Monitoring Mitochondrial Respiration in Mouse Cerebellar Granule Neurons

Ahmad Sharanek; Arezu Jahani-Asl

doi:10.1007/978-1-0716-2409-8_1

Monitoring Mitochondrial Respiration in Mouse Cerebellar Granule Neurons

Methods Mol Biol. 2022:2515:1-15. doi: 10.1007/978-1-0716-2409-8_1.

Authors

Ahmad Sharanek^{1

2

3}, Arezu Jahani-Asl^{4

5

6}

Affiliations

¹ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada.
² University of Ottawa Brain and Mind Research Institute, University of Ottawa, Ottawa, ON, Canada.
³ Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, QC, Canada.
⁴ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada. arezu.jahani@uottawa.ca.
⁵ University of Ottawa Brain and Mind Research Institute, University of Ottawa, Ottawa, ON, Canada. arezu.jahani@uottawa.ca.
⁶ Division of Experimental Medicine , McGill University, Montreal, QC, Canada. arezu.jahani@uottawa.ca.

PMID: 35776342
DOI: 10.1007/978-1-0716-2409-8_1

Abstract

Defects in mitochondrial oxidative phosphorylation have been observed in numerous neurodegenerative disorders and are linked to bioenergetic crises leading to neuronal death. The distinct metabolic profile of neurons is predominantly oxidative, which is characterized by the oxidation of glucose or its metabolites in the mitochondria to produce ATP. This process involves the tricarboxylic acid cycle, electron transfer in the respiratory chain, and oxygen consumption. Therefore, measurement of oxygen consumption rates (OCR) can be accurately applied to assess the rate of mitochondrial respiration. In this chapter, we describe our optimized protocol for the assessment of OCR specifically in primary mouse cerebellar granule neurons (CGN). The protocol includes isolation and manipulation of mouse CGNs followed by real-time assessment of mitochondrial OCR using a Seahorse XFe96 extracellular flux analyzer.

Keywords: Cerebellum; Mitochondrion; Oxidative phosphorylation; Primary neurons; Seahorse XFe96 extracellular flux analyzer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Respiration*
Mice
Mitochondria* / metabolism
Neurons / metabolism
Oxidative Phosphorylation
Respiration

Abstract

Publication types

MeSH terms

Grants and funding