Guidelines for the use and interpretation of assays for monitoring autophagy

doi:10.4161/auto.19496

CORC > 武汉植物园 > 中国科学院武汉植物园

	Guidelines for the use and interpretation of assays for monitoring autophagy
	Klionsky, Daniel J.858; Abdalla, Fabio C.861; Abeliovich, Hagai 862; Abraham, Robert T.863; Acevedo-Arozena, Abraham 864; Adeli, Khosrow 865; Agholme, Lotta 866; Agnello, Maria 867; Agostinis, Patrizia 868; Aguirre-Ghiso, Julio A.869,870
刊名	AUTOPHAGY
	2012-04-01
卷号	8 期号:4 页码:445-544
关键词	autolysosome autophagosome flux LC3 lysosome phagophore stress vacuole
ISSN号	1554-8627
DOI	10.4161/auto.19496
英文摘要	In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are 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 monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
资助项目	National Institutes of Health Public Health Service[GM53396]
WOS研究方向	Cell Biology
语种	英语
出版者	LANDES BIOSCIENCE
WOS记录号	WOS:000305403400002
内容类型	期刊论文
源URL	[http://202.127.146.157/handle/2RYDP1HH/3866]
专题	中国科学院武汉植物园
通讯作者	Klionsky, Daniel J.
作者单位	1.Dulbecco Telethon Inst, Padua, Italy 2.Univ Milan, Dept Biomol Sci & Biotechnol, Milan, Italy 3.Buck Inst Res Aging, Novato, CA USA 4.Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Stockholm, Sweden 5.Harvard Univ, Sch Med, Dept Genet, Boston, MA USA 6.Univ Texas MD Anderson Canc Ctr, Dept Melanoma Med Oncol, Houston, TX 77030 USA 7.Penn State Coll Med, Penn State Hershey Med Ctr, Dept Med Hematol Oncol, Lab Translat Oncol & Expt Canc Therapeut, Hershey, PA USA 8.Weizmann Inst Sci, Dept Biol Chem, IL-76100 Rehovot, Israel 9.Univ Milan, European Inst Oncol, Milan, Italy 10.Univ Stellenbosch, Dept Physiol Sci, ZA-7600 Stellenbosch, South Africa
推荐引用方式 GB/T 7714	Klionsky, Daniel J.,Abdalla, Fabio C.,Abeliovich, Hagai,et al. Guidelines for the use and interpretation of assays for monitoring autophagy[J]. AUTOPHAGY,2012,8(4):445-544.
APA	Klionsky, Daniel J..,Abdalla, Fabio C..,Abeliovich, Hagai.,Abraham, Robert T..,Acevedo-Arozena, Abraham.,...&Sheng, Zu-Hang.(2012).Guidelines for the use and interpretation of assays for monitoring autophagy.AUTOPHAGY,8(4),445-544.
MLA	Klionsky, Daniel J.,et al."Guidelines for the use and interpretation of assays for monitoring autophagy".AUTOPHAGY 8.4(2012):445-544.