Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes Journal Article
| Authors: | Klionsky, D. J.; Abeliovich, H.; Agostinis, P.; Agrawal, D. K.; Aliev, G.; Askew, D. S.; Baba, M.; Baehrecke, E. H.; Bahr, B. A.; Ballabio, A.; Bamber, B. A.; Bassham, D. C.; Bergamini, E.; Bi, X.; Biard-Piechaczyk, M.; Blum, J. S.; Bredesen, D. E.; Brodsky, J. L.; Brumell, J. H.; Brunk, U. T.; Bursch, W.; Camougrand, N.; Cebollero, E.; Cecconi, F.; Chen, Y.; Chin, L. S.; Choi, A.; Chu, C. T.; Chung, J.; Clarke, P. G. H.; Clark, R. S. B.; Clarke, S. G.; Clavé, C.; Cleveland, J. L.; Codogno, P.; Colombo, M. I.; Cotomontes, A.; Cregg, J. M.; Cuervo, A. M.; Debnath, J.; Demarchi, F.; Dennis, P. B.; Dennis, P. A.; Deretic, V.; Devenish, R. J.; Di Sano, F.; Dice, J. F.; DiFiglia, M.; Dinesh-Kumar, S.; Distelhorst, C. W.; Djavaheri-Mergny, M.; Dorsey, F. C.; Dröge, W.; Dron, M.; Dunn, W. A. Jr; Duszenko, M.; Eissa, N. T.; Elazar, Z.; Esclatine, A.; Eskelinen, E. L.; Fésüs, L.; Finley, K. D.; Fuentes, J. M.; Fueyo, J.; Fujisaki, K.; Galliot, B.; Gao, F. B.; Gewirtz, D. A.; Gibson, S. B.; Gohla, A.; Goldberg, A. L.; Gonzalez, R.; González-Estévez, C.; Gorski, S.; Gottlieb, R. A.; Häussinger, D.; He, Y. W.; Heidenreich, K.; Hill, J. A.; Høyer-Hansen, M.; Hu, X.; Huang, W. P.; Iwasaki, A.; Jäättelä, M.; Jackson, W. T.; Jiang, X.; Jin, S.; Johansen, T.; Jung, J. U.; Kadowaki, M.; Kang, C.; Kelekar, A.; Kessel, D. H.; Kiel, J. A. K. W.; Hong, P. K.; Kimchi, A.; Kinsella, T. J.; Kiselyov, K.; Kitamoto, K.; Knecht, E.; Yahalom, J. |
| Article 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. ©2008 Landes Bioscience. |
| Keywords: | controlled study; human cell; review; nonhuman; animal cell; electron microscopy; animals; models, biological; steady state; protein degradation; green fluorescent protein; practice guideline; enzyme activity; assay; data interpretation, statistical; transcription regulation; protein processing, post-translational; eukaryota; western blotting; cell membrane; protein transport; stress; fluorescence microscopy; microscopy, fluorescence; yeast; saccharomyces cerevisiae proteins; autophagy; fractionation; phagosome; fungi; eukaryote; lysosome; guidelines as topic; protein p62; phagosomes; laboratory techniques and procedures; conjugation; cell vacuole; microtubule-associated proteins; target of rapamycin kinase; autophagosome; plants; eukaryotic cells; autolysosome; flux; phagophore; vacuole |
| Journal Title: | Autophagy |
| Volume: | 4 |
| Issue: | 2 |
| ISSN: | 1554-8627 |
| Publisher: | Taylor & Francis Group |
| Date Published: | 2008-02-01 |
| Start Page: | 151 |
| End Page: | 175 |
| Language: | English |
| PUBMED: | 18188003 |
| PROVIDER: | scopus |
| PMCID: | PMC2654259 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 572" - "Export Date: 17 November 2011" - "Source: Scopus" |
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