المؤلفون: Nikolai Engedal, L. Gerner, Frank Sætre, Paula Szalai, Per Ottar Seglen, Morten Luhr

مصطلحات موضوعية: 0301 basic medicine, Autophagosome, Cell type, Endosome, Autophagy, Vacuole, Biology, Cell biology, Cell membrane, Cellular material, 03 medical and health sciences, Cytosol, 030104 developmental biology, medicine.anatomical_structure, medicine

الوصف: Autophagy (self-eating) is a common term for various processes by which cellular components are transferred to lysosomes for degradation. In macroautophagy, intracellular membrane structures termed "phagophores" expand to encapsulate autophagic cargo into sealed, double-membrane vacuoles termed "autophagosomes," which subsequently may fuse with endosomes to form intermediary vacuoles called "amphisomes," and finally with lysosomes to have their contents degraded and recycled. Autophagy is frequently analyzed by monitoring phagophore- and autophagosome-associated markers such as LC3. Although useful, it is becoming increasingly clear that very few, if any, of these marker proteins are entirely specific to the autophagic process. Moreover, phagophore/autophagosome markers cannot be used to measure autophagic activity since they are part of the autophagic machinery, or "cart," rather than autophagic cargo. Thus, there is a great need for functional assays in autophagy research. Here, we describe a method that quantitatively measures the nonselective autophagic sequestration of endogenous cytosolic cargo. The method is based on a crude separation of sedimentable cellular material from cytosol and a subsequent measurement of the fraction of a cytosolic enzyme activity transferred to the sedimentable fraction by autophagic sequestration. The original assay was first developed in 1990, but during the last few years we have systematically downscaled and simplified the method into the time- and cost-efficient procedure presented here, which can be performed with standard laboratory equipment and is suitable for any cell type.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::ff4d6a3344a7b5dbf96662a30d271461Test
https://doi.org/10.1016/bs.mie.2016.09.064Test

المؤلفون: Camilla Raiborg, Monica Fengsrud, Egil S. Erichsen, Trond Berg, Per Ottar Seglen

المصدر: European journal of cell biology. 79(12)

مصطلحات موضوعية: Autophagosome, Histology, Endosome, Biotin, Vacuole, Biology, Pathology and Forensic Medicine, Lysosome, Phagosomes, Organelle, medicine, Autophagy, Animals, Freeze Fracturing, Cryoelectron Microscopy, Membrane Proteins, Cell Biology, General Medicine, Intracellular Membranes, Transmembrane protein, Cell biology, Rats, Membrane, medicine.anatomical_structure, Membrane protein, Vacuoles

الوصف: The delimiting membranes of isolated autophagosomes from rat liver had extremely few transmembrane proteins, as indicated by the paucity of intramembrane particles in freeze-fracture images (about 20 particles/microm2, whereas isolated lysosomes had about 2000 particles/microm2). The autophagosomes also appeared to lack peripheral surface membrane proteins, since attempts to surface-biotinylate intact autophagosomes only yielded biotinylation of proteins from contaminating damaged mitochondria. All the membrane layers of multilamellar autophagosomes were equally particle-poor; the same was true of the autophagosome-forming, sequestering membrane complexes (phagophores). Isolated amphisomes (vacuoles formed by fusion between autophagosomes and endosomes) had more intramembrane particles than the autophagosomes (about 90 particles/microm2), and freeze-fracture images of these organelles frequently showed particle-rich endosomes fusing with particle-poor or particle-free autophagosomes. The appearence of multiple particle clusters suggested that a single autophagic vacuole could undergo multiple fusions with endosomes. Only the outermost membrane of bi- or multilamellar autophagic vacuoles appeared to engage in such fusions.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9ca1d291f557fe97bf580b3bb6fa8bfaTest
https://pubmed.ncbi.nlm.nih.gov/11152279Test

المؤلفون: Jan A.K.W. Kiel, Marianne M. Laporte, Yasuo Uchiyama, Lesya V. Tumanovska, Paula I. Moreira, Ray Truant, Russell L. Deter, Corinne Clavé, Xiongwei Zhu, Carlos López-Otín, Katharina Spanel-Borowski, Xuejun Jiang, David Kessel, William A. Dunn, Clark W. Distelhorst, Eeva-Liisa Eskelinen, Fu-Cheng Lin, Michele S. Swanson, Michael Thumm, You-Wen He, Alexei Terman, Leroy F. Liu, Hans-Uwe Simon, Marco Sandri, Thomas P. Neufeld, Per E. Stromhaug, Eva C. Vaquero, Janice S. Blum, Graham S. Taylor, David S. Askew, Giovanni Miotto, Michael Duszenko, Bruce A. Bamber, Marian DiFiglia, William T. Jackson, Katsuhiko Kitamoto, Bernd Nürnberg, J. Fred Dice, Chihiro Sasakawa, John J. Shacka, Attila L. Kovács, Kozo Fujisaki, Michael Moore, Savithramma P. Dinesh-Kumar, Ameeta Kelekar, David A. Gewirtz, Augustine M.K. Choi, Kah-Leong Lim, Bulent Ozpolat, Masaaki Komatsu, Kirill Kiselyov, Wilfried Bursch, Gianluca Tettamanti, Craig B. Thompson, Spencer B. Gibson, Takashi Ueno, Diane C. Bassham, Chanhee Kang, Mark A. Smith, Laura J. Olsen, Pyo Kim Hong, Michael J. Lenardo, Shengkan Jin, Patrick B. Dennis, Aviva M. Tolkovsky, Sharon M. Gorski, Gabriel Lopez-Berestein, Daniel J. Klionsky, Patrice Codogno, Antje Gohla, Hiroshi Sakagami, Terje Johansen, Yasuyoshi Sakai, Junichi Sadoshima, Takeshi Noda, José M. Fuentes, Michisuke Yuzaki, Mauro Piacentini, Michel Dron, Gutian Xiao, Frank C. Dorsey, Josef Mautner, Isei Tanida, Miklós Sass, John H. Brumell, Olga Zabirnyk, Francesco Cecconi, David C. Rubinsztein, Wim Martinet, Jeffrey Settleman, Sharon A. Tooze, Audrey Esclatine, John L. Cleveland, Kozo Tanaka, James M. Cregg, Jayanta Debnath, Leon Murphy, Oleksandr Seleverstov, Kay F. Macleod, Maria Høyer-Hansen, Naweed I. Naqvi, Ramon Gonzalez, Akiko Iwasaki, Ira Tabas, Patrizia Agostinis, Willisa Liou, Christian Münz, Michinaga Ogawa, Li Yu, Wulf Dröge, Huan Yao Lei, Baharia Mograbi, Meredith A. Steeves, Bo Lu, Marja Jäättelä, Ken Matsuoka, Ulf T. Brunk, Tassula Proikas-Cezanne, Robert S. B. Clark, Kevin M. Ryan, Harald Stenmark, Iryna Monastyrska, Jacques Landry, Ben A. Bahr, Mojgan Djavaheri-Mergny, Hong Gang Wang, Tomasz Motyl, Juan Fueyo, Vojo Deretic, Hagai Abeliovich, George S. Yap, László Fésüs, Paul A.M. Michels, John W. Wiley, Martine Biard-Piechaczyk, Xun Hu, Xiaoxiang Zheng, Wilhelm P. Mistiaen, Steven Clarke, Roberta A. Gottlieb, Kim A. Heidenreich, Seiji Kondo, Paul Webster, Fulvio Reggiori, Irving M. Shapiro, Carlos S. Subauste, Zsolt Tallóczy, Ichizo Nishino, Ettore Bergamini, Vickram Srinivas, Shoshana Paglin, Andrea Ballabio, Federica Di Sano, Nicholas J. Talbot, María Isabel Colombo, Zvulun Elazar, Dieter Häussinger, Chia Yi Kuan, Tibor Vellai, Anne Simonsen, Nancy L. Oleinick, Fen-Biao Gao, Jae U. Jung, Seiichiro Sugimoto, Gjumrakch Aliev, Abdelhaq Rami, Issidora S. Papassideri, Eduardo Cebollero, Beth Levine, N. Tony Eissa, David H. Perlmutter, Weidong Le, Wei-Pang Huang, Phillip A. Dennis, Joseph A. Hill, Michael W. Vogel, Charleen T. Chu, Noboru Mizushima, Walter Malorni, Brigitte Galliot, Andrew P. Lieberman, Erwin Knecht, Joachim Yahalom, Alfred L. Goldberg, Devendra K. Agrawal, Fumihiko Takeshita, Rodney J. Devenish, Keiji Tanaka, Lih-Shen Chin, Crisfiano Simone, Xiaoning Bi, David Sulzer, Zhijun Xi, Misuzu Baba, Andrei A. Sibirny, Elaine C.M. Silva-Zacarin, Guido Kroemer, Kim D. Finley, George Perry, Glen E. Palmer, Néstor L. Uzcátegui, Ana Coto-Montes, Ida J. van der Klei, Cristina González-Estévez, Xiao Ming Yin, Yingyu Chen, J. Paul Taylor, Alfred J. Meijer, Yuji Moriyasu, Jongkyeong Chung, Rakesh Kumar, Nadine Camougrand, Timothy J. Kinsella, Dale E. Bredesen, Bärbel Rohrer, Jeffrey L. Brodsky, Eiki Kominami, Ana Maria Cuervo, Adi Kimchi, Mondira Kundu, Peter G.H. Clarke, Zhenyu Yue, Ronit Pinkas-Kramarski, Per Ottar Seglen, Mark Prescott, Toshihiko Suzuki, Miles Parkes, Eric H. Baehrecke, Alicia Meléndez, Tamotsu Yoshimori, Claudio Schneider, Jin Ming Yang, Nina Raben, Francesca Demarchi, Ralph A. Nixon, Motoni Kadowaki

المصدر: Europe PubMed Central
Autophagy
Scopus-Elsevier
CIÊNCIAVITAE
ResearcherID
r-CIPF. Repositorio Institucional Producción Científica del Centro de Investigación Principe Felipe (CIPF)
instname
r-CIPF: Repositorio Institucional Producción Científica del Centro de Investigación Principe Felipe (CIPF)
Centro de Investigación Principe Felipe (CIPF)
Autophagy, vol. 4, no. 2, pp. 151-175
RIUR. Repositorio Institucional de la Universidad de La Rioja
Autophagy 4, 151-175 (2008)
Klionsky, D J, Abeliovich, H, Agostinis, P, Agrawal, D K, Aliev, G, Askew, D S, Baba, M, Baehrecke, E H, Bahr, B A, Ballabio, A, Bamber, B A, Bassham, D C, Bergamini, E, Bi, X, Biard-Piechaczyk, M, Blum, J S, Bredesen, D E, Brodsky, J L, Brumell, J H, Brunk, U T, Bursch, W, Camougrand, N, Cebollero, E, Cecconi, F, Chen, Y, Chin, L-S, Choi, A, Chu, C T, Chung, J, Clarke, P G H, Clark, R S B, Clarke, S G, Clavé, C, Cleveland, J L, Codogno, P, Colombo, M I, Coto-Montes, A, Cregg, J M, Cuervo, A M, Debnath, J, Demarchi, F, Dennis, P B, Dennis, P A, Deretic, V, Devenish, R J, Di Sano, F, Dice, J F, Difiglia, M, Dinesh-Kumar, S, Distelhorst, C W, Djavaheri-Mergny, M, Dorsey, F C, Dröge, W, Dron, M, Dunn, W A, Duszenko, M, Eissa, N T, Elazar, Z, Esclatine, A, Eskelinen, E-L, Fésüs, L, Finley, K D, Fuentes, J M, Fueyo, J, Fujisaki, K, Galliot, B, Gao, F-B, Gewirtz, D A, Gibson, S B, Gohla, A, Goldberg, A L, Gonzalez, R, González-Estévez, C, Gorski, S, Gottlieb, R A, Häussinger, D, He, Y-W, Heidenreich, K, Hill, J A, Høyer-Hansen, M, Hu, X, Huang, W-P, Iwasaki, A, Jäättelä, M, Jackson, W T, Jiang, X, Jin, S, Johansen, T, Jung, J U, Kadowaki, M, Kang, C, Kelekar, A, Kessel, D H, Kiel, J A K W, Kim, H P, Kimchi, A, Kinsella, T J, Kiselyov, K, Kitamoto, K, Knecht, E, Komatsu, M, Kominami, E, Kondo, S, Kovács, A L, Kroemer, G, Kuan, C-Y, Kumar, R, Kundu, M, Landry, J, Laporte, M, Le, W, Lei, H-Y, Lenardo, M J, Levine, B, Lieberman, A, Lim, K-L, Lin, F-C, Liou, W, Liu, L F, Lopez-Berestein, G, López-Otín, C, Lu, B, Macleod, K F, Malorni, W, Martinet, W, Matsuoka, K, Mautner, J, Meijer, A J, Meléndez, A, Michels, P, Miotto, G, Mistiaen, W P, Mizushima, N, Mograbi, B, Monastyrska, I, Moore, M N, Moreira, P I, Moriyasu, Y, Motyl, T, Münz, C, Murphy, L O, Naqvi, N I, Neufeld, T P, Nishino, I, Nixon, R A, Noda, T, Nürnberg, B, Ogawa, M, Oleinick, N L, Olsen, L J, Ozpolat, B, Paglin, S, Palmer, G E, Papassideri, I, Parkes, M, Perlmutter, D H, Perry, G, Piacentini, M, Pinkas-Kramarski, R, Prescott, M, Proikas-Cezanne, T, Raben, N, Rami, A, Reggiori, F, Rohrer, B, Rubinsztein, D C, Ryan, K M, Sadoshima, J, Sakagami, H, Sakai, Y, Sandri, M, Sasakawa, C, Sass, M, Schneider, C, Seglen, P O, Seleverstov, O, Settleman, J, Shacka, J J, Shapiro, I M, Sibirny, A, Silva-Zacarin, E C M, Simon, H-U, Simone, C, Simonsen, A, Smith, M A, Spanel-Borowski, K, Srinivas, V, Steeves, M, Stenmark, H, Stromhaug, P E, Subauste, C S, Sugimoto, S, Sulzer, D, Suzuki, T, Swanson, M S, Tabas, I, Takeshita, F, Talbot, N J, Tallóczy, Z, Tanaka, K, Tanaka, K, Tanida, I, Taylor, G S, Taylor, J P, Terman, A, Tettamanti, G, Thompson, C B, Thumm, M, Tolkovsky, A M, Tooze, S A, Truant, R, Tumanovska, L V, Uchiyama, Y, Ueno, T, Uzcátegui, N L, van der Klei, I, Vaquero, E C, Vellai, T, Vogel, M W, Wang, H-G, Webster, P, Wiley, J W, Xi, Z, Xiao, G, Yahalom, J, Yang, J-M, Yap, G, Yin, X-M, Yoshimori, T, Yu, L, Yue, Z, Yuzaki, M, Zabirnyk, O, Zheng, X, Zhu, X & Deter, R L 2008, ' Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes ', Autophagy, vol. 4, no. 2, pp. 151-75 . https://doi.org/10.4161/auto.5338Test

مصطلحات موضوعية: Autophagosome, GLUCAGON-INDUCED AUTOPHAGY, VINBLASTINE-INDUCED AUTOPHAGOCYTOSIS, STARVATION-INDUCED AUTOPHAGY, Autolysosome, autophagosome, Flux, Lysosome, Phagophore, Stress, Vacuole, MURINE PANCREATIC ACINAR, stress, ISOLATED RAT HEPATOCYTES, SEMINAL-VESICLE CELLS, DROSOPHILA FAT-BODY, BETAINE HOMOCYSTEINE METHYLTRANSFERASE, ELECTRON MICROSCOPIC EXAMINATION, CHAPERONE-MEDIATED AUTOPHAGY, 0302 clinical medicine, Chaperone-mediated autophagy, Phagosomes, Plants/metabolism, 0303 health sciences, Autophagy database, Plants, Saccharomyces cerevisiae Proteins/metabolism, autolysosome, Cell biology, Protein Transport, Eukaryotic Cells, Data Interpretation, Statistical, 030220 oncology & carcinogenesis, Microtubule-Associated Proteins/metabolism, lysosome, Autophagy/physiology, Microtubule-Associated Proteins, Phagosomes/metabolism, autophagy, Autophagy-Related Protein 8 Family, Saccharomyces cerevisiae Proteins, Settore BIO/06, Guidelines as Topic, Computational biology, Biology, Models, Biological, Article, 03 medical and health sciences, Animals, Humans, Set (psychology), Molecular Biology, Microscopy, Fluorescence/methods, phagophore, 030304 developmental biology, Eukaryotic Cells/physiology, flux, vacuole, Interpretation (logic), Clinical Laboratory Techniques, Laboratory Techniques and Procedures, Phagosomes/physiology, Protein Processing, Post-Translational, Autophagy, Cell Biology, Microscopy, Fluorescence, Human medicine

الوصف: Research in autophagy continues to accelerate,1and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.2,3There are many useful and convenient methods that can be used to monitor macroautophagy in yeast, but relatively few in other model systems, and there is much confusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from fully functional autophagy that includes delivery to, and degradation within, lysosomes (in most higher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for the selection and interpretation of the methods that can be used by investigators who are attempting to examine macroautophagy and related processes, as well as by reviewers who need to provide realistic and reasonable critiques of papers that investigate these processes. This set of guidelines is not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to verify an autophagic response. ©2008 Landes Bioscience.

وصف الملف: pdf; application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7c3f52fead4a8312d44d281412603b27Test
http://europepmc.org/abstract/med/18188003Test