Quantitation of "autophagic flux" in mature skeletal muscle.

Autophagy
Jeong-Sun JuConrad C Weihl

Abstract

Reliable and quantitative assays to measure in vivo autophagy are essential. Currently, there are varied methods for monitoring autophagy; however, it is a challenge to measure "autophagic flux" in an in vivo model system. Conversion and subsequent degradation of the microtubule-associated protein 1 light chain 3 (MAP1-LC3/LC3) to the autophagosome associated LC3-II isoform can be evaluated by immunoblot. However, static levels of endogenous LC3-II protein may render possible misinterpretations since LC3-II levels can increase, decrease or remain unchanged in the setting of autophagic induction. Therefore, it is necessary to measure LC3-II protein levels in the presence and absence of lysomotropic agents that block the degradation of LC3-II, a technique aptly named the "autophagometer." In order to measure autophagic flux in mouse skeletal muscle, we treated animals with the microtubule depolarizing agent colchicine. Two days of 0.4 mg/kg/day intraperitoneal colchicine blocked autophagosome maturation to autolysosomes and increased LC3-II protein levels in mouse skeletal muscle by >100%. The addition of an autophagic stimulus such as dietary restriction or rapamycin led to an additional increase in LC3-II above that seen with c...Continue Reading

References

Apr 23, 2003·Experimental Cell Research·Ralph W KunclRobert Adams
Jun 21, 2007·The Journal of Biological Chemistry·Serhiy PankivTerje Johansen
Aug 23, 2007·Nature Reviews. Molecular Cell Biology·Daniel J Klionsky
Dec 23, 2008·Autophagy·Michael F N O'Leary, David A Hood
May 5, 2009·Autophagy·David C RubinszteinDaniel J Klionsky
Dec 17, 2009·The Journal of Cell Biology·Jeong-Sun JuConrad C Weihl
Feb 11, 2010·Cell·Noboru MizushimaBeth Levine

❮ Previous
Next ❯

Citations

Jun 15, 2011·Apoptosis : an International Journal on Programmed Cell Death·Bipul R AcharyaGopal Chakrabarti
Feb 15, 2013·Neurotherapeutics : the Journal of the American Society for Experimental NeuroTherapeutics·Conrad C Weihl
Mar 12, 2013·Acta Pharmacologica Sinica·Xiao-jie ZhangWei-dong Le
Nov 16, 2013·Cell Death & Disease·K CastilloC Hetz
Aug 17, 2012·American Journal of Physiology. Endocrinology and Metabolism·Jia QiuSrinivasan Dasarathy
Nov 29, 2015·The Journal of Physiology·Andreas M FritzenChristian Frøsig
Mar 5, 2016·The Journal of Physiological Sciences : JPS·Jeong-Sun JuJong-Moon Jeong
Jun 1, 2012·Journal of Cachexia, Sarcopenia and Muscle·Srinivasan Dasarathy
Mar 16, 2016·Cellular Signalling·Andreas Mæchel FritzenBente Kiens
Aug 5, 2015·Journal of Neuro-oncology·Kuan-Min FangShun-Fen Tzeng
Feb 12, 2014·Journal of Molecular and Cellular Cardiology·Changhan OuyangZhonglin Xie
Jul 23, 2013·Experimental Eye Research·Jonathan A WignesUsha P Andley
Apr 2, 2015·Anesthesiology·Ilan AzuelosBasil J Petrof
Oct 26, 2011·Autophagy·Andy BassDaniel J Klionsky
Nov 5, 2013·Autophagy·James K ChingConrad C Weihl
Apr 5, 2011·Autophagy·Jeffrey HaspelAugustine M K Choi
Sep 12, 2012·Autophagy·Daniel J KlionskyBrian Zuckerbraun
Aug 21, 2013·Autophagy·Mahroo MofarrahiSabah N A Hussain
Apr 4, 2015·Skeletal Muscle·Anna VainshteinDavid A Hood
Feb 12, 2013·The American Journal of Pathology·Fabio PennaPaola Costelli
Jan 26, 2012·Diabetes·Eijiro Yamada, Rajat Singh
Jul 28, 2016·Scientific Reports·Zaira AversaMaurizio Muscaritoli
Aug 16, 2016·The Journal of Clinical Investigation·Brian T O'NeillC Ronald Kahn
Sep 23, 2016·Autophagy·Silvia CastagnaroLuciano Merlini
Nov 8, 2016·Molecular Cell·Takeshi KaizukaNoboru Mizushima
Nov 23, 2016·PloS One·Haiming M LiuLaDora V Thompson
Jan 7, 2017·Autophagy·Federico PietrocolaGuido Kroemer
Jun 9, 2017·Cells·Manon Moulis, Cécile Vindis
Aug 29, 2017·International Journal of Molecular Sciences·Saori R Yoshii, Noboru Mizushima

❮ Previous
Next ❯

Related Concepts

Related Feeds

Autophagy & Model Organisms

Autophagy is a cellular process that allows degradation by the lysosome of cytoplasmic components such as proteins or organelles. Here is the latest research on autophagy & model organisms

Autophagy & Aging: Inhibitors

The feed focuses on the role of nuclear export inhibitors and their effect on autophagy and the aging process.