Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Autophagy, Journal Year: 2016, Volume and Issue: 12(1), P. 1 - 222

Published: Jan. 2, 2016

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, on this topic has continued to accelerate, and many new scientists have entered field. Our knowledge base relevant technologies also been expanding. Accordingly, it is important update these monitoring autophagy different organisms. Various reviews described range assays that used purpose. Nevertheless, there continues be confusion regarding acceptable methods measure autophagy, especially multicellular eukaryotes. For example, a key point needs emphasized difference between measurements monitor numbers or volume autophagic elements (e.g., autophagosomes autolysosomes) at any stage process versus those flux through pathway (i.e., complete including amount rate cargo sequestered degraded). particular, block macroautophagy results autophagosome accumulation must differentiated from stimuli increase activity, defined as increased induction coupled with delivery to, degradation within, lysosomes (in most higher eukaryotes some protists such Dictyostelium) vacuole plants fungi). other words, investigators field understand appearance more does not necessarily equate fact, cases, accumulate because trafficking without concomitant change biogenesis, whereas an autolysosomes may reflect reduction degradative activity. It worth emphasizing here lysosomal digestion evaluating its competence crucial part evaluation flux, autophagy. Here, present selection interpretation use by who aim examine related processes, well reviewers need provide realistic reasonable critiques papers are focused processes. These meant formulaic rules, appropriate depend question being asked system used. addition, emphasize no individual assay guaranteed one every situation, strongly recommend multiple Along lines, potential pleiotropic effects due blocking genetic manipulation, imperative target gene knockout RNA interference than autophagy-related protein. Atg proteins, groups involved cellular pathways implying all proteins can specific marker process. guidelines, consider various assessing what information can, cannot, obtained them. Finally, discussing merits limits particular assays, hope encourage technical innovation

Language: Английский

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AMPK: a nutrient and energy sensor that maintains energy homeostasis

Nature Reviews Molecular Cell Biology, Journal Year: 2012, Volume and Issue: 13(4), P. 251 - 262

Published: March 22, 2012

Language: Английский

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Guidelines for the use and interpretation of assays for monitoring autophagy

Autophagy, Journal Year: 2012, Volume and Issue: 8(4), P. 445 - 544

Published: April 1, 2012

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, on this topic has continued to accelerate, and many new scientists have entered field. Our knowledge base relevant technologies also been expanding. Accordingly, it is important update these monitoring autophagy different organisms. Various reviews described range assays that used purpose. Nevertheless, there continues be confusion regarding acceptable methods measure autophagy, especially multicellular eukaryotes. A key point needs emphasized a difference between measurements monitor numbers or volume autophagic elements (e.g., autophagosomes autolysosomes) at any stage process vs. those flux through pathway (i.e., complete process); thus, block macroautophagy results autophagosome accumulation differentiated from stimuli result increased activity, defined as induction coupled with delivery to, degradation within, lysosomes (in most higher eukaryotes some protists such Dictyostelium) vacuole plants fungi). other words, investigators field understand appearance more does not necessarily equate fact, cases, accumulate because trafficking without concomitant change biogenesis, whereas an increase autolysosomes may reflect reduction degradative activity. Here, present selection interpretation use by who aim examine related processes, well reviewers need provide realistic reasonable critiques papers are focused processes. These meant formulaic rules, appropriate depend part question being asked system used. addition, emphasize no individual assay guaranteed one every situation, strongly recommend multiple guidelines, consider various assessing what information can, cannot, obtained them. Finally, discussing merits limits particular assays, hope encourage technical innovation

Language: Английский

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Mitochondrial Fission, Fusion, and Stress

Science, Journal Year: 2012, Volume and Issue: 337(6098), P. 1062 - 1065

Published: Aug. 30, 2012

Mitochondrial fission and fusion play critical roles in maintaining functional mitochondria when cells experience metabolic or environmental stresses. Fusion helps mitigate stress by mixing the contents of partially damaged as a form complementation. Fission is needed to create new mitochondria, but it also contributes quality control enabling removal can facilitate apoptosis during high levels cellular stress. Disruptions these processes affect normal development, they have been implicated neurodegenerative diseases, such Parkinson's.

Language: Английский

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AMPK: guardian of metabolism and mitochondrial homeostasis

Nature Reviews Molecular Cell Biology, Journal Year: 2017, Volume and Issue: 19(2), P. 121 - 135

Published: Oct. 4, 2017

Language: Английский

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TFEB Links Autophagy to Lysosomal Biogenesis

Science, Journal Year: 2011, Volume and Issue: 332(6036), P. 1429 - 1433

Published: May 27, 2011

Starvation activates a transcriptional program controlling autophagosome formation, lysosome fusion, and substrate degradation.

Language: Английский

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The Role of Atg Proteins in Autophagosome Formation

Annual Review of Cell and Developmental Biology, Journal Year: 2011, Volume and Issue: 27(1), P. 107 - 132

Published: March 3, 2011

Macroautophagy is mediated by a unique organelle, the autophagosome, which encloses portion of cytoplasm for delivery to lysosome. Autophagosome formation dynamically regulated starvation and other stresses involves complicated membrane reorganization. Since discovery yeast Atg-related proteins, autophagosome has been dissected at molecular level. In this review we describe mechanism with particular focus on function Atg proteins long-standing discussion regarding origin membrane.

Language: Английский

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The AMPK signalling pathway coordinates cell growth, autophagy and metabolism

Nature Cell Biology, Journal Year: 2011, Volume and Issue: 13(9), P. 1016 - 1023

Published: Sept. 1, 2011

Language: Английский

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Biological Functions of Autophagy Genes: A Disease Perspective

Cell, Journal Year: 2019, Volume and Issue: 176(1-2), P. 11 - 42

Published: Jan. 1, 2019

Language: Английский

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An Overview of Autophagy: Morphology, Mechanism, and Regulation

Antioxidants and Redox Signaling, Journal Year: 2013, Volume and Issue: 20(3), P. 460 - 473

Published: June 3, 2013

Significance: Autophagy is a highly conserved eukaryotic cellular recycling process. Through the degradation of cytoplasmic organelles, proteins, and macromolecules, breakdown products, autophagy plays important roles in cell survival maintenance. Accordingly, dysfunction this process contributes to pathologies many human diseases. Recent Advances: Extensive research currently being done better understand autophagy. In review, we describe current knowledge morphology, molecular mechanism, regulation mammalian Critical Issues: At mechanistic regulatory levels, there are still unanswered questions points confusion that have yet be resolved. Future Directions: further research, more complete accurate picture mechanism will not only strengthen our understanding significant process, but aid development new treatments for diseases which functioning properly. Antioxid. Redox Signal. 20, 460–473.

Language: Английский

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