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

@article{Klionsky2008GuidelinesFT,
  title={Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes},
  author={Daniel J. Klionsky and Hagai Abeliovich and Patrizia Agostinis and Devendra K. Agrawal and Gjumrakch Aliev and David S. Askew and Misuzu Baba and Eric H. Baehrecke and Ben A Bahr and Andrea Ballabio and Bruce A Bamber and Diane C. Bassham and Ettore Bergamini and Xiaoning Bi and Martine Biard-Piechaczyk and Janice S Blum and Dale E. Bredesen and Jeffrey L. Brodsky and John H. Brumell and Ulf T. Brunk and Wilfried Bursch and Nadine Camougrand and Eduardo Cebollero and Francesco Cecconi and Yingyu Chen and Lih-Shen Chin and Augustine M K Choi and Charleen T. Chu and Jongkyeong Chung and Peter G. H. Clarke and Robert S. B. Clark and Steven G. Clarke and Corinne Clav{\'e} and John L. Cleveland and Patrice Codogno and Mar{\'i}a Isabel Colombo and Ana Coto-Montes and James Michael Cregg and Ana Mar{\'i}a Cuervo and Jayanta Debnath and Francesca Demarchi and Patrick B. Dennis and Phillip A. Dennis and Vojo Deretic and Rodney J Devenish and Federica Di Sano and James Fred Dice and Marian DiFiglia and Savithramma P. Dinesh-Kumar and Clark W. Distelhorst and Mojgan Djavaheri-Mergny and Frank C. Dorsey and Wulf Dröge and Michel Dron and W Augustine Dunn and Michael Duszenko and N Tony Eissa and Zvulun Elazar and Audrey Esclatine and Eeva-Liisa Eskelinen and L{\'a}szl{\'o} F{\'e}s{\"u}s and Kim D. Finley and Jos{\'e} M. Fuentes and Juan Fueyo and Kozo Fujisaki and Brigitte Galliot and Fen-Biao Gao and David A. Gewirtz and Spencer B. Gibson and Antje Gohla and Alfred L. Goldberg and Ram{\'o}n Gonzalez and Cristina Gonz{\'a}lez-Est{\'e}vez and Sharon M. Gorski and Roberta A. Gottlieb and Dieter H{\"a}ussinger and You-Wen He and Kim A. Heidenreich and Joseph A. Hill and Maria H{\o}yer-Hansen and Xun Hu and Wei‐Pang Huang and Akiko Iwasaki and Marja J{\"a}{\"a}ttel{\"a} and William T. Jackson and Xuejun Jiang and Shengkan Jin and Terje Johansen and Jae U. Jung and Motoni Kadowaki and Chanhee Kang and Ameeta Kelekar and David Kessel and Jan A. K. W. Kiel and Hong-pyo Kim and Adi Kimchi and Timothy J. Kinsella and Kirill Kiselyov and Katsuhiko Kitamoto and Erwin Knecht and Masaaki Komatsu and Eiki Kominami and Seiji Kondo and Attila Lajos Kov{\'a}cs and Guido Kroemer and Chia-Yi Kuan and Rakesh Kumar and Mondira Kundu and Jacques Landry and Marianne Laporte and Weidong Le and Huan-Yao Lei and Michael J. Lenardo and Beth Levine and Andrew P. Lieberman and Kah-Leong Lim and Fu-cheng Lin and Willisa Liou and Leroy F. Liu and Gabriel Lopez-Berestein and Carlos L{\'o}pez-Ot{\'i}n and Bo Lu and Kay F Macleod and Walter Malorni and Wim Martinet and Ken Matsuoka and Josef Mautner and Alfred J. Meijer and Alicia Meléndez and Paul Michels and Giovanni Miotto and Wilhelm P Mistiaen and Noboru Mizushima and Baharia Mograbi and Iryna Monastyrska and Michael N Moore and Paula I. Moreira and Yuji Moriyasu and Tomasz Motyl and Christian M{\"u}nz and Leon O. Murphy and Naweed I. Naqvi and Thomas P. Neufeld and Ichizo Nishino and Ralph A. Nixon and Takeshi Noda and Bernd N{\"u}rnberg and Michinaga Ogawa and Nancy Oleinick and Laura j. Olsen and Bulent Ozpolat and Shoshana Paglin and Glen E. Palmer and Issidora S Papassideri and Miles Parkes and David H. Perlmutter and George Perry and Mauro Piacentini and Ronit Pinkas-Kramarski and Mark C. Prescott and Tassula Proikas-Cezanne and Nina Raben and Abdelhaq Rami and Fulvio Reggiori and B{\"a}rbel Rohrer and David C. Rubinsztein and Kevin M. Ryan and Junichi Sadoshima and Hiroshi Sakagami and Yasuyoshi Sakai and Marco Sandri and Chihiro Sasakawa and Mikl{\'o}s Sass and Claudio Schneider and Per O. Seglen and Oleksandr Seleverstov and Jeffrey Settleman and John Shacka and Irving M. Shapiro and Andrei A. Sibirny and Elaine Cristina Mathias Silva-Zacarin and Hans-Uwe Simon and Cristiano Simone and Anne Simonsen and Mark A. Smith and Katharina Spanel‐Borowski and Vickram Srinivas and Meredith A. Steeves and Harald A Stenmark and Per E. Stromhaug and Carlos S. Subauste and Seiichi Sugimoto and David L. Sulzer and Toshihiko Suzuki and Michele S. Swanson and Ira A Tabas and Fumihiko Takeshita and Nicholas J. Talbot and Zsolt Tallóczy and Keiji Tanaka and Kozo Tanaka and Isei Tanida and Graham S. Taylor and J. Paul Taylor and Alexei Terman and Gianluca Tettamanti and Craig B. Thompson and Michael Thumm and Aviva M Tolkovsky and Sharon A. Tooze and Ray Truant and Lesya V. Tumanovska and Yasuo Uchiyama and Takashi Ueno and N{\'e}stor L Uzc{\'a}tegui and Ida J. van der Klei and Eva C. Vaquero and Tibor Vellai and Michael W Vogel and Hong-Gang Wang and Paul Webster and John W. Wiley and Zhijun Xi and Gutian Xiao and Joachim Yahalom and Jingqing Yang and George S. Yap and Xiao-Ming Yin and Tamotsu Yoshimori and Li Yu and Zhenyu Yue and Michisuke Yuzaki and Olga Zabirnyk and Xiaoxiang Zheng and Xiongwei Zhu and R L Deter},
  journal={Autophagy},
  year={2008},
  volume={4},
  pages={151 - 175}
}
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… 

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Current approaches that can be used to follow the overall process of autophagy, as well as individual steps, from yeast to human are discussed.

Autophagy: assays and artifacts

TLDR
A critical overview of current methodologies to measure autophagy in cells and in animals is presented and the realization that it is not sufficient to monitor static levels of Autophagy but to measure ‘autophagic flux’ has driven the development of new or modified approaches to detecting autophile detection.

Methodological considerations for assessing autophagy modulators: A study with calcium phosphate precipitates

TLDR
It is shown that CPP induces autophagosome synthesis at early time-points, such as 4 h and 6 h, but at 24 h after treatment, the complex consequences of CPP suggest that it is best avoided as a transfection reagent in studies aiming to analyse Autophagy itself, or processes that are modulated by autophagy, like apoptosis.

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