Analyzing autophagosomes and mitophagosomes in the mouse brain using electron microscopy

Kaizheng Duan; Ronald S Petralia; Ya-Xian Wang; Zheng Li

doi:10.1016/j.xpro.2022.101154

Analyzing autophagosomes and mitophagosomes in the mouse brain using electron microscopy

STAR Protoc. 2022 Feb 3;3(1):101154. doi: 10.1016/j.xpro.2022.101154. eCollection 2022 Mar 18.

Authors

Kaizheng Duan¹, Ronald S Petralia², Ya-Xian Wang², Zheng Li¹

Affiliations

¹ Section on Synapse Development Plasticity, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892, USA.
² Advanced Imaging Core, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, MD 20892, USA.

Abstract

Electron microscopy (EM) is considered the gold standard for studying macroautophagy and mitophagy, essential cellular processes for brain health. Here, we present a protocol using EM to analyze autophagosomes and mitophagosomes in the mouse amygdala. We describe the preparation of brain sections, followed by staining and EM imaging. We then detail the steps to identify and analyze autophagosome-like and mitophagosome-like structures. This protocol can be easily adapted to analyze autophagosomes and mitophagosomes in other mouse brain regions. For complete details on the use and execution of this protocol, please refer to Duan et al. (2021).

Keywords: Cell Biology; Microscopy; Neuroscience.

Publication types

Research Support, N.I.H., Intramural

MeSH terms

Animals
Autophagosomes*
Brain / diagnostic imaging
Mice
Microscopy, Electron
Mitophagy*
Staining and Labeling

Grants and funding

ZIA MH002881/ImNIH/Intramural NIH HHS/United States