Metabolites 2014, 4(2), 166-183; doi:10.3390/metabo4020166
Article

Stable Isotope Tracer Analysis in Isolated Mitochondria from Mammalian Systems

1,2,†email, 1,2,†email, 1email and 1,2,* email
Received: 21 November 2013; in revised form: 26 February 2014 / Accepted: 24 March 2014 / Published: 10 April 2014
(This article belongs to the Special Issue Cancer Metabolomics)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: Mitochondria are a focal point in metabolism, given that they play fundamental roles in catabolic, as well as anabolic reactions. Alterations in mitochondrial functions are often studied in whole cells, and metabolomics experiments using 13C-labeled substrates, coupled with mass isotopomer distribution analyses, represent a powerful approach to study global changes in cellular metabolic activities. However, little is known regarding the assessment of metabolic activities in isolated mitochondria using this technology. Studies on isolated mitochondria permit the evaluation of whether changes in cellular metabolic activities are due to modifications in the intrinsic properties of the mitochondria. Here, we present a streamlined approach to accurately determine 13C, as well as 12C enrichments in isolated mitochondria from mammalian tissues or cultured cells by GC/MS. We demonstrate the relevance of this experimental approach by assessing the effects of drugs perturbing mitochondrial functions on the mass isotopomer enrichment of metabolic intermediates. Furthermore, we investigate 13C and 12C enrichments in mitochondria isolated from cancer cells given the emerging role of metabolic alterations in supporting tumor growth. This original method will provide a very sensitive tool to perform metabolomics studies on isolated mitochondria.
Keywords: mitochondria; muscle; cells; stable isotope tracer analysis; citric acid cycle; rotenone; antimycin; Neu/ErbB2; cancer; GC/MS
PDF Full-text Download PDF Full-Text [495 KB, uploaded 10 April 2014 13:20 CEST]

Export to BibTeX |
EndNote


MDPI and ACS Style

Gravel, S.-P.; Andrzejewski, S.; Avizonis, D.; St-Pierre, J. Stable Isotope Tracer Analysis in Isolated Mitochondria from Mammalian Systems. Metabolites 2014, 4, 166-183.

AMA Style

Gravel S-P, Andrzejewski S, Avizonis D, St-Pierre J. Stable Isotope Tracer Analysis in Isolated Mitochondria from Mammalian Systems. Metabolites. 2014; 4(2):166-183.

Chicago/Turabian Style

Gravel, Simon-Pierre; Andrzejewski, Sylvia; Avizonis, Daina; St-Pierre, Julie. 2014. "Stable Isotope Tracer Analysis in Isolated Mitochondria from Mammalian Systems." Metabolites 4, no. 2: 166-183.

Metabolites EISSN 2218-1989 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert