 298, 1587-1592. Wiedau-Pazos, M., Goto, J.J., Rabizadeh, S., Gralla, E.B., Roe, J.A., Lee, M.K., Valentine, J.S., and Bredesen, D.E. (1996). Altered reactivity of superoxide dismutase in familial amyotrophic lateral sclerosis. Science 271, 515-518. Wu, Y.C., and Horvitz, H.R. (1998). The C. elegans cell corpse engulfment gene ced-7 encodes a protein similar to ABC transporters. Cell 93, 951-960. Yang, X., Chang, H.Y., and Baltimore, D. (1998). Essential role of CED-4 oligomerization in CED-3 activation and apoptosis. Science 281, 1355-1357. Yi, C.H., Sogah, D.K., Boyce, M., Degterev, A., Christofferson, D.E., and Yuan, J. (2007a). A genome-wide RNAi screen reveals multiple regulators of caspase activation. J Cell Biol 179, 619-626. Yi, C.H., Sogah, D.K., Boyce, M., Degterev, A., Christofferson, D.E., and Yuan, J. (2007b). A genome-wide RNAi screen reveals multiple regulators of caspase activation. J Cell Biol 179, 619-626. Yuan, J., and Horvitz, H.R. (1992). The Caenorhabditis elegans cell death gene ced-4 encodes a novel protein and is expressed during the period of extensive programmed cell death. Development 116, 309-320. Zhou, W., Simpson, P.J., McFadden, J.M., Townsend, C.A., Medghalchi, S.M., Vadlamudi, A., Pinn, M.L., Ronnett, G.V., and Kuhajda, F.P. (2003). Fatty acid synthase inhibition triggers apoptosis during S phase in human cancer cells. Cancer Res 63, 7330-7337. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45442 | - |
dc.description.abstract | 計畫性細胞凋亡是多細胞生物發育重要的過程, 而egl-1、ced-9、ced-4、ced-3這四個基因已經被發現是線蟲的計畫性細胞凋亡關鍵調節因子。當細胞決定死亡時,EGL-1會與CED-9結合,使得CED-4可以離開CED-9而活化CED-3,然而這些基因之調控的機制並不清楚。線蟲凋亡的細胞會產生不對稱分裂,所產生的兩個子細胞,較大的細胞會繼續分裂及分化,而較小的細胞則進行計畫性細胞凋亡。為了瞭解哪些基因影響計畫性細胞凋亡,所以我們實驗室設計了一組利用帶grp-1基因突變為背景之線蟲,篩選尾巴異常的蟲之方法。當grp-1或是與促進細胞凋亡的基因單獨突變時,線蟲只會有低比例的尾巴型態異常,然而當grp-1與促進細胞凋亡的基因同時突變時,則會使線蟲產生高比例的尾巴型態異常。藉由這樣的策略,已篩選出了一百多個線蟲突變株,其中三個為grp-1;tp27, grp-1;tp24, 以及 grp-1;tp172。這三個突變株有40~65%的尾巴型態異常,藉由DAPI染色,我發現這三個突變線蟲,在尾巴部分有多出的核,另外在胚胎1.5折時期,它們也有較少的細胞屍體。除此之外,grp-1;tp27 及grp-1;tp24有相當高的比例會有多的AVM或PVM細胞。除了藉由正向遺傳學的策略,我也利用了反向遺傳學的策略去尋找可能參與在計畫性細胞死亡的基因。之前兩篇文獻利用全基因組RNAi篩選的方式找出了在果蠅中可能參與於細胞凋亡的基因,參考兩篇文獻所指出的基因,我挑選了其中15個基因進行RNAi篩選實驗。藉由第一階段尾巴型態異常篩選,以及第二階段多出的Pn.aap細胞比例之篩選,我找出了四個候選基因--- K08F11.5, F10G8.9, C52D10.12, 及nkb-1。 | zh_TW |
dc.description.abstract | Programmed cell death is an important process during the development of multicellular organisms. In C. elegans, four important genes, egl-1, ced-9, ced-4, ced-3, have been shown involved in the core pathway to execute the programmed cell death. When the cell is decided to die, EGL-1 (BH3-contain protein) binds to CED-9/Bcl-2. Then, CED-4/Apaf-1 leaves from CED-9 and activate CED-3/caspase. In C. elegans, apoptotic cells are resulted from asymmetric cell division that give rise of two cells with different cell sizes and fates. The larger cell keeps dividing and differentiation, whereas the smaller one undergoes programmed cell death. However, how these genes are expressed and regulated remains largely unknown. To find new genes affecting the processes of programmed cell death, lab members performed a grp-1 enhancer screen. The screen was based on the observation that single mutation in grp-1 or any of the proapoptotic genes, egl-1, ced-4 and ced-3, resulted in a low penetrant of tail defect. But double mutations of grp-1 and any of the proapoptotic genes caused a high penetrance tail defect, which is likely attributed to an additional tail hypodermal cell. In this screen, three mutant strains grp-1;tp24, grp-1;tp27, and grp-1;tp172 were isolated. I found that these strains had moderate penetrance of tail defect. By staining with DAPI, I observed an extra nucleus in the tail region in these mutant worms. Three mutant strains are recessive and shows decreased number of cell corpses at the 1.5 fold embryo stage. In addition, grp-1;tp24 and grp-1;tp27 have higher percentage of the extra AVM and PVM phenotype than grp-1. These results show that these three mutants might involve in programmed cell death. In addition to forward genetic approaches, I also applied reverse genetic strategies to search for candidate genes that regulate programmed cell death. Recently two studies applied genome-wide RNA interference screen to identify regulators of apoptosis in Drosophila. According to these two studies 15 genes were chosen as my candidates. The grp-1 dependent tail defect analysis and subsequent analysis of extra surviving Pn.app cells, four genes, K08F11.5, F10G8.9, C52D10.12, and nkb-1, were found as candidates that might be involved in the programmed cell death. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T04:20:24Z (GMT). No. of bitstreams: 1 ntu-100-R98b43008-1.pdf: 1480828 bytes, checksum: 34c7869006460063688e21596b76fa1f (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | Table of contents
口試委員會審定書…………………………………………………………………. ii 誌謝………………………………………………………………………………. …iii 中文摘要………………………………………………………...…………… …. …iv Abstract…………………………………………………………...…………………. v Introduction……………………………………………….………...……………......1 Material and Methods……………..………………………………...…………….....5 Strains used……………………………………………………...………………...5 Isolation of new programmed cell death related genes………...…………………5 Microscopy………………………………………………...……………………..6 Cell corpse assay…………………………………………………..……………..6 Immunofluorescence staining and DAPI staining………………………………..7 Detection and analysis of touch neurons……………………..…………………..7 grp-1 dependency test……………………………………………..……………..8 Complementation test………………………………….…..………………….…8 Linkage group test………………………………………………………………..9 RNAi experiments……………………………………………………………..…9 Detection and analysis of Pn.aap cells……………………………….………….10 Results………………………………………………...…………………………....11 grp-1;tp24, grp-1;tp27, and grp-1;tp172 affect tail morphogenesis through a grp-1dependent manner………………………………………………….……..11 grp-1;tp24, grp-1;tp27, and grp-1;tp172 have different types of tail morphologic defect……………………………………………………………….…….……..13 grp-1;tp24, grp-1;tp27, and grp-1;tp172 have extra nuclei in the tail region…..15 grp-1;tp24, grp-1;tp27, and grp-1;tp172 have fewer corpses compared with wild type worms in the 1.5 fold stage………………………………...……….……...17 grp-1;tp24, and grp-1;tp27 have extra touch neurons…………...…………...…17 tp24 and tp27 complement each other and ced-8, while tp172 can complement ced-4………………………………………………………………...…………..18 tp27 maps to the chromosome X, while tp24 maps to the chromosome IV, and tp172 maps to the chromosome I……………………….…………..…………...19 Knockdown of 8 candidate genes cause significant tail defects………………...21 Knockdown of 4 candidate genes affect the numbers of extra Pn.aap cells…….22 Discussion……………………………………………………………..…………….25 Reference……………………………………………………………...…………….31 Tables and figures……………………………………………………..….…………37 Tabel 1. The corpse numbers and tail defect of mutant strains…..…………….37 Table 2. The grp-1 dependency test of grp-1;tp24……………………………..38 Table 3. The grp-1 dependency test of grp-1;tp27……………………………..39 Table 4. The grp-1 dependency test of grp-1;tp172…………………………....40 Table 5. The extra touch neurons of grp-1;tp24 and grp-1;tp27……………….41 Table 6. The complementation test results………………………………….….42 Table 7. X linkage group test results.. ……………………………….………...43 Table 8. tp24 linkage group test results.…………………………………..……44 Table 9. tp172 linkage group test results.………………………………………45 Table 10. Survival Pn.aap cells of candidate genes RNAi treated results……...46 Table 11. Survival Pn.aap cells of candidate genes RNAi treated results……47 Figure 1. The procedure of grp-1 dependency test…………………………….48 Figure 2. The tail defect morphology of grp-1; tp24. ………………………....49 Figure 3. The tail defect morphology of grp-1; tp27. ……………………...….50 Figure 4. The tail defect morphology of grp-1; tp172. …………………….….51 Figure 5. grp-1;tp24 and grp-1;tp27 have no extra cell border, while grp-1;tp172 has an extra cell border at the tail tip……………………….……….52 Figure 6. grp-1; tp24, grp-1; ;tp27, and grp-1; tp172 have an extra nucleus at tail region. ……………………………………………………….…...….53 | |
dc.language.iso | en | |
dc.title | 尋找在線蟲中參與計畫性細胞凋亡之新的調節子 | zh_TW |
dc.title | Identification of New Regulators Involved in
Programmed Cell Death | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李士傑,黃偉邦 | |
dc.subject.keyword | 線蟲,計畫性細胞凋亡,不對稱分裂,尾巴型態異常,細胞屍體, | zh_TW |
dc.subject.keyword | C. elegans,Programmed cell death,Asymmetric cell division,Tail defect morphology,cell corpses, | en |
dc.relation.page | 53 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-17 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-100-1.pdf 目前未授權公開取用 | 1.45 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。