請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69088
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 吳瑞菁 | |
dc.contributor.author | Shang-Yang Chen | en |
dc.contributor.author | 陳尚暘 | zh_TW |
dc.date.accessioned | 2021-06-17T02:51:18Z | - |
dc.date.available | 2027-08-15 | |
dc.date.copyright | 2017-09-13 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-08-15 | |
dc.identifier.citation | 1. Jallepalli, P.V. and C. Lengauer, Chromosome segregation and cancer: cutting through the mystery. Nat Rev Cancer, 2001. 1(2): p. 109-17.
2. Millband, D.N., L. Campbell, and K.G. Hardwick, The awesome power of multiple model systems: interpreting the complex nature of spindle checkpoint signaling. Trends in Cell Biology, 2002. 12(5): p. 205-209. 3. Sudakin, V., G.K.T. Chan, and T.J. Yen, Checkpoint inhibition of the APC/C in HeLa cells is mediated by a complex of BUBR1, BUB3, CDC20, and MAD2. The Journal of Cell Biology, 2001. 154(5): p. 925-936. 4. Musacchio, A. and E.D. Salmon, The spindle-assembly checkpoint in space and time. Nat Rev Mol Cell Biol, 2007. 8(5): p. 379-393. 5. Manchado, E., M. Eguren, and M. Malumbres, The anaphase-promoting complex/cyclosome (APC/C): cell-cycle-dependent and -independent functions. Biochemical Society Transactions, 2010. 38(1): p. 65. 6. Haering, C.H., et al., The cohesin ring concatenates sister DNA molecules. Nature, 2008. 454(7202): p. 297-301. 7. Hornig, N.C.D., et al., The Dual Mechanism of Separase Regulation by Securin. Current Biology, 2002. 12(12): p. 973-982. 8. Uhlmann, F., F. Lottspeich, and K. Nasmyth, Sister-chromatid separation at anaphase onset is promoted by cleavage of the cohesin subunit Scc1. Nature, 1999. 400(6739): p. 37-42. 9. Musacchio, A. and K.G. Hardwick, The spindle checkpoint: structural insights into dynamic signalling. Nat Rev Mol Cell Biol, 2002. 3(10): p. 731-741. 10. Dumont, J. and A. Desai, Acentrosomal spindle assembly and chromosome segregation during oocyte meiosis. Trends in Cell Biology, 2012. 22(5): p. 241-249. 11. Sharpe, R.M., et al., Proliferation and functional maturation of Sertoli cells, and their relevance to disorders of testis function in adulthood. Reproduction, 2003. 125(6): p. 769-784. 12. Shaha, C., R. Tripathi, and D.P. Mishra, Male germ cell apoptosis: regulation and biology. Philosophical Transactions of the Royal Society B: Biological Sciences, 2010. 365(1546): p. 1501-1515. 13. Shakes, D.C., et al., Spermatogenesis-Specific Features of the Meiotic Program in Caenorhabditis elegans. PLOS Genetics, 2009. 5(8): p. e1000611. 14. Gartner, A., P.R. Boag, and T.K. Blackwell, Germline survival and apoptosis. WormBook, 2008: p. 1-20. 15. Stiernagle, T., Maintenance of C. elegans. WormBook, 2006: p. 1-11. 16. Sun, S.-C. and N.-H. Kim, Spindle assembly checkpoint and its regulators in meiosis. Human Reproduction Update, 2012. 18(1): p. 60-72. 17. Wassmann, K., T. Niault, and B. Maro, Metaphase I Arrest upon Activation of the Mad2-Dependent Spindle Checkpoint in Mouse Oocytes. Current Biology, 2003. 13(18): p. 1596-1608. 18. McNally, K.L. and F.J. McNally, Fertilization initiates the transition from anaphase I to metaphase II during female meiosis in C. elegans. Developmental Biology, 2005. 282(1): p. 218-230. 19. Thornton, B.R. and D.P. Toczyski, Securin and B-cyclin/CDK are the only essential targets of the APC. Nat Cell Biol, 2003. 5(12): p. 1090-1094. 20. Nasmyth, K., Disseminating the Genome: Joining, Resolving, and Separating Sister Chromatids During Mitosis and Meiosis. Annual Review of Genetics, 2001. 35(1): p. 673-745. 21. Davis, E.S., et al., Multiple subunits of the Caenorhabditis elegans anaphase-promoting complex are required for chromosome segregation during meiosis I. Genetics, 2002. 160(2): p. 805-813. 22. Stein, K.K., et al., Components of the Spindle Assembly Checkpoint Regulate the Anaphase-Promoting Complex During Meiosis in Caenorhabditis elegans. Genetics, 2007. 175(1): p. 107-123. 23. Shah, J.V., et al., Dynamics of Centromere and Kinetochore Proteins: Implications for Checkpoint Signaling and Silencing. Current Biology, 2004. 14(11): p. 942-952. 24. Collin, P., et al., The Spindle Assembly Checkpoint works like a rheostat not a toggle-switch. Nature cell biology, 2013. 15(11): p. 10.1038/ncb2855. 25. Golden, A., et al., Metaphase to Anaphase (mat) Transition–Defective Mutants inCaenorhabditis elegans. The Journal of Cell Biology, 2000. 151(7): p. 1469-1482. 26. Nystul, T.G., et al., Suspended Animation in C. elegans Requires the Spindle Checkpoint. Science, 2003. 302(5647): p. 1038. 27. Kitajima, T.S., et al., Shugoshin collaborates with protein phosphatase 2A to protect cohesin. Nature, 2006. 441(7089): p. 46-52. 28. Salah, S.-M. and K. Nasmyth, Destruction of the securin Pds1p occurs at the onset of anaphase during both meiotic divisions in yeast. Chromosoma, 2000. 109(1): p. 27-34. 29. Hached, K., et al., Mps1 at kinetochores is essential for female mouse meiosis I. Development, 2011. 138(11): p. 2261. 30. Homer, H.A., et al., Mad2 is required for inhibiting securin and cyclin B degradation following spindle depolymerisation in meiosis I mouse oocytes. Reproduction, 2005. 130(6): p. 829-843. 31. Touati, S.A., et al., Mouse oocytes depend on BubR1 for proper chromosome segregation but not for prophase I arrest. Nature communications, 2015. 6: p. 6946-6946. 32. Liu, D., et al., Meiosis I in Xenopus oocytes is not error-prone despite lacking spindle assembly checkpoint. Cell Cycle, 2014. 13(10): p. 1602-1606. 33. Batiha, O. and A. Swan, Evidence that the spindle assembly checkpoint does not regulate APCFzy activity in Drosophila female meiosis. Genome, 2011. 55(1): p. 63-67. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69088 | - |
dc.description.abstract | 細胞分裂對於多細胞生物的成長、再生及生育都極為重要。在有絲分裂的系統中,從細胞分裂中期過渡到分裂後期的過程是由紡錘體檢查點SAC(spoindle assembly checkpoint)所嚴格的控管。SAC經由抑制一種E3泛素連接酶APC/C,來防止姊妹染色體之間的黏連蛋白被降解以及染色體分離。但是在雄性減數分裂系統中,細胞只進行一次染色體複製接著連續兩次染色體分離,而目前對於這兩次染色體分離的調控機制是否由SAC所調控尚不清楚。
為了研究雄性減數分裂的過程是否有SAC的參與,我們利用一個演算法來自動且快速分析我們所拍攝的縮時攝影影像中,位於染色體周圍的蛋白質表現量。我們發現位於著絲點外層,同時也是SAC訊號的蛋白BUB-1在第一次染色體分離時會脫離染色體,接著在第二次分裂中期開始之前會回到染色體上,表示著絲點的結構,同時也是SAC訊號的模板會在兩次減數分裂之間有重新再構築的動作。相反的,APC/C的作用目標securin雖然會在第一次染色體分離時被分解,但在第二次染色體分裂的過程中並不會重新聚集回染色體附近,推論第二次染色體分離的調控不需要APC/C的參與。為了更做進一步驗證,我們對精母細胞施予蛋白酶體抑制劑,發現初級精母細胞會被抑制在第一次分裂中期,然而二級精母細胞能夠完成第二次的染色體及細胞分離,推論第二次染色體分離的調控和第一次分裂不同,不需要蛋白酶體的參與。 作為總結,我們認為對有絲分裂的系統而言相當重要的APC/C以及蛋白酶體系統,不存在於第二次減數分裂的調控機制中。 | zh_TW |
dc.description.abstract | Cell division is important in all multicellular organisms for vital processes including growth, regeneration and reproduction. During cell cycle, the transition from metaphase to anaphase strictly controlled by Spindle Assembly Checkpoint (SAC). Until all chromosomes are properly attached by spindle microtubules, SAC prevents chromosome separation by inhibiting APC/C, an E3 ubiquitin ligase essential for releasing sister chromatid cohesion. In male meiosis, however, two consecutive chromosome separation events occur after one round of chromosome duplication. Thus far, it is unclear if both male meiotic divisions subjected to the control of canonical SAC signaling. To investigate if SAC signals function during male meiotic divisions, we developed an algorithm that allows automatic quantification of the levels of chromosome-associated proteins through the divisions in time-lapse recordings. We found that outer kinetochore SAC signaling protein BUB-1 released from chromosomes during first chromosome segregation event and recruited back to chromosome before second chromosome segregation takes place, indicating proper kinetochore structure and platform for SAC disassembled and re-assembled between two divisions. Contrarily, securin, the direct target of APC/C activity, though is degraded when first chromosome separation is initiated, fails to be recruited to chromosome during second division. These results indicate that the second male meiotic division does not required APC/C and the proteasome-dependent protein degradation. To test this, we examined the progression of division in primary and secondary spermatocytes treated with proteasome inhibitor MG132. As expected, primary spermatocytes treated with MG132 were stalled at metaphase I. Interestingly, secondary spermatocytes treated with MG132 were able to complete chromosome segregation and division. These results suggest the second male meiotic division might regulated differently compared to the first meiosis. Taken together, we hypothesize that the APC/C-proteasome system, which is crucial for canonical SAC signaling, does not participate in second chromosome segregation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T02:51:18Z (GMT). No. of bitstreams: 1 ntu-106-R04424021-1.pdf: 1844953 bytes, checksum: 0d4ab6349e535939b40e140f0826edd8 (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 謝詞 i
摘要 ii Abstract iii Table of content v Background 1 Material and Method 5 Strains 5 Temperature-shift assay 5 Time-laps recording 5 Image processing and statistical analysis 6 Results 7 Hypothetical models for SAC activities during male meiotic divisions 7 Automatic quantification of the levels of SAC-related proteins 8 Inner kinetochore protein KNL-3 localizes around chromosomes through entire meiotic division event 8 Outer kinetochore protein BUB-1 is re-established after first meiotic division 9 IFY-1 quickly degraded during first meiosis without recovery in second meiosis 10 APC/C activity involves in first male meiotic division 11 Proteasome activity is crucial for first but not second meiotic division 13 There is an APC/C-proteasome independent system for securin degradation after NEBD 14 Discussion 17 Inner kinetochore structure preserves between two chromosome segregation events 17 Besides canonical APC/C-proteasome pathway, there are other mechanisms that cause securin degradation 18 Canonical SAC system control first meiotic division 19 Second meiotic division is not regulated by canonical SAC model 19 Do SAC exist in male and female meiosis and is it indispensable? 21 Reference 22 Figures 27 Figure 1. SAC system in the regulation of mitotic progression 27 Figure 2. Hypothetical models for SAC activities during male meiotic divisions. 28 Figure 3. Inner kinetochore protein KNL3 localizes around chromosomes during entire meiotic division event 30 Figure 4. Kinetics of the outer kinetochore protein BUB-1 during meiosis I and meiosis II 32 Figure 5. IFY-1 quickly degraded during first meiosis without recovery in second meiosis 34 Figure 6. APC/C activity involves in first male meiotic division 36 Figure 7. Proteasome activity is crucial for first meiotic division but not in second meiotic division 38 Figure 8. Securin degradation event is not regulated only by APC/C-proteasome system 39 Tables 40 Table 1. Strains used in this study 40 | |
dc.language.iso | en | |
dc.title | 利用線蟲為模型探討紡錘體檢查點在調控精子生成時的角色 | zh_TW |
dc.title | Investigating the Spindle Assembly Checkpoint Regulation in C. elegans Spermatogenesis | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 吳益群,潘俊良,蘇剛毅 | |
dc.subject.keyword | 紡錘體檢查點,雄性減數分裂,縮時攝影,泛素連接?,蛋白?體, | zh_TW |
dc.subject.keyword | SAC,male meiosis,time-lapse recording,APC/C,proteasome, | en |
dc.relation.page | 40 | |
dc.identifier.doi | 10.6342/NTU201701440 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-08-15 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
顯示於系所單位: | 醫學檢驗暨生物技術學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-106-1.pdf 目前未授權公開取用 | 1.8 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。