In vivo autophagy quantification: Measuring LC3 and P62 puncta in 3D image system from zebrafish larvae.

Journal of Cellular Biochemistry
Abdalla Elbialy

Abstract

Autophagy is a central pathway in maintaining cellular homeostasis through the recycling of damaged proteins and organelles. Detection of LC3 protein levels by immunofluorescence or western blot analysis is one of the most common ways to measure autophagy. For quantitative autophagy analysis, LC3 western blot analysis is commonly used, whereas immunostaining is used for qualitative autophagy analysis. However, zebrafish larvae have a lot of proteases that rapidly degrade LC3 protein in samples. P62 is another autophagy marker that bind to damaged proteins and can reflects autophagic status. This study demonstrates a fast and accurate way to quantify autophagy from LC3 and/or P62 immunostaining images. We used a three-dimensional analysis of whole-mount LC3 immunostaining images of zebrafish larvae. Counting LC3 and P62 punctate by two dimensions can be used as a qualitative method for the analysis of autophagy. However, here we demonstrate that 3D image analysis can be used as a quantitative, rapid tool for monitoring autophagy in zebrafish larvae and avoiding drawbacks of LC3 western blot analysis.

References

Apr 8, 2004·Developmental Cell·Beth Levine, Daniel J Klionsky
Apr 23, 2004·Trends in Cell Biology·Ana Maria Cuervo
Aug 4, 2006·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Qing Jun WangZhenyu Yue
Aug 30, 2007·Proceedings of the National Academy of Sciences of the United States of America·Masaaki KomatsuZhenyu Yue
Apr 22, 2008·Methods in Molecular Biology·Isei TanidaEiki Kominami
Dec 1, 2009·Cell Metabolism·Eva MasieroMarco Sandri
Feb 11, 2010·Cell·Noboru MizushimaBeth Levine
Feb 9, 2012·World Journal of Biological Chemistry·Kadija AbounitRoy B McCauley
May 15, 2012·Biochemical and Biophysical Research Communications·Jianzhou CuiHan-Ming Shen
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Sep 12, 2012·Autophagy·Daniel J KlionskyBrian Zuckerbraun
Dec 9, 2014·Autophagy·Ben LoosJan-Hendrik S Hofmeyr
Dec 17, 2014·Methods : a Companion to Methods in Enzymology·Máté VargaTibor Vellai
Jan 8, 2016·Nature·Laura García-PratPura Muñoz-Cánoves
Oct 20, 2018·Biology·Abdalla ElbialyShigeharu Kinoshita
Oct 26, 2018·Open Biology·Ana LopezDavid C Rubinsztein
Aug 14, 2019·Zebrafish·Denhi SchnabelHilda Lomeli

❮ 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