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 Blum, Dale E. Bredesen, Jeffrey L. Brodsky, John H. Brumell, Ulf T. Brunk & 216 others Wilfried Bursch, Nadine Camougrand, Eduardo Cebollero, Francesco Cecconi, Yingyu Chen, Lih Shen Chin, Augustine Choi, Charleen T. Chu, Jongkyeong Chung, Peter G H Clarke, Robert S B Clark, Steven G. Clarke, Corinne Clavé, John L. Cleveland, Patrice Codogno, María I. Colombo, Ana Cotomontes, James M. Cregg, Ana Maria Cuervo, Jayanta Debnath, Francesca Demarchi, Patrick B. Dennis, Phillip A. Dennis, Vojo Deretic, Rodney J. Devenish, Federica Di Sano, J. Fred Dice, Marian DiFiglia, Savithramma Dinesh-Kumar, Clark W. Distelhorst, Mojgan Djavaheri-Mergny, Frank C. Dorsey, Wulf Dröge, Michel Dron, William A. Dunn, Michael Duszenko, N. Tony Eissa, Zvulun Elazar, Audrey Esclatine, Eeva Liisa Eskelinen, László Fésüs, Kim D. Finley, José 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 González-Estévez, Sharon 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 Jin, Terje Johansen, Jae U. Jung, Motoni Kadowaki, Chanhee Kang, Ameeta Kelekar, David H. Kessel, Jan A K W Kiel, Pyo Kim Hong, Adi Kimchi, Timothy J. Kinsella, Kirill 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 Yao Lei, Michael J. Lenardo, Beth Levine, Andrew Lieberman, Kah Leong Lim, Fu Cheng Lin, Willisa Liou, Leroy F. Liu, Gabriel Lopez-Berestein, Carlos López-Otín, Bo Lu, Kay F. Macleod, Walter Malorni, Wim Martinet, Ken Matsuoka, Josef Mautner, Alfred J. Meijer, Alicia Meléndez, Paul Michels, Giovanni Miotto, Wilhelm P. Mistiaen, Noboru Mizushima, Baharia Mograbi, Iryna Monastyrska, Michael N. Moore, Paula I. Moreira, Yuji Moriyasu, Tomasz 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, Bulent Ozpolat, Shoshana Paglin, Glen E. Palmer, Issidora Papassideri, Miles Parkes, David H. Perlmutter, George Perry, Mauro Piacentini, Ronit Pinkas-Kramarski, Mark Prescott, Tassula Proikascezanne, Nina Raben, Abdelhaq Rami, Fulvio Reggiori, Bärbel 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 C M Silva-Zacarin, Hans Uwe Simon, Crisfiano 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 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, Néstor L. Uzcátegui, Ida Van Der Klei, Eva C. Vaquero, Tibor Vellai, Michael W. Vogel, Hong Gang Wang, Paul Webster, John W. Wiley, Zhijun Xi, Gutian Xiao, Joachim Yahalom, Jin Ming Yang, George Yap, Xiao-Ming Yin, Tamotsu Yoshimori, Li Yu, Zhenyu Yue, Michisuke Yuzaki, Olga Zabirnyk, Xiaoxiang Zheng, Xiongwei Zhu, Russell L. Deter

Research output: Contribution to journalArticle

1745 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 pages25
JournalAutophagy
Volume4
Issue number2
StatePublished - Feb 16 2008

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Autophagy
Eukaryota
Guidelines
Vacuoles
Lysosomes
Fungi
Yeasts
Research Personnel
Research

Keywords

  • Autolysosome
  • Autophagosome
  • Flux
  • Lysosome
  • Phagophore
  • Stress
  • Vacuole

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

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(2), 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; 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; Clarke, Peter G H; Clark, Robert S B; Clarke, Steven G.; Clavé, Corinne; Cleveland, John L.; Codogno, Patrice; Colombo, María I.; Cotomontes, Ana; Cregg, James M.; Cuervo, Ana Maria; Debnath, Jayanta; Demarchi, Francesca; Dennis, Patrick B.; Dennis, Phillip A.; Deretic, Vojo; Devenish, Rodney J.; Di Sano, Federica; Dice, J. Fred; DiFiglia, Marian; Dinesh-Kumar, Savithramma; Distelhorst, Clark W.; Djavaheri-Mergny, Mojgan; Dorsey, Frank C.; Dröge, Wulf; Dron, Michel; Dunn, William A.; Duszenko, Michael; Eissa, N. Tony; Elazar, Zvulun; Esclatine, Audrey; Eskelinen, Eeva Liisa; Fésüs, László; Finley, Kim D.; Fuentes, José M.; Fueyo, Juan; Fujisaki, Kozo; Galliot, Brigitte; Gao, Fen Biao; Gewirtz, David A.; Gibson, Spencer B.; Gohla, Antje; Goldberg, Alfred L.; Gonzalez, Ramon; González-Estévez, Cristina; Gorski, Sharon; Gottlieb, Roberta A.; Häussinger, Dieter; He, You Wen; Heidenreich, Kim; Hill, Joseph A.; Høyer-Hansen, Maria; Hu, Xun; Huang, Wei Pang; Iwasaki, Akiko; Jäättelä, Marja; Jackson, William T.; Jiang, Xuejun; Jin, Shengkan; Johansen, Terje; Jung, Jae U.; Kadowaki, Motoni; Kang, Chanhee; Kelekar, Ameeta; Kessel, David H.; Kiel, Jan A K W; Hong, Pyo Kim; Kimchi, Adi; Kinsella, Timothy J.; Kiselyov, Kirill; Kitamoto, Katsuhiko; Knecht, Erwin; Komatsu, Masaaki; Kominami, Eiki; Kondo, Seiji; Kovács, Attila L.; Kroemer, Guido; Kuan, Chia Yi; Kumar, Rakesh; Kundu, Mondira; Landry, Jacques; Laporte, Marianne; Le, Weidong; Lei, Huan Yao; Lenardo, Michael J.; Levine, Beth; Lieberman, Andrew; Lim, Kah Leong; Lin, Fu Cheng; Liou, Willisa; Liu, Leroy F.; Lopez-Berestein, Gabriel; López-Otín, Carlos; Lu, Bo; Macleod, Kay F.; Malorni, Walter; Martinet, Wim; Matsuoka, Ken; Mautner, Josef; Meijer, Alfred J.; Meléndez, Alicia; Michels, Paul; Miotto, Giovanni; Mistiaen, Wilhelm P.; Mizushima, Noboru; Mograbi, Baharia; Monastyrska, Iryna; Moore, Michael N.; Moreira, Paula I.; Moriyasu, Yuji; Motyl, Tomasz; Münz, Christian; Murphy, Leon O.; Naqvi, Naweed I.; Neufeld, Thomas P.; Nishino, Ichizo; Nixon, Ralph A.; Noda, Takeshi; Nürnberg, 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; Proikascezanne, Tassula; Raben, Nina; Rami, Abdelhaq; Reggiori, Fulvio; Rohrer, Bärbel; 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 C M; Simon, Hans Uwe; Simone, Crisfiano; 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.; Tallóczy, 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; Uzcátegui, Néstor L.; Van Der Klei, Ida; Vaquero, Eva C.; Vellai, Tibor; Vogel, Michael W.; Wang, Hong Gang; Webster, Paul; Wiley, John W.; Xi, Zhijun; Xiao, Gutian; Yahalom, Joachim; Yang, Jin Ming; Yap, George; Yin, Xiao-Ming; Yoshimori, Tamotsu; Yu, Li; Yue, Zhenyu; Yuzaki, Michisuke; Zabirnyk, Olga; Zheng, Xiaoxiang; Zhu, Xiongwei; Deter, Russell L.

In: Autophagy, Vol. 4, No. 2, 16.02.2008, p. 151-175.

Research output: Contribution to journalArticle

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, J, Bredesen, DE, Brodsky, JL, Brumell, JH, Brunk, UT, Bursch, W, Camougrand, N, Cebollero, E, Cecconi, F, Chen, Y, Chin, LS, Choi, A, Chu, CT, Chung, J, Clarke, PGH, Clark, RSB, Clarke, SG, Clavé, C, Cleveland, JL, Codogno, P, Colombo, MI, Cotomontes, A, Cregg, JM, Cuervo, AM, Debnath, J, Demarchi, F, Dennis, PB, Dennis, PA, Deretic, V, Devenish, RJ, Di Sano, F, Dice, JF, DiFiglia, M, Dinesh-Kumar, S, Distelhorst, CW, Djavaheri-Mergny, M, Dorsey, FC, Dröge, W, Dron, M, Dunn, WA, Duszenko, M, Eissa, NT, Elazar, Z, Esclatine, A, Eskelinen, EL, Fésüs, L, Finley, KD, Fuentes, JM, Fueyo, J, Fujisaki, K, Galliot, B, Gao, FB, Gewirtz, DA, Gibson, SB, Gohla, A, Goldberg, AL, Gonzalez, R, González-Estévez, C, Gorski, S, Gottlieb, RA, Häussinger, D, He, YW, Heidenreich, K, Hill, JA, Høyer-Hansen, M, Hu, X, Huang, WP, Iwasaki, A, Jäättelä, M, Jackson, WT, Jiang, X, Jin, S, Johansen, T, Jung, JU, Kadowaki, M, Kang, C, Kelekar, A, Kessel, DH, Kiel, JAKW, Hong, PK, Kimchi, A, Kinsella, TJ, Kiselyov, K, Kitamoto, K, Knecht, E, Komatsu, M, Kominami, E, Kondo, S, Kovács, AL, Kroemer, G, Kuan, CY, Kumar, R, Kundu, M, Landry, J, Laporte, M, Le, W, Lei, HY, Lenardo, MJ, Levine, B, Lieberman, A, Lim, KL, Lin, FC, Liou, W, Liu, LF, Lopez-Berestein, G, López-Otín, C, Lu, B, Macleod, KF, Malorni, W, Martinet, W, Matsuoka, K, Mautner, J, Meijer, AJ, Meléndez, A, Michels, P, Miotto, G, Mistiaen, WP, Mizushima, N, Mograbi, B, Monastyrska, I, Moore, MN, Moreira, PI, Moriyasu, Y, Motyl, T, Münz, C, Murphy, LO, Naqvi, NI, Neufeld, TP, Nishino, I, Nixon, RA, Noda, T, Nürnberg, 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, Proikascezanne, 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, HU, 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, Tallóczy, 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, Uzcátegui, NL, Van Der Klei, I, Vaquero, EC, Vellai, T, Vogel, MW, Wang, HG, Webster, P, Wiley, JW, Xi, Z, Xiao, G, Yahalom, J, Yang, JM, Yap, G, Yin, X-M, Yoshimori, T, Yu, L, 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, no. 2, pp. 151-175.
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 Feb 16;4(2):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 ; 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 ; Clarke, Peter G H ; Clark, Robert S B ; Clarke, Steven G. ; Clavé, Corinne ; Cleveland, John L. ; Codogno, Patrice ; Colombo, María I. ; Cotomontes, Ana ; Cregg, James M. ; Cuervo, Ana Maria ; Debnath, Jayanta ; Demarchi, Francesca ; Dennis, Patrick B. ; Dennis, Phillip A. ; Deretic, Vojo ; Devenish, Rodney J. ; Di Sano, Federica ; Dice, J. Fred ; DiFiglia, Marian ; Dinesh-Kumar, Savithramma ; Distelhorst, Clark W. ; Djavaheri-Mergny, Mojgan ; Dorsey, Frank C. ; Dröge, Wulf ; Dron, Michel ; Dunn, William A. ; Duszenko, Michael ; Eissa, N. Tony ; Elazar, Zvulun ; Esclatine, Audrey ; Eskelinen, Eeva Liisa ; Fésüs, László ; Finley, Kim D. ; Fuentes, José M. ; Fueyo, Juan ; Fujisaki, Kozo ; Galliot, Brigitte ; Gao, Fen Biao ; Gewirtz, David A. ; Gibson, Spencer B. ; Gohla, Antje ; Goldberg, Alfred L. ; Gonzalez, Ramon ; González-Estévez, Cristina ; Gorski, Sharon ; Gottlieb, Roberta A. ; Häussinger, Dieter ; He, You Wen ; Heidenreich, Kim ; Hill, Joseph A. ; Høyer-Hansen, Maria ; Hu, Xun ; Huang, Wei Pang ; Iwasaki, Akiko ; Jäättelä, Marja ; Jackson, William T. ; Jiang, Xuejun ; Jin, Shengkan ; Johansen, Terje ; Jung, Jae U. ; Kadowaki, Motoni ; Kang, Chanhee ; Kelekar, Ameeta ; Kessel, David H. ; Kiel, Jan A K W ; Hong, Pyo Kim ; Kimchi, Adi ; Kinsella, Timothy J. ; Kiselyov, Kirill ; Kitamoto, Katsuhiko ; Knecht, Erwin ; Komatsu, Masaaki ; Kominami, Eiki ; Kondo, Seiji ; Kovács, Attila L. ; Kroemer, Guido ; Kuan, Chia Yi ; Kumar, Rakesh ; Kundu, Mondira ; Landry, Jacques ; Laporte, Marianne ; Le, Weidong ; Lei, Huan Yao ; Lenardo, Michael J. ; Levine, Beth ; Lieberman, Andrew ; Lim, Kah Leong ; Lin, Fu Cheng ; Liou, Willisa ; Liu, Leroy F. ; Lopez-Berestein, Gabriel ; López-Otín, Carlos ; Lu, Bo ; Macleod, Kay F. ; Malorni, Walter ; Martinet, Wim ; Matsuoka, Ken ; Mautner, Josef ; Meijer, Alfred J. ; Meléndez, Alicia ; Michels, Paul ; Miotto, Giovanni ; Mistiaen, Wilhelm P. ; Mizushima, Noboru ; Mograbi, Baharia ; Monastyrska, Iryna ; Moore, Michael N. ; Moreira, Paula I. ; Moriyasu, Yuji ; Motyl, Tomasz ; Münz, Christian ; Murphy, Leon O. ; Naqvi, Naweed I. ; Neufeld, Thomas P. ; Nishino, Ichizo ; Nixon, Ralph A. ; Noda, Takeshi ; Nürnberg, 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 ; Proikascezanne, Tassula ; Raben, Nina ; Rami, Abdelhaq ; Reggiori, Fulvio ; Rohrer, Bärbel ; 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 C M ; Simon, Hans Uwe ; Simone, Crisfiano ; 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. ; Tallóczy, 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 ; Uzcátegui, Néstor L. ; Van Der Klei, Ida ; Vaquero, Eva C. ; Vellai, Tibor ; Vogel, Michael W. ; Wang, Hong Gang ; Webster, Paul ; Wiley, John W. ; Xi, Zhijun ; Xiao, Gutian ; Yahalom, Joachim ; Yang, Jin Ming ; Yap, George ; Yin, Xiao-Ming ; Yoshimori, Tamotsu ; Yu, Li ; 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, No. 2. pp. 151-175.
@article{751e450d0d524677a4567e08a8fc511a,
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.",
keywords = "Autolysosome, Autophagosome, Flux, Lysosome, Phagophore, Stress, Vacuole",
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 Janice Blum 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 Chin, {Lih Shen} and Augustine Choi and Chu, {Charleen T.} and Jongkyeong Chung and Clarke, {Peter G H} and Clark, {Robert S B} and Clarke, {Steven G.} and Corinne Clav{\'e} and Cleveland, {John L.} and Patrice Codogno and Colombo, {Mar{\'i}a I.} and Ana Cotomontes and Cregg, {James M.} and Cuervo, {Ana Maria} and Jayanta Debnath and Francesca Demarchi and Dennis, {Patrick B.} and Dennis, {Phillip A.} and Vojo Deretic and Devenish, {Rodney J.} and {Di Sano}, Federica and Dice, {J. Fred} and Marian DiFiglia and Savithramma Dinesh-Kumar and Distelhorst, {Clark W.} and Mojgan Djavaheri-Mergny and Dorsey, {Frank C.} and Wulf Dr{\"o}ge and Michel Dron and Dunn, {William A.} and Michael Duszenko and Eissa, {N. Tony} and Zvulun Elazar and Audrey Esclatine and Eskelinen, {Eeva Liisa} and L{\'a}szl{\'o} F{\'e}s{\"u}s and Finley, {Kim D.} and Fuentes, {Jos{\'e} M.} and Juan Fueyo and Kozo Fujisaki and Brigitte Galliot and Gao, {Fen Biao} and Gewirtz, {David A.} and Gibson, {Spencer B.} and Antje Gohla and Goldberg, {Alfred L.} and Ramon Gonzalez and Cristina Gonz{\'a}lez-Est{\'e}vez and Sharon Gorski and Gottlieb, {Roberta A.} and Dieter H{\"a}ussinger and He, {You Wen} and Kim Heidenreich and Hill, {Joseph A.} and Maria H{\o}yer-Hansen and Xun Hu and Huang, {Wei Pang} and Akiko Iwasaki and Marja J{\"a}{\"a}ttel{\"a} 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 A K W} and Hong, {Pyo Kim} and Adi Kimchi and Kinsella, {Timothy J.} and Kirill Kiselyov and Katsuhiko Kitamoto and Erwin Knecht and Masaaki Komatsu and Eiki Kominami and Seiji Kondo and Kov{\'a}cs, {Attila L.} and Guido Kroemer and Kuan, {Chia Yi} and Rakesh Kumar and Mondira Kundu and Jacques Landry and Marianne Laporte and Weidong Le and Lei, {Huan Yao} and Lenardo, {Michael J.} and Beth Levine and Andrew Lieberman and Lim, {Kah Leong} and Lin, {Fu Cheng} and Willisa Liou and Liu, {Leroy F.} and Gabriel Lopez-Berestein and Carlos L{\'o}pez-Ot{\'i}n 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 Mel{\'e}ndez and Paul Michels and Giovanni Miotto and Mistiaen, {Wilhelm P.} and Noboru Mizushima and Baharia Mograbi and Iryna Monastyrska and Moore, {Michael N.} and Moreira, {Paula I.} and Yuji Moriyasu and Tomasz Motyl and Christian M{\"u}nz and Murphy, {Leon O.} and Naqvi, {Naweed I.} and Neufeld, {Thomas P.} and Ichizo Nishino and Nixon, {Ralph A.} and Takeshi Noda and Bernd N{\"u}rnberg 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 Proikascezanne and Nina Raben and Abdelhaq Rami and Fulvio Reggiori and B{\"a}rbel 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 C M} and Simon, {Hans Uwe} and Crisfiano 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 Tall{\'o}czy 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 Uzc{\'a}tegui, {N{\'e}stor L.} and {Van Der Klei}, Ida and Vaquero, {Eva C.} and Tibor Vellai and Vogel, {Michael W.} and Wang, {Hong Gang} and Paul Webster and Wiley, {John W.} and Zhijun Xi and Gutian Xiao and Joachim Yahalom and Yang, {Jin Ming} and George Yap and Xiao-Ming Yin and Tamotsu Yoshimori and Li Yu and Zhenyu Yue and Michisuke Yuzaki and Olga Zabirnyk and Xiaoxiang Zheng and Xiongwei Zhu and Deter, {Russell L.}",
year = "2008",
month = "2",
day = "16",
language = "English",
volume = "4",
pages = "151--175",
journal = "Autophagy",
issn = "1554-8627",
publisher = "Landes Bioscience",
number = "2",

}

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

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 - Clarke, Peter G H

AU - Clark, Robert S B

AU - Clarke, Steven G.

AU - Clavé, Corinne

AU - Cleveland, John L.

AU - Codogno, Patrice

AU - Colombo, María I.

AU - Cotomontes, Ana

AU - Cregg, James M.

AU - Cuervo, Ana Maria

AU - Debnath, Jayanta

AU - Demarchi, Francesca

AU - Dennis, Patrick B.

AU - Dennis, Phillip A.

AU - Deretic, Vojo

AU - Devenish, Rodney J.

AU - Di Sano, Federica

AU - Dice, J. Fred

AU - DiFiglia, Marian

AU - Dinesh-Kumar, Savithramma

AU - Distelhorst, Clark W.

AU - Djavaheri-Mergny, Mojgan

AU - Dorsey, Frank C.

AU - Dröge, Wulf

AU - Dron, Michel

AU - Dunn, William A.

AU - Duszenko, Michael

AU - Eissa, N. Tony

AU - Elazar, Zvulun

AU - Esclatine, Audrey

AU - Eskelinen, Eeva Liisa

AU - Fésüs, László

AU - Finley, Kim D.

AU - Fuentes, José 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 - González-Estévez, Cristina

AU - Gorski, Sharon

AU - Gottlieb, Roberta A.

AU - Häussinger, Dieter

AU - He, You Wen

AU - Heidenreich, Kim

AU - Hill, Joseph A.

AU - Høyer-Hansen, Maria

AU - Hu, Xun

AU - Huang, Wei Pang

AU - Iwasaki, Akiko

AU - Jäättelä, 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 A K W

AU - Hong, Pyo Kim

AU - Kimchi, Adi

AU - Kinsella, Timothy J.

AU - Kiselyov, Kirill

AU - Kitamoto, Katsuhiko

AU - Knecht, Erwin

AU - Komatsu, Masaaki

AU - Kominami, Eiki

AU - Kondo, Seiji

AU - Kovács, 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 - Lenardo, Michael J.

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 - López-Otín, 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 - Meléndez, Alicia

AU - Michels, Paul

AU - Miotto, Giovanni

AU - Mistiaen, Wilhelm P.

AU - Mizushima, Noboru

AU - Mograbi, Baharia

AU - Monastyrska, Iryna

AU - Moore, Michael N.

AU - Moreira, Paula I.

AU - Moriyasu, Yuji

AU - Motyl, Tomasz

AU - Münz, Christian

AU - Murphy, Leon O.

AU - Naqvi, Naweed I.

AU - Neufeld, Thomas P.

AU - Nishino, Ichizo

AU - Nixon, Ralph A.

AU - Noda, Takeshi

AU - Nürnberg, 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 - Proikascezanne, Tassula

AU - Raben, Nina

AU - Rami, Abdelhaq

AU - Reggiori, Fulvio

AU - Rohrer, Bärbel

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 C M

AU - Simon, Hans Uwe

AU - Simone, Crisfiano

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 - Tallóczy, 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 - Uzcátegui, Néstor 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 - Wiley, John W.

AU - Xi, Zhijun

AU - Xiao, Gutian

AU - Yahalom, Joachim

AU - Yang, Jin Ming

AU - Yap, George

AU - Yin, Xiao-Ming

AU - Yoshimori, Tamotsu

AU - Yu, Li

AU - Yue, Zhenyu

AU - Yuzaki, Michisuke

AU - Zabirnyk, Olga

AU - Zheng, Xiaoxiang

AU - Zhu, Xiongwei

AU - Deter, Russell L.

PY - 2008/2/16

Y1 - 2008/2/16

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.

KW - Autolysosome

KW - Autophagosome

KW - Flux

KW - Lysosome

KW - Phagophore

KW - Stress

KW - Vacuole

UR - http://www.scopus.com/inward/record.url?scp=38949108670&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=38949108670&partnerID=8YFLogxK

M3 - Article

VL - 4

SP - 151

EP - 175

JO - Autophagy

JF - Autophagy

SN - 1554-8627

IS - 2

ER -