Stable Isotope Tracer Analysis in Isolated Mitochondria from Mammalian Systems

Metabolites

Julie St-Pierre¹  Sylvia Andrzejewski¹  Daina Avizonis¹  Simon-Pierre Gravel¹ 

[1] Rosalind and Morris Goodman Cancer Research Centre, 1160 Pine Ave. West, Montréal H3A 1A3, Québec, Canada;

关键词: mitochondria;    muscle;    cells;    stable isotope tracer analysis;    citric acid cycle;    rotenone;    antimycin;    Neu/ErbB2;    cancer;    GC/MS;   

DOI  :  10.3390/metabo4020166

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown