Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes

作者：Daniel J. Klionsky, Hagai Abeliovich, Patrizia Agostinis, Devendra K. Agrawal, Gjumrakch Aliev, David S. Askew, Misuzu Baba, Eric H. Baehrecke, Ben A. Bahr, Andrea Ballabio, Bruce A. Bamber, Diane C. Bassham, Ettore Bergamini, Xiaoning Bi, Martine Biard‐Piechaczyk, Janice S. Blum, Dale E. Bredesen, Jeffrey L. Brodsky, John Hunter Brumell, Ulf T. Brunk, Wilfried Bursch, Nadine M. Camougrand, Eduardo Cebollero, Francesco Cecconi, Yingyu Chen, Lih‐Shen Chin, Augustine M.K. Choi, Charleen T. Chu, Jongkyeong Chung, Robert S. B. Clark, Peter G.H. Clarke, Steven Gerard Clarke, Corinne Clavé, John L. Cleveland, Patrice Codogno, María Isabel Colombo, Ana M. Coto‐Montes, James M. Cregg, Ana María Cuervo, Jayanta Debnath, Patrick B. Dennis, Phillip A. Dennis, Francesca Demarchi, Vojo P. Deretic, Rodney J. Devenish, Federica Di Sano, J. Fred Dice, Clark Distelhorst, Savithramma P. Dinesh‐Kumar, N. Tony Eissa, Marian DiFiglia, Mojgan Djavaheri‐Mergny, Frank C. Dorsey, Wulf Dröge, Michel Dron, William Augustine Dunn, Michael Duszenko, Zvulun Elazar, Audrey Esclatine, Eeva‐Liisa Eskelinen, László Fésüs, Kim D. Finley, José M. Fuentes, Juan Fueyo-Margareto, Kozo Fujisaki, Brigitte Galliot, Fen‐Biao Gao, David A. Gewirtz, Spencer Bruce Gibson, Antje Gohla, Alfred Lewis Goldberg, Ramon Gilberto Gonzalez, Cristina González‐Estévez, Sharon M. Gorski, Roberta A. Gottlieb, Dieter Häussinger, You-Wen He, Kim Heidenreich, Joseph A. Hill, Maria Høyer-Hansen, Xun Hu, Wei‐Pang Huang, Akiko Iwasaki, Marja Jäättelä, William T. Jackson, Xuejun Jiang, Shengkan V. Jin, Terje Johansen, Jae Ung Jung, Motoni Kadowaki, Chanhee Kang, Ameeta Kelekar, David H. Kessel, Jan A.K.W. Kiel, Hong Pyo Kim, Adi Kimchi, Timothy J. Kinsella, Kirill I. Kiselyov, Katsuhiko Kitamoto, Erwin Knecht, Masaaki Komatsu, Eiki Kominami, Seiji Kondo, Attila L. Kovács, Guido Kroemer, Chia-Yi Kuan, Rakesh Kumar, Mondira Kundu, Jacques Landry, Marianne Laporte, Weidong Le, Huan Lei, Beth Levine, Andrew P. Lieberman, Kah-Leong Lim, Fu-Cheng Lin, Willisa Liou, Leroy F. Liu, Gabriel Lopez‐Berestein, Carlos López-Otı́n, Bo Lü, Kay F. Macleod, Walter Malorni, Wim Martinet, Ken Matsuoka, Josef Mautner, Alfred J. Meijer, Alicia Meléndez, Paul A.M. Michels, Giovanni Miotto, Wilhelm Peter Mistiaen, Noboru Mizushima, Baharia Mograbi, Michael N. Moore, Paula Isabel Moreira, Yuji Moriyasu, T. Motyl, Christian Münz, Leon O. Murphy, Naweed I. Naqvi, Thomas P. Neufeld, Ichizo Nishino, Ralph A. Nixon, Takeshi Noda, Bernd Nürnberg, Michinaga Ogawa, Nancy L. Oleinick, Laura J. Olsen, Bülent Özpolat, Shoshana Paglin, Glen E. Palmer, Issidora S. Papassideri, Miles Parkes, David H. Perlmutter, George Perry, Mauro Piacentini, Ronit Pinkas‐Kramarski, Mark Prescott, Tassula Proikas-Cezanne, Nina Raben, Abdelhaq Rami, Fulvio Reggiori, Bärbel Rohrer, David Chaim Rubinsztein, Kevin M. Ryan, Junichi Sadoshima, Hiroshi Sakagami, Yasuyoshi Sakai, Marco Sandri, Chihiro Sasakawa, Miklós Sass, Claudio Schneider, Per O. Seglen, Oleksandr Seleverstov, Jeffrey E. Settleman, John J. Shacka, Irving M. Shapiro, Andrei A. Sibirny, Elaine C.M. Silva-Zacarín, Hans-Uwe Simon, Cristiano Simone, Anne Katrine Wrist Simonsen, Mark A. Smith, Katharina Spanel‐Borowski, Vickram Srinivas, Meredith A. Steeves, Harald Stenmark, Per Eivind STRØMHAUG, Carlos S. Subauste, Sugimoto Seiichiro, David L. Sulzer, Toshihiko Suzuki, Michele S. Swanson, Ira A. Tabas, Fumihiko Takeshita, Nicholas J. Talbot, Zsolt Tallóczy, Keiji Tanaka, Kozo Tanaka, Isei Tanida, Graham S. Taylor, J. Paul Taylor, Alexei Terman, Gianluca Tettamanti, Craig B. Thompson, Michael Thumm, Aviva M. Tolkovsky, Sharon A. Tooze, Ray Truant, Lesya V. Tumanovska, Yasuo Uchiyama, Takashi Ueno, Nestor Luis Uzcategui, Ida J. van der Klei, Eva Cristina Vaquero, Tibor Vellai, Michael W. Vogel, Hong-Gang Wang, Paul Christopher Webster, Zhijun Xi, Gutian Xiao, Joachim Yahalom, Jin-Ming Yang, George Yap, Xiao-Ming Yin, Tamotsu Yoshimori, Zhenyu Yue, Michisuke Yuzaki, Olga Zabirnyk, Xiaoxiang Zheng, Xiongwei Zhu, Russell L. Deter · 发表于：Autophagy · 年份：2008 · DOI：10.4161/auto.5338 · 被引用次数：2248 · 研究领域：Autophagy in Disease and Therapy、Microtubule and mitosis dynamics、Cellular transport and secretion

Research in autophagy continues to accelerate,(1) and 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.(2,3) There 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 ...