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Guidelines for the use and interpretation of assays for monitoring autophagy

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Klionsky, Daniel J.;Abdalla, Fabio C.;Abeliovich, Hagai;Abraham, Robert T.;Acevedo-Arozena, Abraham;Adeli, Khosrow;Agholme, Lotta;Agnello, Maria;Agostinis, Patrizia;Aguirre-Ghiso, Julio A.;Ahn, Hyung Jun;Ait-Mohamed, Ouardia;Ait-Si-Ali, Slimane;Akematsu, Takahiko;Akira, Shizuo;Al-Younes, Hesham M.;Al-Zeer, Munir A.;Albert, Matthew L.;Albin, Roger L.;Alegre-Abarrategui, Javier;Aleo, Maria Francesca;Alirezaei, Mehrdad;Almasan, Alexandru;Almonte-Becerril, Maylin;Amano, Atsuo;Amaravadi, Ravi;Amarnath, Shoba;Amer, Amal O.;Andrieu-Abadie, Nathalie;Anantharam, Vellareddy;Ann, David K.;Anoopkumar-Dukie, Shailendra;Aoki, Hiroshi;Apostolova, Nadezda;Arancia, Giuseppe;Aris, John P.;Asanuma, Katsuhiko;Asare, Nana Y. O.;Ashida, Hisashi;Askanas, Valerie;Askew, David S.;Auberger, Patrick;Baba, Misuzu;Backues, Steven K.;Baehrecke, Eric H.;Bahr, Ben A.;Bai, Xue-Yuan;Bailly, Yannick;Baiocchi, Robert;Baldini, Giulia;Balduini, Walter;Ballabio, Andrea;Bamber, Bruce A.;Bampton, Edward T. W.;Banhegyi, Gabor;Bartholomew, Clinton R.;Bassham, Diane C.;Bast, Robert C., Jr.;Batoko, Henri;Bay, Boon-Huat;Beau, Isabelle;Bechet, Daniel M.;Begley, Thomas J.;Behl, Christian;Behrends, Christian;Bekri, Soumeya;Bellaire, Bryan;Bendall, Linda J.;Benetti, Luca;Berliocchi, Laura;Bernardi, Henri;Bernassola, Francesca;Besteiro, Sebastien;Bhatia-Kissova, Ingrid;Bi, Xiaoning;Biard-Piechaczyk, Martine;Blum, Janice S.;Boise, Lawrence H.;Bonaldo, Paolo;Boone, David L.;Bornhauser, Beat C.;Bortoluci, Karina R.;Bossis, Ioannis;Bost, Frederic;Bourquin, Jean-Pierre;Boya, Patricia;Boyer-Guittaut, Michael;Bozhkov, Peter V.;Brady, Nathan R.;Brancolini, Claudio;Brech, Andreas;Brenman, Jay E.;Brennand, Ana;Bresnick, Emery H.;Brest, Patrick;Bridges, Dave;Bristol, Molly L.;Brookes, Paul S.;Brown, Eric J.;Brumell, John H.;Brunetti-Pierri, Nicola;Brunk, Ulf T.;Bulman, Dennis E.;Bultman, Scott J.;Bultynck, Greet;Burbulla, Lena F.;Bursch, Wilfried;Butchar, Jonathan P.;Buzgariu, Wanda;Bydlowski, Sergio P.;Cadwell, Ken;Cahova, Monika;Cai, Dongsheng;Cai, Jiyang;Cai, Qian;Calabretta, Bruno;Calvo-Garrido, Javier;Camougrand, Nadine;Campanella, Michelangelo;Campos-Salinas, Jenny;Candi, Eleonora;Cao, Lizhi;Caplan, Allan B.;Carding, Simon R.;Cardoso, Sandra M.;Carew, Jennifer S.;Carlin, Cathleen R.;Carmignac, Virgine;Carneiro, Leticia A. M.;Carra, Serena;Caruso, Rosario A.;Casari, Giorgio;Casas, Caty;Castino, Roberta;Cebollero, Eduardo;Cecconi, Francesco;Celli, Jean;Chaachouay, Hassan;Chae, Han-Jung;Chai, Chee-Yin;Chan, David C.;Chan, Edmond Y.;Chang, Raymond Chuen-Chung;Che, Chi-Ming;Chen, Ching-Chow;Chen, Guang-Chao;Chen, Guo-Qiang;Chen, Min;Chen, Quan;Chen, Steve S. -L.;Chen, WenLi;Chen, Xi;Chen, Xiangmei;Chen, Xiequn;Chen, Ye-Guang;Chen, Yingyu;Chen, Yongqiang;Chen, Yu-Jen;Chen, Zhixiang;Cheng, Alan;Cheng, Christopher H. K.;Cheng, Yan;Cheong, Heesun;Cheong, Jae-Ho;Cherry, Sara;Chess-Williams, Russ;Cheung, Zelda H.;Chevet, Eric;Chiang, Hui-Ling;Chiarelli, Roberto;Chiba, Tomoki;Chin, Lih-Shen;Chiou, Shih-Hwa;Chisari, Francis V.;Cho, Chi Hin;Cho, Dong-Hyung;Choi, Augustine M. K.;Choi, DooSeok;Choi, Kyeong Sook;Choi, Mary E.;Chouaib, Salem;Choubey, Divaker;Choubey, Vinay;Chu, Charleen T.;Chuang, Tsung-Hsien;Chueh, Sheau-Huei;Chun, Taehoon;Chwae, Yong-Joon;Chye, Mee-Len;Ciarcia, Roberto;Ciriolo, Maria R.;Clague, Michael J.;Clark, Robert S. B.;Clarke, Peter G. H.;Clarke, Robert;Codogno, Patrice;Coller, Hilary A.;Colombo, Maria I.;Comincini, Sergio;Condello, Maria;Condorelli, Fabrizio;Cookson, Mark R.;Coppens, Graham H. Coombs Isabelle;Corbalan, Ramon;Cossart, Pascale;Costelli, Paola;Costes, Safia;Coto-Montes, Ana;Couve, Eduardo;Coxon, Fraser P.;Cregg, James M.;Crespo, Jose L.;Cronje, Marianne J.;Cuervo, Ana Maria;Cullen, Joseph J.;Czaja, Mark J.;D'Amelio, Marcello;Darfeuille-Michaud, Arlette;Davids, Lester M.;Davies, Faith E.;De Felici, Massimo;de Groot, John F.;de Haan, Cornelis A. M.;De Martino, Luisa;De Milito, Angelo;De Tata, Vincenzo;Debnath, Jayanta;Degterev, Alexei;Dehay, Benjamin;Delbridge, Lea M. 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Wade;Harris, Adrian L.;Harris, James;Harris, Steven D.;Hashimoto, Makoto;Haspel, Jeffrey A.;Hayashi, Shin-ichiro;Hazelhurst, Lori A.;He, Congcong;He, You-Wen;Hebert, Marie-Josee;Heidenreich, Kim A.;Helfrich, Miep H.;Helgason, Gudmundur V.;Henske, Elizabeth P.;Herman, Brian;Herman, Paul K.;Hetz, Claudio;Hilfiker, Sabine;Hill, Joseph A.;Hocking, Lynne J.;Hofman, Paul;Hofmann, Thomas G.;Hoehfeld, Joerg;Holyoake, Tessa L.;Hong, Ming-Huang;Hood, David A.;Hotamisligil, Goekhan S.;Houwerzijl, Ewout J.;Hoyer-Hansen, Maria;Hu, Bingren;Hu, Chien-An A.;Hu, Hong-Ming;Hua, Ya;Huang, Canhua;Huang, Ju;Huang, Shengbing;Huang, Wei-Pang;Huber, Tobias B.;Huh, Won-Ki;Hung, Tai-Ho;Hupp, Ted R.;Hur, Gang Min;Hurley, James B.;Hussain, Sabah N. 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M.;Michiels, Carine;Mijaljica, Dalibor;Milojkovic, Ana;Minucci, Saverio;Miracco, Clelia;Miranti, Cindy K.;Mitroulis, Ioannis;Miyazawa, Keisuke;Mizushima, Noboru;Mograbi, Baharia;Mohseni, Simin;Molero, Xavier;Mollereau, Bertrand;Mollinedo, Faustino;Momoi, Takashi;Monastyrska, Iryna;Monick, Martha M.;Monteiro, Mervyn J.;Moore, Michael N.;Mora, Rodrigo;Moreau, Kevin;Moreira, Paula I.;Moriyasu, Yuji;Moscat, Jorge;Mostowy, Serge;Mottram, Jeremy C.;Motyl, Tomasz;Moussa, Charbel E. -H.;Mueller, Sylke;Muenger, Karl;Muenz, Christian;Murphy, Leon O.;Murphy, Maureen E.;Musaro, Antonio;Mysorekar, Indira;Nagata, Eiichiro;Nagata, Kazuhiro;Nahimana, Aimable;Nair, Usha;Nakagawa, Toshiyuki;Nakahira, Kiichi;Nakano, Hiroyasu;Nakataogawa, Hitoshi;Nanjundan, Meera;Naqvi, Naweed I.;Narendra, Derek P.;Narita, Masashi;Navarro, Miguel;Nawrocki, Steffan T.;Nazarko, Taras Y.;Nemchenko, Andriy;Netea, Mihai G.;Neufeld, Thomas P.;Ney, Paul A.;Nezis, Ioannis P.;Huu Phuc Nguyen;Nie, Daotai;Nishino, Ichizo;Nislow, Corey;Nixon, Ralph A.;Noda, Takeshi;Noegel, Angelika A.;Nogalska, Anna;Noguchi, Satoru;Notterpek, Lucia;Novak, Ivana;Nozaki, Tomoyoshi;Nukina, Nobuyuki;Nuernberger, Thorsten;Nyfeler, Beat;Obara, Keisuke;Oberley, Terry D.;Oddo, Salvatore;Ogawa, Michinaga;Ohashi, Toya;Okamoto, Koji;Oleinick, Nancy L.;Oliver, F. 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Ashutosh;Rathmell, Jeffrey C.;Ravikumar, Brinda;Ray, Swapan K.;Reed, Bruce H.;Reed, John C.;Reggiori, Fulvio;Regnier-Vigouroux, Anne;Reichert, Andreas S.;Reiners, John J., Jr.;Reiter, Russel J.;Ren, Jun;Revuelta, Jose L.;Rhodes, Christopher J.;Ritis, Konstantinos;Rizzo, Elizete;Robbins, Jeffrey;Roberge, Michel;Roca, Hernan;Roccheri, Maria C.;Rocchi, Stephane;Rodemann, H. Peter;de Cordoba, Santiago Rodriguez;Rohrer, Baerbel;Roninson, Igor B.;Rosen, Kirill;Rost-Roszkowska, Magdalena M.;Rouis, Mustapha;Rouschop, Kasper M. A.;Rovetta, Francesca;Rubin, Brian P.;Rubinsztein, David C.;Ruckdeschel, Klaus;Rucker, Edmund B., III;Rudich, Assaf;Rudolf, Emil;Ruiz-Opazo, Nelson;Russo, Rossella;Rusten, Tor Erik;Ryan, Kevin M.;Ryter, Stefan W.;Sabatini, David M.;Sadoshima, Junichi;Saha, Tapas;Saitoh, Tatsuya;Sakagami, Hiroshi;Sakai, Yasuyoshi;Salekdeh, Ghasem Hoseini;Salomoni, Paolo;Salvaterra, Paul M.;Salvesen, Guy;Salvioli, Rosa;Sanchez, Anthony M. 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M.;Simmons, Alison;Simon, Anna Katharina;Simon, Hans-Uwe;Simone, Cristiano;Simonsen, Anne;Sinclair, David A.;Singh, Rajat;Sinha, Debasish;Sinicrope, Frank A.;Sirko, Agnieszka;Siu, Parco M.;Sivridis, Efthimios;Skop, Vojtech;Skulachev, Vladimir P.;Slack, Ruth S.;Smaili, Soraya S.;Smith, Duncan R.;Soengas, Maria S.;Soldati, Thierry;Song, Xueqin;Sood, Anil K.;Soong, Tuck Wah;Sotgia, Federica;Spector, Stephen A.;Spies, Claudia D.;Springer, Wolfdieter;Srinivasula, Srinivasa M.;Stefanis, Leonidas;Steffan, Joan S.;Stendel, Ruediger;Stenmark, Harald;Stephanou, Anastasis;Stern, Stephan T.;Sternberg, Cinthya;Stork, Bjoern;Stralfors, Peter;Subauste, Carlos S.;Sui, Xinbing;Sulzer, David;Sun, Jiaren;Sun, Shi-Yong;Sun, Zhi-Jun;Sung, Joseph J. Y.;Suzuki, Kuninori;Suzuki, Toshihiko;Swanson, Michele S.;Swanton, Charles;Sweeney, Sean T.;Sy, Lai-King;Szabadkai, Gyorgy;Tabas, Ira;Taegtmeyer, Heinrich;Tafani, Marco;Takacs-Vellai, Krisztina;Takano, Yoshitaka;Takegawa, Kaoru;Takemura, Genzou;Takeshita, Fumihiko;Talbot, Nicholas J.;Tan, Kevin S. W.;Tanaka, Keiji;Tanaka, Kozo;Tang, Daolin;Tang, Dingzhong;Tanida, Isei;Tannous, Bakhos A.;Tavernarakis, Nektarios;Taylor, Graham S.;Taylor, Gregory A.;Taylor, J. 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通讯作者:
Klionsky, DJ
作者机构:
[Lukacs, Nicholas W.; Gestwicki, Jason E.; Lieberman, Andrew] Univ Michigan, Dept Pathol, Ann Arbor, MI 48109 USA.
[Kessel, David H.] Wayne State Univ, Sch Med, Dept Pharmacol, Detroit, MI 48201 USA.
[Into, Takeshi] Asahi Univ, Sch Dent, Dept Oral Microbiol, Div Oral Infect & Hlth Sci, Mizuho, Japan.
[Lotze, Michael T.; Tang, Daolin; Kang, Rui] Univ Pittsburgh, Inst Canc, Dept Surg, DAMP Lab,Hillman Canc Ctr, Pittsburgh, PA USA.
[Garcia-Manero, Guillermo] Univ Texas MD Anderson Canc Ctr, Dept Leukemia, Houston, TX 77030 USA.
通讯机构:
[Klionsky, Daniel J.] Univ Michigan, Inst Life Sci, Ann Arbor, MI 48109 USA.
语种:
英文
关键词:
autolysosome;autophagosome;flux;LC3;lysosome;phagophore;stress;vacuole
期刊:
AUTOPHAGY
ISSN:
1554-8627
年:
2012
卷:
8
期:
4
页码:
445-544
文献类别:
WOS:Review
所属学科:
ESI学科类别:分子生物学&遗传学;WOS学科类别:Cell Biology
入藏号:
基金类别:
National Institutes of Health Public Health Service [GM53396]
机构署名:
本校为其他机构
院系归属:
生命科学学院
摘要:
In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are 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 monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
参考文献:
Abeliovich H, 1999, EMBO J, V18, P6005, DOI 10.1093/emboj/18.21.6005
Abeliovich H, 2011, ANTIOXID REDOX SIGN, V14, P2003, DOI 10.1089/ars.2010.3807
Abeliovich H, 2009, AUTOPHAGY, V5, P925
AHLBERG J, 1985, J BIOL CHEM, V260, P5847
Aits S, 2009, INT J CANCER, V124, P1008, DOI 10.1002/ijc.24076

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