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Article: Guidelines for the use and interpretation of assays for monitoring autophagy

TitleGuidelines for the use and interpretation of assays for monitoring autophagy
Authors
Klionsky, DJAbdalla, FCAbeliovich, HAbraham, RTAcevedoArozena, AAdeli, KAgholme, LAgnello, MAgostinis, PAguirreGhiso, JAAhn, HJLe Cam, LApostolova, NFurukawa, RHGaestel, MGailly, PGajewska, MNogalska, AGalliot, BGaly, VGanesh, SSpringer, WBeau, IGanetzky, BGanley, IGGao, FBArancia, GGao, GFGao, JTettamanti, GGarcia, LGarciaManero, GGarciaMarcos, MTaegtmeyer, HGarmyn, MBultynck, GGartel, ALGatti, EGautel, MGawriluk, TRAris, JPBortoluci, KRGegg, MEGeng, JLedbetter, DJGermain, MGestwicki, JEGewirtz, DASrinivasula, SMGhavami, SGhosh, PGiammarioli, AMGiatromanolaki, ANNoguchi, SGibson, SBLee, AJXAsanuma, KGilkerson, RWGinger, MLGinsberg, HNGolab, JStefanis, LGoligorsky, MSGolstein, PGomezManzano, CNotterpek, LLee, BWGoncu, EGongora, CGonzalez, CDAsare, NYOGonzalez, RGonzálezEstévez, CGonzálezPolo, RASteffan, JSGonzalezRey, EGorbunov, NVLee, GMNovak, IGorski, SGoruppi, SGottlieb, RAGozuacik, DGranato, GEAshida, HGrant, GDGreen, KNStendel, RLee, JGregorc, ANozaki, TGros, FGrose, CGrunt, TWGual, PGuan, JLGuan, KLGuichard, SMAskanas, VLee, JHGukovskaya, ASStenmark, HNukina, NGukovsky, IGunst, JGustafsson, ABHalayko, AJHale, ANHalonen, SKHamasaki, MLee, MHan, FHan, TAskew, DSNürnberger, TStephanou, AHancock, MKHansen, MHarada, HHarada, MHardt, SEOsborne, TFPagnini, UHarper, JWHarris, ALHarris, JHarris, SDNyfeler, BHashimoto, MStern, STAuberger, PHaspel, JAHayashi, SILee, MSHazelhurst, LAHe, CHe, YWHébert, MJHeidenreich, KAObara, KHelfrich, MHHelgason, GVSternberg, CHenske, EPLee, SHBaba, MHerman, BHerman, PKHetz, CHilfiker, SHill, JAOberley, TDHocking, LJHofman, PHofmann, TGLeeuwenburgh, CStork, BHöhfeld, JHolyoake, TLBackues, SKHong, MHHood, DAHotamisligil, GSThevissen, KHouwerzijl, EJHøyerHansen, MTafani, MHu, BHu, CAAStrålfors, PHu, HMHua, YHuang, CBaehrecke, EHHuang, JOddo, SHuang, SBéchet, DMHuang, WPHuber, TBHuh, WKHung, THBurbulla, LFHupp, TRHur, GMHurley, JBHussain, SNABossis, ILegembre, PBahr, BAHussey, PJHwang, JJHwang, SIchihara, AIlkhanizadeh, SSubauste, CSInoki, KInto, TIovane, VLegouis, ROgawa, MIovanna, JLIp, NYBai, XYIsaka, YIshida, HIsidoro, CIsobe, KISui, XIwasaki, ALehmann, MIzquierdo, MOhashi, TIzumi, YJaakkola, PMJäättelä, MJackson, GRBailly, YJackson, WTJanji, BJendrach, MLei, HYSulzer, DJeon, JHOkamoto, KJeung, EBJiang, HJiang, HJiang, JXJiang, MJiang, QBaiocchi, RKim, HRLei, QYJiang, XJiménez, ASun, JOleinick, NLJin, MJin, SJoe, COJohansen, TJohnson, DEJohnson, GVWTavernarakis, NJones, NLJoseph, BBaldini, GJoseph, SKOliver, FJSun, SYJoubert, AMJuhász, GJuilleratJeanneret, LJung, CHThorburn, AJung, YKKaarniranta, KKaasik, AKabuta, TKadowaki, MOlsen, LJBalduini, WSun, ZJKagedal, KKamada, YLeib, DAKaminskyy, VOKampinga, HHKanamori, HKang, CKang, KBIl Kang, KOlsson, SKang, RKang, YASung, JJYLeiro, JBallabio, AKanki, TKanneganti, TDKanno, HKanthasamy, AGKanthasamy, AKarantza, VOpota, OKaushal, GPKaushik, SLemasters, JJKawazoe, YSuzuki, KKe, PYBamber, BAKehrl, JHKelekar, AKerkhoff, CKessel, DHThompson, CBKhalil, HLemoine, AKiel, 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FEDe Felici, MDe Groot, JFDe Haan, CAMDe Martino, LLandry, JDe Milito, ADe Tata, VNetea, MGDebnath, JSlack, RSDegterev, ADehay, BAlmonteBecerril, MDelbridge, LMDDemarchi, FBay, BHDeng, YZDengjel, JDent, PNeufeld, TPDenton, DDeretic, VSmaili, SSDesai, SDDevenish, RJDi Gioacchino, MOzpolat, BAmano, ADi Paolo, GDi Pietro, CDíazAraya, GBornhauser, BCDíazLaviada, IDiazMeco, MTDiazNido, JBultman, SJDikic, ILane, JDDineshKumar, SPDing, WXDistelhorst, CWAmaravadi, RDiwan, ATerman, ADjavaheriMergny, MDokudovskaya, SDong, ZDorsey, FCTabas, ISmith, DRDosenko, VDowling, JJDoxsey, SDreux, MDrew, MENey, PAAmarnath, SDuan, QDuchosal, MALapaquette, PDuff, KDugail, ISoengas, MSDurbeej, MDuszenko, MEdelstein, CLEdinger, ALNezis, IPEgea, GEichinger, LLaporte, JFAmer, AOEissa, NTEkmekcioglu, SElDeiry, WSSoldati, TElazar, ZElgendy, MEllerby, LMNguyen, HPEr Eng, KLászló, LEngelbrecht, AMEngelender, SErenpreisa, JAndrieuAbadie, NEscalante, REsclatine, ASong, XEskelinen, ELEspert, LNie, DLavandero, SEspina, VFan, HFan, JFan, QWFan, ZFang, SAnantharam, VFang, YSood, AKFanto, MLavoie, JNNishino, IFanzani, AFarkas, TFarré, JCFaure, MFechheimer, MFeng, CGFeng, JFeng, QAnn, DKLayfield, RSoong, TWNislow, CFeng, YFésüs, LFeuer, RFigueiredoPereira, MEFimia, GMFingar, DCFinkbeiner, SFinkel, TLazo, PAFinley, KDFiorito, FNixon, RASotgia, FAnoopkumarDukie, SFisher, EAFisher, PBFlajolet, MFlorezMcClure, MLFlorio, SLe, WFon, EAFornai, FFortunato, FNoda, TFotedar, RSpector, SAFowler, DHAoki, HFox, HSFranco, RPaganetti, PFrankel, LBFransen, MFuentes, JMFueyo, JNoegel, AAFujii, JFujisaki, KSpies, CDFujita, EFukuda, M
KeywordsAutolysosome
Autophagosome
Flux
LC3
Lysosome
Phagophore
Stress
Vacuole
Issue Date2012
Citation
Autophagy, 2012, v. 8 n. 4, p. 445-544 How to Cite?
AbstractIn 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. © 2012 Landes Bioscience.
Persistent Identifierhttp://hdl.handle.net/10722/168634
ISSN
2023 Impact Factor: 14.6
2023 SCImago Journal Rankings: 4.035
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorKlionsky, DJen_HK
dc.contributor.authorAbdalla, FCen_HK
dc.contributor.authorAbeliovich, Hen_HK
dc.contributor.authorAbraham, RTen_HK
dc.contributor.authorAcevedoArozena, Aen_HK
dc.contributor.authorAdeli, Ken_HK
dc.contributor.authorAgholme, Len_HK
dc.contributor.authorAgnello, Men_HK
dc.contributor.authorAgostinis, Pen_HK
dc.contributor.authorAguirreGhiso, JAen_HK
dc.contributor.authorAhn, HJen_HK
dc.contributor.authorLe Cam, Len_HK
dc.contributor.authorApostolova, Nen_HK
dc.contributor.authorFurukawa, RHen_HK
dc.contributor.authorGaestel, Men_HK
dc.contributor.authorGailly, Pen_HK
dc.contributor.authorGajewska, Men_HK
dc.contributor.authorNogalska, Aen_HK
dc.contributor.authorGalliot, Ben_HK
dc.contributor.authorGaly, Ven_HK
dc.contributor.authorGanesh, Sen_HK
dc.contributor.authorSpringer, Wen_HK
dc.contributor.authorBeau, Ien_HK
dc.contributor.authorGanetzky, Ben_HK
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dc.contributor.authorGreen, KNen_HK
dc.contributor.authorStendel, Ren_HK
dc.contributor.authorLee, Jen_HK
dc.contributor.authorGregorc, Aen_HK
dc.contributor.authorNozaki, Ten_HK
dc.contributor.authorGros, Fen_HK
dc.contributor.authorGrose, Cen_HK
dc.contributor.authorGrunt, TWen_HK
dc.contributor.authorGual, Pen_HK
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dc.contributor.authorNukina, Nen_HK
dc.contributor.authorGukovsky, Ien_HK
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dc.contributor.authorDugail, Ien_HK
dc.contributor.authorSoengas, MSen_HK
dc.contributor.authorDurbeej, Men_HK
dc.contributor.authorDuszenko, Men_HK
dc.contributor.authorEdelstein, CLen_HK
dc.contributor.authorEdinger, ALen_HK
dc.contributor.authorNezis, IPen_HK
dc.contributor.authorEgea, Gen_HK
dc.contributor.authorEichinger, Len_HK
dc.contributor.authorLaporte, JFen_HK
dc.contributor.authorAmer, AOen_HK
dc.contributor.authorEissa, NTen_HK
dc.contributor.authorEkmekcioglu, Sen_HK
dc.contributor.authorElDeiry, WSen_HK
dc.contributor.authorSoldati, Ten_HK
dc.contributor.authorElazar, Zen_HK
dc.contributor.authorElgendy, Men_HK
dc.contributor.authorEllerby, LMen_HK
dc.contributor.authorNguyen, HPen_HK
dc.contributor.authorEr Eng, Ken_HK
dc.contributor.authorLászló, Len_HK
dc.contributor.authorEngelbrecht, AMen_HK
dc.contributor.authorEngelender, Sen_HK
dc.contributor.authorErenpreisa, Jen_HK
dc.contributor.authorAndrieuAbadie, Nen_HK
dc.contributor.authorEscalante, Ren_HK
dc.contributor.authorEsclatine, Aen_HK
dc.contributor.authorSong, Xen_HK
dc.contributor.authorEskelinen, ELen_HK
dc.contributor.authorEspert, Len_HK
dc.contributor.authorNie, Den_HK
dc.contributor.authorLavandero, Sen_HK
dc.contributor.authorEspina, Ven_HK
dc.contributor.authorFan, Hen_HK
dc.contributor.authorFan, Jen_HK
dc.contributor.authorFan, QWen_HK
dc.contributor.authorFan, Zen_HK
dc.contributor.authorFang, Sen_HK
dc.contributor.authorAnantharam, Ven_HK
dc.contributor.authorFang, Yen_HK
dc.contributor.authorSood, AKen_HK
dc.contributor.authorFanto, Men_HK
dc.contributor.authorLavoie, JNen_HK
dc.contributor.authorNishino, Ien_HK
dc.contributor.authorFanzani, Aen_HK
dc.contributor.authorFarkas, Ten_HK
dc.contributor.authorFarré, JCen_HK
dc.contributor.authorFaure, Men_HK
dc.contributor.authorFechheimer, Men_HK
dc.contributor.authorFeng, CGen_HK
dc.contributor.authorFeng, Jen_HK
dc.contributor.authorFeng, Qen_HK
dc.contributor.authorAnn, DKen_HK
dc.contributor.authorLayfield, Ren_HK
dc.contributor.authorSoong, TWen_HK
dc.contributor.authorNislow, Cen_HK
dc.contributor.authorFeng, Yen_HK
dc.contributor.authorFésüs, Len_HK
dc.contributor.authorFeuer, Ren_HK
dc.contributor.authorFigueiredoPereira, MEen_HK
dc.contributor.authorFimia, GMen_HK
dc.contributor.authorFingar, DCen_HK
dc.contributor.authorFinkbeiner, Sen_HK
dc.contributor.authorFinkel, Ten_HK
dc.contributor.authorLazo, PAen_HK
dc.contributor.authorFinley, KDen_HK
dc.contributor.authorFiorito, Fen_HK
dc.contributor.authorNixon, RAen_HK
dc.contributor.authorSotgia, Fen_HK
dc.contributor.authorAnoopkumarDukie, Sen_HK
dc.contributor.authorFisher, EAen_HK
dc.contributor.authorFisher, PBen_HK
dc.contributor.authorFlajolet, Men_HK
dc.contributor.authorFlorezMcClure, MLen_HK
dc.contributor.authorFlorio, Sen_HK
dc.contributor.authorLe, Wen_HK
dc.contributor.authorFon, EAen_HK
dc.contributor.authorFornai, Fen_HK
dc.contributor.authorFortunato, Fen_HK
dc.contributor.authorNoda, Ten_HK
dc.contributor.authorFotedar, Ren_HK
dc.contributor.authorSpector, SAen_HK
dc.contributor.authorFowler, DHen_HK
dc.contributor.authorAoki, Hen_HK
dc.contributor.authorFox, HSen_HK
dc.contributor.authorFranco, Ren_HK
dc.contributor.authorPaganetti, Pen_HK
dc.contributor.authorFrankel, LBen_HK
dc.contributor.authorFransen, Men_HK
dc.contributor.authorFuentes, JMen_HK
dc.contributor.authorFueyo, Jen_HK
dc.contributor.authorNoegel, AAen_HK
dc.contributor.authorFujii, Jen_HK
dc.contributor.authorFujisaki, Ken_HK
dc.contributor.authorSpies, CDen_HK
dc.contributor.authorFujita, Een_HK
dc.contributor.authorFukuda, Men_HK
dc.date.accessioned2012-10-08T03:23:42Z-
dc.date.available2012-10-08T03:23:42Z-
dc.date.issued2012en_HK
dc.identifier.citationAutophagy, 2012, v. 8 n. 4, p. 445-544en_HK
dc.identifier.issn1554-8627en_HK
dc.identifier.urihttp://hdl.handle.net/10722/168634-
dc.description.abstractIn 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. © 2012 Landes Bioscience.en_HK
dc.languageengen_US
dc.relation.ispartofAutophagyen_HK
dc.subjectAutolysosomeen_HK
dc.subjectAutophagosomeen_HK
dc.subjectFluxen_HK
dc.subjectLC3en_HK
dc.subjectLysosomeen_HK
dc.subjectPhagophoreen_HK
dc.subjectStressen_HK
dc.subjectVacuoleen_HK
dc.titleGuidelines for the use and interpretation of assays for monitoring autophagyen_HK
dc.typeArticleen_HK
dc.identifier.emailChe, CM: cmche@hku.hken_HK
dc.identifier.emailChye, ML: mlchye@hkucc.hku.hken_HK
dc.identifier.emailLok, CN: cnlok@hku.hken_HK
dc.identifier.emailSy, LK: sylk@hku.hken_HK
dc.identifier.authorityChe, CM=rp00670en_HK
dc.identifier.authorityChye, ML=rp00687en_HK
dc.identifier.authorityLok, CN=rp00752en_HK
dc.identifier.authoritySy, LK=rp00784en_HK
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.doi10.4161/auto.19496en_HK
dc.identifier.pmid22966490-
dc.identifier.scopuseid_2-s2.0-84862295360en_HK
dc.identifier.hkuros203531-
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dc.identifier.volume8en_HK
dc.identifier.issue4en_HK
dc.identifier.spage445en_HK
dc.identifier.epage544en_HK
dc.identifier.isiWOS:000305403400002-
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dc.identifier.issnl1554-8627-

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