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

Publication ,  Journal Article
Klionsky, DJ; Abeliovich, H; Agostinis, P; Agrawal, DK; Aliev, G; Askew, DS; Baba, M; Baehrecke, EH; Bahr, BA; Ballabio, A; Bamber, BA; Bi, X ...
Published in: Autophagy
February 2008

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.

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Published In

Autophagy

DOI

EISSN

1554-8635

Publication Date

February 2008

Volume

4

Issue

2

Start / End Page

151 / 175

Location

United States

Related Subject Headings

  • Saccharomyces cerevisiae Proteins
  • Protein Transport
  • Protein Processing, Post-Translational
  • Plants
  • Phagosomes
  • Models, Biological
  • Microtubule-Associated Proteins
  • Microscopy, Fluorescence
  • Humans
  • Guidelines as Topic
 

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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. https://doi.org/10.4161/auto.5338
Klionsky, Daniel J., Hagai Abeliovich, Patrizia Agostinis, Devendra K. Agrawal, Gjumrakch Aliev, David S. Askew, Misuzu Baba, et al. “Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes.Autophagy 4, no. 2 (February 2008): 151–75. https://doi.org/10.4161/auto.5338.
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;4(2):151–75.
Klionsky, Daniel J., et al. “Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes.Autophagy, vol. 4, no. 2, Feb. 2008, pp. 151–75. Pubmed, doi:10.4161/auto.5338.
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, Clarke PGH, Clark RSB, Clarke SG, Clavé C, Cleveland JL, Codogno P, Colombo MI, Coto-Montes 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 E-L, Fésüs L, Finley KD, Fuentes JM, Fueyo J, Fujisaki K, Galliot B, Gao F-B, Gewirtz DA, Gibson SB, Gohla A, Goldberg AL, Gonzalez R, González-Estévez C, Gorski S, Gottlieb RA, Häussinger D, He Y-W, Heidenreich K, Hill JA, Høyer-Hansen M, Hu X, Huang W-P, Iwasaki A, Jäättelä M, Jackson WT, Jiang X, Jin S, Johansen T, Jung JU, Kadowaki M, Kang C, Kelekar A, Kessel DH, Kiel JAKW, Kim HP, Kimchi A, Kinsella TJ, Kiselyov K, Kitamoto K, Knecht E, Komatsu M, Kominami E, Kondo S, Kovács AL, Kroemer G, Kuan C-Y, Kumar R, Kundu M, Landry J, Laporte M, Le W, Lei H-Y, Lenardo MJ, Levine B, Lieberman A, Lim K-L, Lin F-C, 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, 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, Tallóczy Z, 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 H-G, Webster P, Wiley JW, Xi Z, Xiao G, Yahalom J, Yang J-M, Yap G, Yin X-M, Yoshimori T, Yu L, Yue Z, Yuzaki M, Zabirnyk O, Zheng X, Zhu X, Deter RL. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy. 2008 Feb;4(2):151–175.

Published In

Autophagy

DOI

EISSN

1554-8635

Publication Date

February 2008

Volume

4

Issue

2

Start / End Page

151 / 175

Location

United States

Related Subject Headings

  • Saccharomyces cerevisiae Proteins
  • Protein Transport
  • Protein Processing, Post-Translational
  • Plants
  • Phagosomes
  • Models, Biological
  • Microtubule-Associated Proteins
  • Microscopy, Fluorescence
  • Humans
  • Guidelines as Topic