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

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 - 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 - Coto-Montes, 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 - Kim, Hong Pyo

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 - Proikas-Cezanne, 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, Miklós

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, 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 - 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

Y1 - 2008/2

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 - Animals

KW - Autophagy/physiology

KW - Autophagy-Related Protein 8 Family

KW - Clinical Laboratory Techniques

KW - Data Interpretation, Statistical

KW - Eukaryotic Cells/physiology

KW - Guidelines as Topic

KW - Humans

KW - Microscopy, Fluorescence/methods

KW - Microtubule-Associated Proteins/metabolism

KW - Models, Biological

KW - Phagosomes/metabolism

KW - Plants/metabolism

KW - Protein Processing, Post-Translational

KW - Protein Transport

KW - Saccharomyces cerevisiae Proteins/metabolism

U2 - 10.4161/auto.5338

DO - 10.4161/auto.5338

M3 - Review

C2 - 18188003

SN - 1554-8627

VL - 4

SP - 151

EP - 175

JO - Autophagy

JF - Autophagy

IS - 2

ER -