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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 楊維元(Wei Yuan Yang) | |
dc.contributor.author | Ting-Sung Hsieh | en |
dc.contributor.author | 謝廷松 | zh_TW |
dc.date.accessioned | 2021-06-13T16:42:50Z | - |
dc.date.available | 2016-07-26 | |
dc.date.copyright | 2011-07-26 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-07-17 | |
dc.identifier.citation | Andersson, S.B. (2008). Localization of a fluorescent source without numerical fitting. Opt Express 16, 18714-18724.
Annibale, P., Scarselli, M., Kodiyan, A., and Radenovic, A. (2010). Photoactivatable fluorescent protein mEos2 displays repeated photoactivation after a long-lived dark state in the red photoconverted form. J Phys Chem Lett 1, 1506-1510. Annibale, P., Vanni, S., Scarselli, M., Rothlisberger, U., and Radenovic, A. (2011). Identification of clustering artifacts in photoactivated localization microscopy. Nat Methods 8, 527-528. Axe, E.L., Walker, S.A., Manifava, M., Chandra, P., Roderick, H.L., Habermann, A., Griffiths, G., and Ktistakis, N.T. (2008). Autophagosome formation from membrane compartments enriched in phosphatidylinositol 3-phosphate and dynamically connected to the endoplasmic reticulum. J Cell Biol 182, 685-701. Axelrod, D. (1981). Cell-substrate contacts illuminated by total internal reflection fluorescence. J Cell Biol 89, 141-145. Axelrod, D., Burghardt, T.P., and Thompson, N.L. (1984). Total internal reflection fluorescence. Annu Rev Biophys Bioeng 13, 247-268. Behrends, C., Sowa, M.E., Gygi, S.P., and Harper, J.W. (2010). Network organization of the human autophagy system. Nature 466, 68-76. Betzig, E., Patterson, G.H., Sougrat, R., Lindwasser, O.W., Olenych, S., Bonifacino, J.S., Davidson, M.W., Lippincott-Schwartz, J., and Hess, H.F. (2006). Imaging intracellular fluorescent proteins at nanometer resolution. Science 313, 1642-1645. Betzig, E., and Trautman, J.K. (1992). Near-field optics: microscopy, spectroscopy, and surface modification beyond the diffraction limit. Science 257, 189-195. Bjorkoy, G., Lamark, T., Brech, A., Outzen, H., Perander, M., Overvatn, A., Stenmark, H., and Johansen, T. (2005). p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 171, 603-614. Chakrama, F.Z., Seguin-Py, S., Le Grand, J.N., Fraichard, A., Delage-Mourroux, R., Despouy, G., Perez, V., Jouvenot, M., and Boyer-Guittaut, M. (2010). GABARAPL1 (GEC1) associates with autophagic vesicles. Autophagy 6, 495-505. Chudakov, D.M., Lukyanov, S., and Lukyanov, K.A. (2007). Tracking intracellular protein movements using photoswitchable fluorescent proteins PS-CFP2 and Dendra2. Nat Protoc 2, 2024-2032. Clark, S.L., Jr. (1957). Cellular differentiation in the kidneys of newborn mice studies with the electron microscope. J Biophys Biochem Cytol 3, 349-362. Cuervo, A.M., and Dice, J.F. (1996). A receptor for the selective uptake and degradation of proteins by lysosomes. Science 273, 501-503. Cuervo, A.M., and Dice, J.F. (2000). Unique properties of lamp2a compared to other lamp2 isoforms. J Cell Sci 113, 4441-4450. Deter, R.L., and de Duve, C. (1967). Influence of glucagon, an inducer of cellular autophagy, on some physical properties of rat liver lysosomes. J Cell Biol 33, 437-449. Dice, J.F. (2007). Chaperone-mediated autophagy. Autophagy 3, 295-299. Drake, K.R., Kang, M., and Kenworthy, A.K. (2010). Nucleocytoplasmic distribution and dynamics of the autophagosome marker EGFP-LC3. PLoS ONE 5, e9806. English, L., Chemali, M., Duron, J., Rondeau, C., Laplante, A., Gingras, D., Alexander, D., Leib, D., Norbury, C., Lippe, R., et al. (2009). Autophagy enhances the presentation of endogenous viral antigens on MHC class I molecules during HSV-1 infection. Nat Immunol 10, 480-487. Eskelinen, E.L. (2008). To be or not to be? Examples of incorrect identification of autophagic compartments in conventional transmission electron microscopy of mammalian cells. Autophagy 4, 257-260. Filimonenko, M., Stuffers, S., Raiborg, C., Yamamoto, A., Malerod, L., Fisher, E.M., Isaacs, A., Brech, A., Stenmark, H., and Simonsen, A. (2007). Functional multivesicular bodies are required for autophagic clearance of protein aggregates associated with neurodegenerative disease. J Cell Biol 179, 485-500. Fujita, N., Hayashi-Nishino, M., Fukumoto, H., Omori, H., Yamamoto, A., Noda, T., and Yoshimori, T. (2008). An Atg4B mutant hampers the lipidation of LC3 paralogues and causes defects in autophagosome closure. Mol Biol Cell 19, 4651-4659. Geng, J., and Klionsky, D.J. (2008). The Atg8 and Atg12 ubiquitin-like conjugation systems in macroautophagy. EMBO Rep 9, 859-864. Glick, D., Barth, S., and Macleod, K.F. (2010). Autophagy: cellular and molecular mechanisms. J Pathol 221, 3-12. Gomes, L.C., Benedetto, G.D., and Scorrano, L. (2011). During autophagy mitochondria elongate, are spared from degradation and sustain cell viability. Nat Cell Biol 13, 589-598. Gordon, M.P., Ha, T., and Selvin, P.R. (2004). Single-molecule high-resolution imaging with photobleaching. Proc Natl Acad Sci USA 101, 6462-6465. Gould, T.J., Verkhusha, V.V., and Hess, S.T. (2009). Imaging biological structures with fluorescence photoactivation localization microscopy. Nat Protoc 4, 291-308. Gustafsson, M.G.L. (2000). Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J Microsc 198, 82-87. Hailey, D.W., Rambold, A.S., Satpute-Krishnan, P., Mitra, K., Sougrat, R., Kim, P.K., and Lippincott-Schwartz, J. (2010). Mitochondria supply membranes for autophagosome biogenesis during starvation. Cell 141, 656-667. Hayashi-Nishino, M., Fujita, N., Noda, T., Yamaguchi, A., Yoshimori, T., and Yamamoto, A. (2009). A subdomain of the endoplasmic reticulum forms a cradle for autophagosome formation. Nat Cell Biol 11, 1433-1437. Hedde, P.N., Fuchs, J., Oswald, F., Wiedenmann, J., and Nienhaus, G.U. (2009). Online image analysis software for photoactivation localization microscopy. Nat Methods 6, 689-690. Hell, S.W., and Wichmann, J. (1994). Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt Lett 19, 780-782. Henriques, R., Lelek, M., Fornasiero, E.F., Valtorta, F., Zimmer, C., and Mhlanga, M.M. (2010). QuickPALM: 3D real-time photoactivation nanoscopy image processing in ImageJ. Nat Methods 7, 339-340. Hess, S.T., Gould, T.J., Gudheti, M.V., Maas, S.A., Mills, K.D., and Zimmerberg, J. (2007). Dynamic clustered distribution of hemagglutinin resolved at 40 nm in living cell membranes discriminates between raft theories. Proc Natl Acad Sci USA 104, 17370-17375. Huang, B., Wang, W., Bates, M., and Zhuang, X. (2008). Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy. Science 319, 810-813. Ichimura, Y., Kirisako, T., Takao, T., Satomi, Y., Shimonishi, Y., Ishihara, N., Mizushima, N., Tanida, I., Kominami, E., Ohsumi, M., et al. (2000). A ubiquitin-like system mediates protein lipidation. Nature 408, 488-492. Itakura, E., and Mizushima, N. (2011). p62 targeting to the autophagosome formation site requires self-oligomerization but not LC3 binding. J Cell Biol 192, 17-27. Jahreiss, L., Menzies, F.M., and Rubinsztein, D.C. (2008). The itinerary of autophagosomes: from peripheral formation to kiss-and-run fusion with lysosomes. Traffic 9, 574-587. Juette, M.F., Gould, T.J., Lessard, M.D., Mlodzianoski, M.J., Nagpure, B.S., Bennett, B.T., Hess, S.T., and Bewersdorf, J. (2008). Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples. Nat Methods 5, 527-529. Kabeya, Y., Mizushima, N., Ueno, T., Yamamoto, A., Kirisako, T., Noda, T., Kominami, E., Ohsumi, Y., and Yoshimori, T. (2000). LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19, 5720-5728. Kabeya, Y., Mizushima, N., Yamamoto, A., Oshitani-Okamoto, S., Ohsumi, Y., and Yoshimori, T. (2004). LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci 117, 2805-2812. Kanki, T., Wang, K., Cao, Y., Baba, M., and Klionsky, D.J. (2009). Atg32 is a mitochondrial protein that confers selectivity during mitophagy. Dev Cell 17, 98-109. Kim, J., Huang, W.P., and Klionsky, D.J. (2001). Membrane recruitment of Aut7p in the autophagy and cytoplasm to vacuole targeting pathways requires Aut1p, Aut2p, and the autophagy conjugation complex. J Cell Biol 152, 51-64. Kirisako, T., Ichimura, Y., Okada, H., Kabeya, Y., Mizushima, N., Yoshimori, T., Ohsumi, M., Takao, T., Noda, T., and Ohsumi, Y. (2000). The reversible modification regulates the membrane-binding state of Apg8/Aut7 essential for autophagy and the cytoplasm to vacuole targeting pathway. J Cell Biol 151, 263-276. Klionsky, D.J. (2007). Autophagy: from phenomenology to molecular understanding in less than a decade. Nat Rev Mol Cell Biol 8, 931-937. Mann, S.S., and Hammarback, J.A. (1994). Molecular characterization of light chain 3. A microtubule binding subunit of MAP1A and MAP1B. J Biol Chem 269, 11492-11497. McKinney, S.A., Murphy, C.S., Hazelwood, K.L., Davidson, M.W., and Looger, L.L. (2009). A bright and photostable photoconvertible fluorescent protein. Nat Methods 6, 131-133. Mijaljica, D., Prescott, M., and Devenish, R.J. (2011). Microautophagy in mammalian cells: revisiting a 40-year-old conundrum. Autophagy 7, 1-10. Mizushima, N. (2007). Autophagy: process and function. Genes Dev 21, 2861-2873. Mizushima, N., Kuma, A., Kobayashi, Y., Yamamoto, A., Matsubae, M., Takao, T., Natsume, T., Ohsumi, Y., and Yoshimori, T. (2003). Mouse Apg16L, a novel WD-repeat protein, targets to the autophagic isolation membrane with the Apg12-Apg5 conjugate. J Cell Sci 116, 1679-1688. Mizushima, N., Noda, T., Yoshimori, T., Tanaka, Y., Ishii, T., George, M.D., Klionsky, D.J., Ohsumi, M., and Ohsumi, Y. (1998). A protein conjugation system essential for autophagy. Nature 395, 395-398. Mizushima, N., Yamamoto, A., Hatano, M., Kobayashi, Y., Kabeya, Y., Suzuki, K., Tokuhisa, T., Ohsumi, Y., and Yoshimori, T. (2001). Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J Cell Biol 152, 657-668. Monastyrska, I., Rieter, E., Klionsky, D.J., and Reggiori, F. (2009). Multiple roles of the cytoskeleton in autophagy. Biol Rev Camb Philos Soc 84, 431-448. Mortensen, K.I., Churchman, L.S., Spudich, J.A., and Flyvbjerg, H. (2010). Optimized localization analysis for single-molecule tracking and super-resolution microscopy. Nat Methods 7, 377-381. Morvan, J., Kochl, R., Watson, R., Collinson, L.M., Jefferies, H.B.J., and Tooze, S.A. (2009). In vitro reconstitution of fusion between immature autophagosomes and endosomes. Autophagy 5, 676-689. Nakatogawa, H., Ichimura, Y., and Ohsumi, Y. (2007). Atg8, a ubiquitin-like protein required for autophagosome formation, mediates membrane tethering and hemifusion. Cell 130, 165-178. Nakatogawa, H., Suzuki, K., Kamada, Y., and Ohsumi, Y. (2009). Dynamics and diversity in autophagy mechanisms: lessons from yeast. Nat Rev Mol Cell Biol 10, 458-467. Niu, L., and Yu, J. (2008). Investigating intracellular dynamics of FtsZ cytoskeleton with photoactivation single-molecule tracking. Biophys J 95, 2009-2016. Obara, K., Sekito, T., Niimi, K., and Ohsumi, Y. (2008). The Atg18-Atg2 complex is recruited to autophagic membranes via phosphatidylinositol 3-phosphate and exerts an essential function. J Biol Chem 283, 23972-23980. Okamoto, K., Kondo-Okamoto, N., and Ohsumi, Y. (2009). Mitochondria-anchored receptor Atg32 mediates degradation of mitochondria via selective autophagy. Dev Cell 17, 87-97. Orsi, A., Polson, H.E.J., and Tooze, S.A. (2010). Membrane trafficking events that partake in autophagy. Curr Opin Cell Biol 22, 150-156. Pankiv, S., Clausen, T.H., Lamark, T., Brech, A., Bruun, J.A., Outzen, H., Overvatn, A., Bjorkoy, G., and Johansen, T. (2007). p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 282, 24131-24145. Patterson, G.H., and Lippincott-Schwartz, J. (2002). A photoactivatable GFP for selective photolabeling of proteins and cells. Science 297, 1873-1877. Pavani, S.R., Thompson, M.A., Biteen, J.S., Lord, S.J., Liu, N., Twieg, R.J., Piestun, R., and Moerner, W.E. (2009). Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function. Proc Natl Acad Sci USA 106, 2995-2999. Pertsinidis, A., Zhang, Y., and Chu, S. (2010). Subnanometre single-molecule localization, registration and distance measurements. Nature 466, 647-651. Qu, X., Wu, D., Mets, L., and Scherer, N.F. (2004). Nanometer-localized multiple single-molecule fluorescence microscopy. Proc Natl Acad Sci USA 101, 11298-11303. Quan, T., Li, P., Long, F., Zeng, S., Luo, Q., Hedde, P.N., Nienhaus, G.U., and Huang, Z.L. (2010). Ultra-fast, high-precision image analysis for localization-based super resolution microscopy. Opt Express 18, 11867-11876. Ravikumar, B., Moreau, K., Jahreiss, L., Puri, C., and Rubinsztein, D.C. (2010). Plasma membrane contributes to the formation of pre-autophagosomal structures. Nat Cell Biol 12, 747-757. Razi, M., Chan, E.Y.W., and Tooze, S.A. (2009). Early endosomes and endosomal coatomer are required for autophagy. J Cell Biol 185, 305-321. Rust, M.J., Bates, M., and Zhuang, X. (2006). Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat Methods 3, 793-795. Sagiv, Y., Legesse-Miller, A., Porat, A., and Elazar, Z. (2000). GATE-16, a membrane transport modulator, interacts with NSF and the Golgi v-SNARE GOS-28. EMBO J 19, 1494-1504. Sahu, R., Kaushik, S., Clement, C.C., Cannizzo, E.S., Scharf, B., Follenzi, A., Potolicchio, I., Nieves, E., Cuervo, A.M., and Santambrogio, L. (2011). Microautophagy of cytosolic proteins by late endosomes. Dev Cell 20, 131-139. Sakai, Y., Koller, A., Rangell, L.K., Keller, G.A., and Subramani, S. (1998). Peroxisome degradation by microautophagy in Pichia pastoris: identification of specific steps and morphological intermediates. J Cell Biol 141, 625-636. Shintani, T., Suzuki, K., Kamada, Y., Noda, T., and Ohsumi, Y. (2001). Apg2p functions in autophagosome formation on the perivacuolar structure. J Biol Chem 276, 30452-30460. Shroff, H., Galbraith, C.G., Galbraith, J.A., White, H., Gillette, J., Olenych, S., Davidson, M.W., and Betzig, E. (2007). Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes. Proc Natl Acad Sci USA 104, 20308-20313. Shroff, H., White, H., and Betzig, E. (2008). Photoactivated localization microscopy (PALM) of adhesion complexes. Curr Protoc Cell Biol Chapter 4, Unit 4.21. Smith, C.S., Joseph, N., Rieger, B., and Lidke, K.A. (2010). Fast, single-molecule localization that achieves theoretically minimum uncertainty. Nat Methods 7, 373-375. Sternberg, S.R. (1983). Biomedical image processing. IEEE Comput 16, 22-34. Testa, I., Wurm, C.A., Medda, R., Rothermel, E., von Middendorf, C., Folling, J., Jakobs, S., Schonle, A., Hell, S.W., and Eggeling, C. (2010). Multicolor fluorescence nanoscopy in fixed and living cells by exciting conventional fluorophores with a single wavelength. Biophys J 99, 2686-2694. Thompson, R.E., Larson, D.R., and Webb, W.W. (2002). Precise nanometer localization analysis for individual fluorescent probes. Biophys J 82, 2775-2783. Tooze, S.A., and Yoshimori, T. (2010). The origin of the autophagosomal membrane. Nat Cell Biol 12, 831-835. Twig, G., Elorza, A., Molina, A.J., Mohamed, H., Wikstrom, J.D., Walzer, G., Stiles, L., Haigh, S.E., Katz, S., Las, G., et al. (2008). Fission and selective fusion govern mitochondrial segregation and elimination by autophagy. EMBO J 27, 433-446. Wang, C.W., Kim, J., Huang, W.P., Abeliovich, H., Stromhaug, P.E., Dunn, W.A., and Klionsky, D.J. (2001). Apg2 is a novel protein required for the cytoplasm to vacuole targeting, autophagy, and pexophagy pathways. J Biol Chem 276, 30442-30451. Wang, H., Bedford, F.K., Brandon, N.J., Moss, S.J., and Olsen, R.W. (1999). GABA(A)-receptor-associated protein links GABA(A) receptors and the cytoskeleton. Nature 397, 69-72. Weidberg, H., Shpilka, T., Shvets, E., Abada, A., Shimron, F., and Elazar, Z. (2011). LC3 and GATE-16 N termini mediate membrane fusion processes required for autophagosome biogenesis. Dev Cell 20, 444-454. Weidberg, H., Shvets, E., Shpilka, T., Shimron, F., Shinder, V., and Elazar, Z. (2010). LC3 and GATE-16/GABARAP subfamilies are both essential yet act differently in autophagosome biogenesis. EMBO J 29, 1792-1802. Yang, Z., and Klionsky, D.J. (2009). An overview of the molecular mechanism of autophagy. Curr Top Microbiol Immunol 335, 1-32. Yla-Anttila, P., Vihinen, H., Jokita, E., and Eskelinen, E.L. (2009). 3D tomography reveals connections between the phagophore and endoplasmic reticulum. Autophagy 5, 1180-1185. Youle, R.J., and Narendra, D.P. (2011). Mechanisms of mitophagy. Nat Rev Mol Cell Biol 12, 9-14. Zhang, B., Zerubia, J., and Olivo-Marin, J.C. (2007). Gaussian approximations of fluorescence microscope point-spread function models. Appl Opt 46, 1819-1829. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38706 | - |
dc.description.abstract | 在真核生物的演化歷程中,細胞自噬(巨型自噬)是一個高度保守的作用機制,它調控著經由溶小體所處理的巨分子或胞器分解。自噬作用在開始時,細胞質內會形成一個由脂質膜與多種蛋白構成的杯狀構造,叫做吞噬胞,它隨後會包圍細胞質中等待被分解的物質,最後封閉開口端產生一個由雙層脂質膜包覆的囊狀構造,稱為自噬體。在自噬體形成的過程中,把Atg8蛋白在哺乳動物細胞中的同源分子LC3和GABARAP接合到磷脂醯乙醇胺,以及隨後募集這些分子到自噬構造上是相當關鍵的步驟。目前在哺乳動物細胞中已經發現了四個LC3和四個GABARAP相關蛋白。近來的研究認為這些LC3和GABARAP蛋白可能分別在自噬體形成的不同階段裡作用,而且在吞噬胞的延拓或者吞噬目標物的選擇這兩個過程中具有獨特的功能。然而,迄今我們仍不清楚這些LC3/GABARAP蛋白究竟如何協同合作來促進自噬體形成,以及它們到底以怎樣的方式分布在延展中的吞噬胞上。在本篇研究中,我們利用光活化定位顯微術解析了細胞中的LC3/GABRAP蛋白分布至分子尺度。我們架構了光活化定位顯微術的儀器設備並開發了一套MATLAB程式組,Palmy PALM 803,來分析資料與繪製光活化定位顯微影像。到目前為止,透過單色光活化定位顯微術,我們已經揭露了細胞中mEos2-LC3B蛋白在可能為自噬構造上的分布狀態。進一步利用雙色或三色光活化定位顯微術,各種LC3/GABARAP分子間的相對位置也可以被決定。此外,細胞內的膜狀結構可以透過穿透式電子顯微技術來拍攝。結合穿透式電子顯微影像與透過光活化定位顯微術定出的LC3/GABARAP蛋白分布,我們將能對自噬體成熟進程中的膜狀結構動態以及各個LC3/GABARAP蛋白在自噬作用中的運作模式有更進一步的了解。 | zh_TW |
dc.description.abstract | Macroautophagy is a highly conserved intracellular process mediating lyso-some-dependent degradation of macromolecules or organelles. This process involves the formation of a cup-shaped membrane structure called phagophore, which subsequently engulfs selected cargos and matures into a double membrane-bound autophagosome. Conjugation of mammalian Atg8 homologues, LC3s and GABARAPs, to phosphatidylethanolamine (PE) and subsequent recruitment of LC3/GABARAP–PE to autophagic structures are crucial for autophagosome biogenesis. Currently in mammals there are four LC3 and four GABARAP variants identified. It has been suggested that LC3s and GABARAPs act at different stages in autophagosome formation and possibly play distinct roles in phagophore expansion or cargo selection control. Exactly how these LC3/GABARAP variants function cooperatively to achieve autophagosome formation and how they distribute on an expanding phagophore remain largely unknown. In this study, we employed photoactivated localization microscopy (PALM) to visualize LC3/GABARAP in vivo at molecular resolution. We set up PALM instrumentation and developed a suite of MATLAB scripts—Palmy PALM 803—for data analysis and image rendering. So far single-color PALM imaging has revealed mEos2-LC3B distributions on possible autophagic structures in the cell. Applying dual- or triple-color PALM, the respective localizations and the relative distributions of individual LC3/GABARAP can be determined. Intracellular membrane structures can also be imaged with transmission electron microscopy (TEM). Combining TEM images with identified LC3/GABARAP distributions by PALM, membrane dynamics through the autophagosome maturation and individual action modes of LC3/GABARAP variants in this autophagic process may be further clarified. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T16:42:50Z (GMT). No. of bitstreams: 1 ntu-100-R98B46001-1.pdf: 8471072 bytes, checksum: 77ee8ede1afe50d0eea789b118acb6c3 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | Acknowledgement .................................................................................... i
Abstract (in Chinese) .............................................................................. iii Abstract .................................................................................................... v I. Introduction .................................................................................... 1 I.1. An Overview of Autophagy ....................................................................... 1 I.2. The Process of Autophagy ....................................................................... 2 I.2.1. Phagophore Nucleation or Formation....................................................... 2 I.2.2. Atg5–Atg12 Conjugation ......................................................................... 3 I.2.3. Atg8 Processing and Its Membrane Integration ....................................... 4 I.2.4. Cargo Selection ......................................................................................... 5 I.2.5. Autophagosome Maturation and Its Fusion with Lysosome .................... 5 I.3. The Role of LC3/GABARAP in Phagophore Expansion and Cargo Selection............................................................................................................. 6 I.4. An Overview of Photoactivated Localization Microscopy ......................... 8 I.4.1. Biological Imaging Techniques and Super Resolution Microscopy......... 8 I.4.2. Photoactivated Localization Microscopy ............................................... 10 II. Materials and Procedures ........................................................... 12 II.1. Plasmid Construction ............................................................................. 12 II.2. Confocal Imaging ................................................................................... 13 II.3. PALM Instrumental Setup ...................................................................... 14 II.4. Coverslip Preparation ............................................................................ 14 II.5. PALM Imaging........................................................................................ 15 III. Software Development ................................................................ 18 III.1. Background Subtraction......................................................................... 19 III.2. Single Molecule Identification ................................................................ 20 III.3. Single Molecule Localization .................................................................. 24 III.4. Lateral Stage Drift Correction ................................................................ 26 III.5. Image Reconstruction ............................................................................ 28 IV. Results .......................................................................................... 30 IV.1. mEos2- and PS-CFP2-LC3/GABARAP Form Punctate Structures in the Cell…. .............................................................................................................. 30 IV.2. PALM Resolves Intriguing mEos2-LC3B Distributions and Possible Autophagic Structures ...................................................................................... 31 V. Discussions .................................................................................. 33 VI. Figures .......................................................................................... 36 VII. Tables ............................................................................................ 57 VIII. References .................................................................................... 60 IX. Appendices ................................................................................... 69 | |
dc.language.iso | en | |
dc.title | 解析細胞自噬相關構造上的 LC3/GABARAP 蛋白分布 | zh_TW |
dc.title | Mapping the LC3/GABARAP Distributions on Autophagic Structures with Photoactivated Localization Microscopy | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳光超(Guang-Chao Chen),陳培菱(Peilin Chen),王偉仲(Weichung Wang) | |
dc.subject.keyword | Atg8,LC3,GABARAP,細胞自噬,光活化定位顯微術, | zh_TW |
dc.subject.keyword | Atg8,LC3,GABARAP,autophagy,photoactivated localization microscopy, | en |
dc.relation.page | 69 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-07-17 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
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