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

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 H Brumell, Ulf T BrunkWilfried Bursch, Nadine Camougrand, Eduardo Cebollero, Francesco Cecconi, Yingyu Chen, Lih-Shen Chin, Augustine Choi, Charleen T Chu, Jongkyeong Chung, Robert SB Clark, Peter GH Clarke, Steven G Clarke, Corinne Clave, John L Cleveland, Patrice Codogno, Maria I Colombo, Aba Coto-Montes, James M Cregg, Ana Maria Cuervo, Jayanta Debnath, Patrick B Dennis, Phillip A Dennis, Francesca Demarchi, Vojo Deretic, Rodney James Devenish, Frederica Di Sano, J Fred Dice, Clark W Distelhorst, Savithramma Dinesh-Kumar, N Tony Eissa, Marian DiFiglia, Mojgan Djavaheri-Mergny, Frank C Dorsey, Wulf Droge, Michel Dron, William A Dunn Jr, Michael Duszenko, Zvulun Elazar, Audrey Esclatine, Eeva-Liisa Eskelinen, Laszlo Fesus, Kim Finley, Jose M Fuentes, Juan Fueyo, Kozo Fujisaki, Brigitte Galliot, Fen-Biao Gao, David A Gewirtz, Spencer B Gibson, Antje Gohla, Alfred L Goldberg, Ramon Gonzalez, Cristina Gonzalez-Estevez, Sharon Gorski, Roberta A Gottlieb, Dieter Haussinger, You-Wen He, Kim Heidenreich, Joseph A Hill, Maria Hoyer-Hansen, Xun Hu, Wei-Pang Huang, Akiko Iwasaki, Marja Jaattela, William T Jackson, Xuejun Jiang, Shengkan Jin, Terje Johansen, Jae U Jung, Motoni Kadowaki, Chanhee Kang, Ameeta Kelekar, David H Kessel, Jan AKW Kiel, Adi Kimchi, Hong Pyo Kim, Timothy J Kinsella, Kirill Kiselyov, Katsuhiko Kitamoto, Erwin Knecht, Masaaki Komatsu, Eiki Kominami, Seiji Kondo, Attila L Kovacs, Guido Kroemer, Chia-Yi Kuan, Rakesh Kumar, Mondira Kundu, Jacques Landry, Marianne Laporte, Weidong Le, Huan-Yao Lei, Beth Levine, Andrew Lieberman, Kah-Leong Lim, Fu-Cheng Lin, Willisa Liou, Leroy F Liu, Gabriel Lopez-Berestein, Carlos Lopez-Otin, Bo Lu, Kay F MacLeod, Walter Malorni, Wim Martinet, Ken Matsuoka, Josef Mautner, Alfred J Meijer, Alicia Melendez, Paul Michels, Giovanni Miotto, Wilhelm P Mistiaen, Noboru Mizushima, Baharia Mogradi, Michael N Moore, Paula I Moreira, Yuji Moriyasu, Tomasz Motyl, Christian Munz, Leon O Murphy, Naweed Naqvi, Thomas P Neufeld, Ichizo Nishino, Ralph A Nixon, Takeshi Noda, Bernd Nurnberg, Michinaga Ogawa, Nancy L Oleinick, Laura J Olsen, Bulent Ozpolat, Shoshana Paglin, Glen E Palmer, Issidora Papassideri, Miles Parkes, David H Perlmutter, George Perry, Mauro Piacentini, Ronit Pinkas-Kramarski, Mark Prescott, Tassula Proikas-Cezanne, Nina Raben, Abdelhaq Rami, Fulvio Reggiori, Barbel Rohrer, David C Rubinsztein, Kevin M Ryan, Junichi Sadoshima, Hiroshi Sakagami, Yasuyoshi Sakai, Marco Sandri, Chihiro Sasakawa, Miklos Sass, Claudio Schneider, Per O Seglen, Oleksandr Seleverstov, Jeffrey Settleman, John J Shacka, Irving M Shapiro, Andrei Sibirny, Elaine CM Silva-Zacarin, Hans-Uwe Simon, Cristiano Simone, Anne Simonsen, Mark A Smith, Katharina Spanel-Borowski, Vickram Srinivas, Meredith Steeves, Harald Stenmark, Per E Stromhaug, Carlos S Subauste, Seiichiro Sugimoto, David Sulzer, Toshihiko Suzuki, Michele S Swanson, Ira Tabas, Fumihiko Takeshita, Nicholas J Talbot, Zsolt Talloczy, 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 L Uzcategui, Ida van der Klei, Eva C Vaquero, Tibor Vellai, Michael W Vogel, Hong-Gang Wang, Paul 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

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1779 Citations (Scopus)

Abstract

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 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.
Original languageEnglish
Pages (from-to)151 - 175
Number of pages24
JournalAutophagy
Volume4
Publication statusPublished - 2008

Cite this

Klionsky, D. J., Abeliovich, H., Agostinis, P., Agrawal, D. K., Aliev, G., Askew, D. S., ... Deter, R. L. (2008). Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy, 4, 151 - 175.
Klionsky, Daniel J ; Abeliovich, Hagai ; Agostinis, Patrizia ; Agrawal, Devendra K ; Aliev, Gjumrakch ; Askew, David S ; Baba, Misuzu ; Baehrecke, Eric H ; Bahr, Ben A ; Ballabio, Andrea ; Bamber, Bruce A ; Bassham, Diane C ; Bergamini, Ettore ; Bi, Xiaoning ; Biard-Piechaczyk, Martine ; Blum, Janice S ; Bredesen, Dale E ; Brodsky, Jeffrey L ; Brumell, John H ; Brunk, Ulf T ; Bursch, Wilfried ; Camougrand, Nadine ; Cebollero, Eduardo ; Cecconi, Francesco ; Chen, Yingyu ; Chin, Lih-Shen ; Choi, Augustine ; Chu, Charleen T ; Chung, Jongkyeong ; Clark, Robert SB ; Clarke, Peter GH ; Clarke, Steven G ; Clave, Corinne ; Cleveland, John L ; Codogno, Patrice ; Colombo, Maria I ; Coto-Montes, Aba ; Cregg, James M ; Cuervo, Ana Maria ; Debnath, Jayanta ; Dennis, Patrick B ; Dennis, Phillip A ; Demarchi, Francesca ; Deretic, Vojo ; Devenish, Rodney James ; Di Sano, Frederica ; Dice, J Fred ; Distelhorst, Clark W ; Dinesh-Kumar, Savithramma ; Eissa, N Tony ; DiFiglia, Marian ; Djavaheri-Mergny, Mojgan ; Dorsey, Frank C ; Droge, Wulf ; Dron, Michel ; Dunn Jr, William A ; Duszenko, Michael ; Elazar, Zvulun ; Esclatine, Audrey ; Eskelinen, Eeva-Liisa ; Fesus, Laszlo ; Finley, Kim ; Fuentes, Jose M ; Fueyo, Juan ; Fujisaki, Kozo ; Galliot, Brigitte ; Gao, Fen-Biao ; Gewirtz, David A ; Gibson, Spencer B ; Gohla, Antje ; Goldberg, Alfred L ; Gonzalez, Ramon ; Gonzalez-Estevez, Cristina ; Gorski, Sharon ; Gottlieb, Roberta A ; Haussinger, Dieter ; He, You-Wen ; Heidenreich, Kim ; Hill, Joseph A ; Hoyer-Hansen, Maria ; Hu, Xun ; Huang, Wei-Pang ; Iwasaki, Akiko ; Jaattela, Marja ; Jackson, William T ; Jiang, Xuejun ; Jin, Shengkan ; Johansen, Terje ; Jung, Jae U ; Kadowaki, Motoni ; Kang, Chanhee ; Kelekar, Ameeta ; Kessel, David H ; Kiel, Jan AKW ; Kimchi, Adi ; Kim, Hong Pyo ; Kinsella, Timothy J ; Kiselyov, Kirill ; Kitamoto, Katsuhiko ; Knecht, Erwin ; Komatsu, Masaaki ; Kominami, Eiki ; Kondo, Seiji ; Kovacs, Attila L ; Kroemer, Guido ; Kuan, Chia-Yi ; Kumar, Rakesh ; Kundu, Mondira ; Landry, Jacques ; Laporte, Marianne ; Le, Weidong ; Lei, Huan-Yao ; Levine, Beth ; Lieberman, Andrew ; Lim, Kah-Leong ; Lin, Fu-Cheng ; Liou, Willisa ; Liu, Leroy F ; Lopez-Berestein, Gabriel ; Lopez-Otin, Carlos ; Lu, Bo ; MacLeod, Kay F ; Malorni, Walter ; Martinet, Wim ; Matsuoka, Ken ; Mautner, Josef ; Meijer, Alfred J ; Melendez, Alicia ; Michels, Paul ; Miotto, Giovanni ; Mistiaen, Wilhelm P ; Mizushima, Noboru ; Mogradi, Baharia ; Moore, Michael N ; Moreira, Paula I ; Moriyasu, Yuji ; Motyl, Tomasz ; Munz, Christian ; Murphy, Leon O ; Naqvi, Naweed ; Neufeld, Thomas P ; Nishino, Ichizo ; Nixon, Ralph A ; Noda, Takeshi ; Nurnberg, Bernd ; Ogawa, Michinaga ; Oleinick, Nancy L ; Olsen, Laura J ; Ozpolat, Bulent ; Paglin, Shoshana ; Palmer, Glen E ; Papassideri, Issidora ; Parkes, Miles ; Perlmutter, David H ; Perry, George ; Piacentini, Mauro ; Pinkas-Kramarski, Ronit ; Prescott, Mark ; Proikas-Cezanne, Tassula ; Raben, Nina ; Rami, Abdelhaq ; Reggiori, Fulvio ; Rohrer, Barbel ; Rubinsztein, David C ; Ryan, Kevin M ; Sadoshima, Junichi ; Sakagami, Hiroshi ; Sakai, Yasuyoshi ; Sandri, Marco ; Sasakawa, Chihiro ; Sass, Miklos ; Schneider, Claudio ; Seglen, Per O ; Seleverstov, Oleksandr ; Settleman, Jeffrey ; Shacka, John J ; Shapiro, Irving M ; Sibirny, Andrei ; Silva-Zacarin, Elaine CM ; Simon, Hans-Uwe ; Simone, Cristiano ; Simonsen, Anne ; Smith, Mark A ; Spanel-Borowski, Katharina ; Srinivas, Vickram ; Steeves, Meredith ; Stenmark, Harald ; Stromhaug, Per E ; Subauste, Carlos S ; Sugimoto, Seiichiro ; Sulzer, David ; Suzuki, Toshihiko ; Swanson, Michele S ; Tabas, Ira ; Takeshita, Fumihiko ; Talbot, Nicholas J ; Talloczy, Zsolt ; Tanaka, Keiji ; Tanaka, Kozo ; Tanida, Isei ; Taylor, Graham S ; Taylor, J Paul ; Terman, Alexei ; Tettamanti, Gianluca ; Thompson, Craig B ; Thumm, Michael ; Tolkovsky, Aviva M ; Tooze, Sharon A ; Truant, Ray ; Tumanovska, Lesya V ; Uchiyama, Yasuo ; Ueno, Takashi ; Uzcategui, Nestor L ; van der Klei, Ida ; Vaquero, Eva C ; Vellai, Tibor ; Vogel, Michael W ; Wang, Hong-Gang ; Webster, Paul ; Xi, Zhijun ; Xiao, Gutian ; Yahalom, Joachim ; Yang, Jin-Ming ; Yap, George ; Yin, Xiao-Ming ; Yoshimori, Tamotsu ; Yue, Zhenyu ; Yuzaki, Michisuke ; Zabirnyk, Olga ; Zheng, Xiaoxiang ; Zhu, Xiongwei ; Deter, Russell L. / Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. In: Autophagy. 2008 ; Vol. 4. pp. 151 - 175.
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title = "Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes",
abstract = "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 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.",
author = "Klionsky, {Daniel J} and Hagai Abeliovich and Patrizia Agostinis and Agrawal, {Devendra K} and Gjumrakch Aliev and Askew, {David S} and Misuzu Baba and Baehrecke, {Eric H} and Bahr, {Ben A} and Andrea Ballabio and Bamber, {Bruce A} and Bassham, {Diane C} and Ettore Bergamini and Xiaoning Bi and Martine Biard-Piechaczyk and Blum, {Janice S} and Bredesen, {Dale E} and Brodsky, {Jeffrey L} and Brumell, {John H} and Brunk, {Ulf T} and Wilfried Bursch and Nadine Camougrand and Eduardo Cebollero and Francesco Cecconi and Yingyu Chen and Lih-Shen Chin and Augustine Choi and Chu, {Charleen T} and Jongkyeong Chung and Clark, {Robert SB} and Clarke, {Peter GH} and Clarke, {Steven G} and Corinne Clave and Cleveland, {John L} and Patrice Codogno and Colombo, {Maria I} and Aba Coto-Montes and Cregg, {James M} and Cuervo, {Ana Maria} and Jayanta Debnath and Dennis, {Patrick B} and Dennis, {Phillip A} and Francesca Demarchi and Vojo Deretic and Devenish, {Rodney James} and {Di Sano}, Frederica and Dice, {J Fred} and Distelhorst, {Clark W} and Savithramma Dinesh-Kumar and Eissa, {N Tony} and Marian DiFiglia and Mojgan Djavaheri-Mergny and Dorsey, {Frank C} and Wulf Droge and Michel Dron and {Dunn Jr}, {William A} and Michael Duszenko and Zvulun Elazar and Audrey Esclatine and Eeva-Liisa Eskelinen and Laszlo Fesus and Kim Finley and Fuentes, {Jose M} and Juan Fueyo and Kozo Fujisaki and Brigitte Galliot and Fen-Biao Gao and Gewirtz, {David A} and Gibson, {Spencer B} and Antje Gohla and Goldberg, {Alfred L} and Ramon Gonzalez and Cristina Gonzalez-Estevez and Sharon Gorski and Gottlieb, {Roberta A} and Dieter Haussinger and You-Wen He and Kim Heidenreich and Hill, {Joseph A} and Maria Hoyer-Hansen and Xun Hu and Wei-Pang Huang and Akiko Iwasaki and Marja Jaattela and Jackson, {William T} and Xuejun Jiang and Shengkan Jin and Terje Johansen and Jung, {Jae U} and Motoni Kadowaki and Chanhee Kang and Ameeta Kelekar and Kessel, {David H} and Kiel, {Jan AKW} and Adi Kimchi and Kim, {Hong Pyo} and Kinsella, {Timothy J} and Kirill Kiselyov and Katsuhiko Kitamoto and Erwin Knecht and Masaaki Komatsu and Eiki Kominami and Seiji Kondo and Kovacs, {Attila L} and Guido Kroemer and Chia-Yi Kuan and Rakesh Kumar and Mondira Kundu and Jacques Landry and Marianne Laporte and Weidong Le and Huan-Yao Lei and Beth Levine and Andrew Lieberman and Kah-Leong Lim and Fu-Cheng Lin and Willisa Liou and Liu, {Leroy F} and Gabriel Lopez-Berestein and Carlos Lopez-Otin and Bo Lu and MacLeod, {Kay F} and Walter Malorni and Wim Martinet and Ken Matsuoka and Josef Mautner and Meijer, {Alfred J} and Alicia Melendez and Paul Michels and Giovanni Miotto and Mistiaen, {Wilhelm P} and Noboru Mizushima and Baharia Mogradi and Moore, {Michael N} and Moreira, {Paula I} and Yuji Moriyasu and Tomasz Motyl and Christian Munz and Murphy, {Leon O} and Naweed Naqvi and Neufeld, {Thomas P} and Ichizo Nishino and Nixon, {Ralph A} and Takeshi Noda and Bernd Nurnberg and Michinaga Ogawa and Oleinick, {Nancy L} and Olsen, {Laura J} and Bulent Ozpolat and Shoshana Paglin and Palmer, {Glen E} and Issidora Papassideri and Miles Parkes and Perlmutter, {David H} and George Perry and Mauro Piacentini and Ronit Pinkas-Kramarski and Mark Prescott and Tassula Proikas-Cezanne and Nina Raben and Abdelhaq Rami and Fulvio Reggiori and Barbel Rohrer and Rubinsztein, {David C} and Ryan, {Kevin M} and Junichi Sadoshima and Hiroshi Sakagami and Yasuyoshi Sakai and Marco Sandri and Chihiro Sasakawa and Miklos Sass and Claudio Schneider and Seglen, {Per O} and Oleksandr Seleverstov and Jeffrey Settleman and Shacka, {John J} and Shapiro, {Irving M} and Andrei Sibirny and Silva-Zacarin, {Elaine CM} and Hans-Uwe Simon and Cristiano Simone and Anne Simonsen and Smith, {Mark A} and Katharina Spanel-Borowski and Vickram Srinivas and Meredith Steeves and Harald Stenmark and Stromhaug, {Per E} and Subauste, {Carlos S} and Seiichiro Sugimoto and David Sulzer and Toshihiko Suzuki and Swanson, {Michele S} and Ira Tabas and Fumihiko Takeshita and Talbot, {Nicholas J} and Zsolt Talloczy and Keiji Tanaka and Kozo Tanaka and Isei Tanida and Taylor, {Graham S} and Taylor, {J Paul} and Alexei Terman and Gianluca Tettamanti and Thompson, {Craig B} and Michael Thumm and Tolkovsky, {Aviva M} and Tooze, {Sharon A} and Ray Truant and Tumanovska, {Lesya V} and Yasuo Uchiyama and Takashi Ueno and Uzcategui, {Nestor L} and {van der Klei}, Ida and Vaquero, {Eva C} and Tibor Vellai and Vogel, {Michael W} and Hong-Gang Wang and Paul Webster and Zhijun Xi and Gutian Xiao and Joachim Yahalom and Jin-Ming Yang and George Yap and Xiao-Ming Yin and Tamotsu Yoshimori and Zhenyu Yue and Michisuke Yuzaki and Olga Zabirnyk and Xiaoxiang Zheng and Xiongwei Zhu and Deter, {Russell L}",
year = "2008",
language = "English",
volume = "4",
pages = "151 -- 175",
journal = "Autophagy",
issn = "1554-8627",
publisher = "Taylor & Francis",

}

Klionsky, DJ, Abeliovich, H, Agostinis, P, Agrawal, DK, Aliev, G, Askew, DS, Baba, M, Baehrecke, EH, Bahr, BA, Ballabio, A, Bamber, BA, Bassham, DC, Bergamini, E, Bi, X, Biard-Piechaczyk, M, Blum, JS, Bredesen, DE, Brodsky, JL, Brumell, JH, Brunk, UT, Bursch, W, Camougrand, N, Cebollero, E, Cecconi, F, Chen, Y, Chin, L-S, Choi, A, Chu, CT, Chung, J, Clark, RSB, Clarke, PGH, Clarke, SG, Clave, C, Cleveland, JL, Codogno, P, Colombo, MI, Coto-Montes, A, Cregg, JM, Cuervo, AM, Debnath, J, Dennis, PB, Dennis, PA, Demarchi, F, Deretic, V, Devenish, RJ, Di Sano, F, Dice, JF, Distelhorst, CW, Dinesh-Kumar, S, Eissa, NT, DiFiglia, M, Djavaheri-Mergny, M, Dorsey, FC, Droge, W, Dron, M, Dunn Jr, WA, Duszenko, M, Elazar, Z, Esclatine, A, Eskelinen, E-L, Fesus, L, Finley, K, Fuentes, JM, Fueyo, J, Fujisaki, K, Galliot, B, Gao, F-B, Gewirtz, DA, Gibson, SB, Gohla, A, Goldberg, AL, Gonzalez, R, Gonzalez-Estevez, C, Gorski, S, Gottlieb, RA, Haussinger, D, He, Y-W, Heidenreich, K, Hill, JA, Hoyer-Hansen, M, Hu, X, Huang, W-P, Iwasaki, A, Jaattela, M, Jackson, WT, Jiang, X, Jin, S, Johansen, T, Jung, JU, Kadowaki, M, Kang, C, Kelekar, A, Kessel, DH, Kiel, JAKW, Kimchi, A, Kim, HP, Kinsella, TJ, Kiselyov, K, Kitamoto, K, Knecht, E, Komatsu, M, Kominami, E, Kondo, S, Kovacs, AL, Kroemer, G, Kuan, C-Y, Kumar, R, Kundu, M, Landry, J, Laporte, M, Le, W, Lei, H-Y, Levine, B, Lieberman, A, Lim, K-L, Lin, F-C, Liou, W, Liu, LF, Lopez-Berestein, G, Lopez-Otin, C, Lu, B, MacLeod, KF, Malorni, W, Martinet, W, Matsuoka, K, Mautner, J, Meijer, AJ, Melendez, A, Michels, P, Miotto, G, Mistiaen, WP, Mizushima, N, Mogradi, B, Moore, MN, Moreira, PI, Moriyasu, Y, Motyl, T, Munz, C, Murphy, LO, Naqvi, N, Neufeld, TP, Nishino, I, Nixon, RA, Noda, T, Nurnberg, B, Ogawa, M, Oleinick, NL, Olsen, LJ, Ozpolat, B, Paglin, S, Palmer, GE, Papassideri, I, Parkes, M, Perlmutter, DH, Perry, G, Piacentini, M, Pinkas-Kramarski, R, Prescott, M, Proikas-Cezanne, T, Raben, N, Rami, A, Reggiori, F, Rohrer, B, Rubinsztein, DC, Ryan, KM, Sadoshima, J, Sakagami, H, Sakai, Y, Sandri, M, Sasakawa, C, Sass, M, Schneider, C, Seglen, PO, Seleverstov, O, Settleman, J, Shacka, JJ, Shapiro, IM, Sibirny, A, Silva-Zacarin, ECM, Simon, H-U, Simone, C, Simonsen, A, Smith, MA, Spanel-Borowski, K, Srinivas, V, Steeves, M, Stenmark, H, Stromhaug, PE, Subauste, CS, Sugimoto, S, Sulzer, D, Suzuki, T, Swanson, MS, Tabas, I, Takeshita, F, Talbot, NJ, Talloczy, Z, Tanaka, K, Tanaka, K, Tanida, I, Taylor, GS, Taylor, JP, Terman, A, Tettamanti, G, Thompson, CB, Thumm, M, Tolkovsky, AM, Tooze, SA, Truant, R, Tumanovska, LV, Uchiyama, Y, Ueno, T, Uzcategui, NL, van der Klei, I, Vaquero, EC, Vellai, T, Vogel, MW, Wang, H-G, Webster, P, Xi, Z, Xiao, G, Yahalom, J, Yang, J-M, Yap, G, Yin, X-M, Yoshimori, T, Yue, Z, Yuzaki, M, Zabirnyk, O, Zheng, X, Zhu, X & Deter, RL 2008, 'Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes', Autophagy, vol. 4, pp. 151 - 175.

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

In: Autophagy, Vol. 4, 2008, p. 151 - 175.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes

AU - Klionsky, Daniel J

AU - Abeliovich, Hagai

AU - Agostinis, Patrizia

AU - Agrawal, Devendra K

AU - Aliev, Gjumrakch

AU - Askew, David S

AU - Baba, Misuzu

AU - Baehrecke, Eric H

AU - Bahr, Ben A

AU - Ballabio, Andrea

AU - Bamber, Bruce A

AU - Bassham, Diane C

AU - Bergamini, Ettore

AU - Bi, Xiaoning

AU - Biard-Piechaczyk, Martine

AU - Blum, Janice S

AU - Bredesen, Dale E

AU - Brodsky, Jeffrey L

AU - Brumell, John H

AU - Brunk, Ulf T

AU - Bursch, Wilfried

AU - Camougrand, Nadine

AU - Cebollero, Eduardo

AU - Cecconi, Francesco

AU - Chen, Yingyu

AU - Chin, Lih-Shen

AU - Choi, Augustine

AU - Chu, Charleen T

AU - Chung, Jongkyeong

AU - Clark, Robert SB

AU - Clarke, Peter GH

AU - Clarke, Steven G

AU - Clave, Corinne

AU - Cleveland, John L

AU - Codogno, Patrice

AU - Colombo, Maria I

AU - Coto-Montes, Aba

AU - Cregg, James M

AU - Cuervo, Ana Maria

AU - Debnath, Jayanta

AU - Dennis, Patrick B

AU - Dennis, Phillip A

AU - Demarchi, Francesca

AU - Deretic, Vojo

AU - Devenish, Rodney James

AU - Di Sano, Frederica

AU - Dice, J Fred

AU - Distelhorst, Clark W

AU - Dinesh-Kumar, Savithramma

AU - Eissa, N Tony

AU - DiFiglia, Marian

AU - Djavaheri-Mergny, Mojgan

AU - Dorsey, Frank C

AU - Droge, Wulf

AU - Dron, Michel

AU - Dunn Jr, William A

AU - Duszenko, Michael

AU - Elazar, Zvulun

AU - Esclatine, Audrey

AU - Eskelinen, Eeva-Liisa

AU - Fesus, Laszlo

AU - Finley, Kim

AU - Fuentes, Jose M

AU - Fueyo, Juan

AU - Fujisaki, Kozo

AU - Galliot, Brigitte

AU - Gao, Fen-Biao

AU - Gewirtz, David A

AU - Gibson, Spencer B

AU - Gohla, Antje

AU - Goldberg, Alfred L

AU - Gonzalez, Ramon

AU - Gonzalez-Estevez, Cristina

AU - Gorski, Sharon

AU - Gottlieb, Roberta A

AU - Haussinger, Dieter

AU - He, You-Wen

AU - Heidenreich, Kim

AU - Hill, Joseph A

AU - Hoyer-Hansen, Maria

AU - Hu, Xun

AU - Huang, Wei-Pang

AU - Iwasaki, Akiko

AU - Jaattela, Marja

AU - Jackson, William T

AU - Jiang, Xuejun

AU - Jin, Shengkan

AU - Johansen, Terje

AU - Jung, Jae U

AU - Kadowaki, Motoni

AU - Kang, Chanhee

AU - Kelekar, Ameeta

AU - Kessel, David H

AU - Kiel, Jan AKW

AU - Kimchi, Adi

AU - Kim, Hong Pyo

AU - Kinsella, Timothy J

AU - Kiselyov, Kirill

AU - Kitamoto, Katsuhiko

AU - Knecht, Erwin

AU - Komatsu, Masaaki

AU - Kominami, Eiki

AU - Kondo, Seiji

AU - Kovacs, Attila L

AU - Kroemer, Guido

AU - Kuan, Chia-Yi

AU - Kumar, Rakesh

AU - Kundu, Mondira

AU - Landry, Jacques

AU - Laporte, Marianne

AU - Le, Weidong

AU - Lei, Huan-Yao

AU - Levine, Beth

AU - Lieberman, Andrew

AU - Lim, Kah-Leong

AU - Lin, Fu-Cheng

AU - Liou, Willisa

AU - Liu, Leroy F

AU - Lopez-Berestein, Gabriel

AU - Lopez-Otin, Carlos

AU - Lu, Bo

AU - MacLeod, Kay F

AU - Malorni, Walter

AU - Martinet, Wim

AU - Matsuoka, Ken

AU - Mautner, Josef

AU - Meijer, Alfred J

AU - Melendez, Alicia

AU - Michels, Paul

AU - Miotto, Giovanni

AU - Mistiaen, Wilhelm P

AU - Mizushima, Noboru

AU - Mogradi, Baharia

AU - Moore, Michael N

AU - Moreira, Paula I

AU - Moriyasu, Yuji

AU - Motyl, Tomasz

AU - Munz, Christian

AU - Murphy, Leon O

AU - Naqvi, Naweed

AU - Neufeld, Thomas P

AU - Nishino, Ichizo

AU - Nixon, Ralph A

AU - Noda, Takeshi

AU - Nurnberg, Bernd

AU - Ogawa, Michinaga

AU - Oleinick, Nancy L

AU - Olsen, Laura J

AU - Ozpolat, Bulent

AU - Paglin, Shoshana

AU - Palmer, Glen E

AU - Papassideri, Issidora

AU - Parkes, Miles

AU - Perlmutter, David H

AU - Perry, George

AU - Piacentini, Mauro

AU - Pinkas-Kramarski, Ronit

AU - Prescott, Mark

AU - Proikas-Cezanne, Tassula

AU - Raben, Nina

AU - Rami, Abdelhaq

AU - Reggiori, Fulvio

AU - Rohrer, Barbel

AU - Rubinsztein, David C

AU - Ryan, Kevin M

AU - Sadoshima, Junichi

AU - Sakagami, Hiroshi

AU - Sakai, Yasuyoshi

AU - Sandri, Marco

AU - Sasakawa, Chihiro

AU - Sass, Miklos

AU - Schneider, Claudio

AU - Seglen, Per O

AU - Seleverstov, Oleksandr

AU - Settleman, Jeffrey

AU - Shacka, John J

AU - Shapiro, Irving M

AU - Sibirny, Andrei

AU - Silva-Zacarin, Elaine CM

AU - Simon, Hans-Uwe

AU - Simone, Cristiano

AU - Simonsen, Anne

AU - Smith, Mark A

AU - Spanel-Borowski, Katharina

AU - Srinivas, Vickram

AU - Steeves, Meredith

AU - Stenmark, Harald

AU - Stromhaug, Per E

AU - Subauste, Carlos S

AU - Sugimoto, Seiichiro

AU - Sulzer, David

AU - Suzuki, Toshihiko

AU - Swanson, Michele S

AU - Tabas, Ira

AU - Takeshita, Fumihiko

AU - Talbot, Nicholas J

AU - Talloczy, Zsolt

AU - Tanaka, Keiji

AU - Tanaka, Kozo

AU - Tanida, Isei

AU - Taylor, Graham S

AU - Taylor, J Paul

AU - Terman, Alexei

AU - Tettamanti, Gianluca

AU - Thompson, Craig B

AU - Thumm, Michael

AU - Tolkovsky, Aviva M

AU - Tooze, Sharon A

AU - Truant, Ray

AU - Tumanovska, Lesya V

AU - Uchiyama, Yasuo

AU - Ueno, Takashi

AU - Uzcategui, Nestor L

AU - van der Klei, Ida

AU - Vaquero, Eva C

AU - Vellai, Tibor

AU - Vogel, Michael W

AU - Wang, Hong-Gang

AU - Webster, Paul

AU - Xi, Zhijun

AU - Xiao, Gutian

AU - Yahalom, Joachim

AU - Yang, Jin-Ming

AU - Yap, George

AU - Yin, Xiao-Ming

AU - Yoshimori, Tamotsu

AU - Yue, Zhenyu

AU - Yuzaki, Michisuke

AU - Zabirnyk, Olga

AU - Zheng, Xiaoxiang

AU - Zhu, Xiongwei

AU - Deter, Russell L

PY - 2008

Y1 - 2008

N2 - 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 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.

AB - 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 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.

UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18188003

M3 - Article

VL - 4

SP - 151

EP - 175

JO - Autophagy

JF - Autophagy

SN - 1554-8627

ER -

Klionsky DJ, Abeliovich H, Agostinis P, Agrawal DK, Aliev G, Askew DS et al. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy. 2008;4:151 - 175.