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Autophagy pp 529-534 | Cite as

Methods to Image Macroautophagy in the Brain In Vivo

  • Xigui Chen
  • Kanoh Kondo
  • Hitoshi OkazawaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1880)

Abstract

Macroautophagy is the process to remove intracellular organelles or proteins by using autophagosome that is composed of autophagy proteins such as atg3, atg7, and atg8/LC3 (Mizushima, et al. Annu Rev Cell Dev Biol. 27:107–132, 2011). Here, we develop a useful method for in vivo imaging of autophagosome under the two-photon microscopy. Time-lapse imaging of LC3-ECFP enables us to quantify the dynamics of number, size, and signal intensity of autophagosomes in neurons or in other types of cells in the brain.

Key words

Macroautophagy Autophagosome Two-photon microscopy 

Notes

Acknowledgments

This work was supported by “Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS),” “Strategic Research Program for Brain Sciences (SRPBS),” and a Grant-in-Aid for Scientific Research on Innovative Areas (Foundation of Synapse and Neurocircuit Pathology) from the Ministry of Education, Culture, Sports, Science and Technology of Japan. We also thank Drs. Kazuhiko Tagawa and Kyota Fujita for supporting ethics approval, technical development, and discussion.

References

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    Mizushima N, Yoshimori T, Ohsumi Y (2011) The role of Atg proteins in autophagosome formation. Annu Rev Cell Dev Biol 27:107–132CrossRefGoogle Scholar
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    Chen X, Kondo K, Motoki K, Homma H, Okazawa H (2015) Fasting activates macroautophagy in neurons of Alzheimer's disease mouse model but is insufficient to degrade amyloid-beta. Sci Rep 5:12115CrossRefGoogle Scholar
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    Sato M, Kawano M, Yanagawa Y, Hayashi Y (2016) In vivo two-photon imaging of striatal neuronal circuits in mice. Neurobiol Learn Mem 135:146–151CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Neuropathology, Medical Research Institute and Center for Brain Integration ResearchTokyo Medical and Dental UniversityTokyoJapan

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