Evaluation of basic mitochondrial functions using rat tissue homogenates

Mitochondrion. 2011 Sep;11(5):722-8. doi: 10.1016/j.mito.2011.05.006. Epub 2011 Jun 2.

Abstract

The primary attempt in diagnostic and experimental studies of numerous pathological states associated with mitochondrial dysfunction is a precise evaluation of changes in function, content and structure of mitochondrial OXPHOS system. The analysis of rat heart, liver, brain and kidney by oxygraphy, enzyme activities, membrane potential, and BN/SDS-PAGE western blotting demonstrated that tissue homogenates can substitute for isolated mitochondria, providing comparable qualitative mitochondrial parameters. The use of homogenate avoids the loss of the majority of mitochondria during their isolation. Only 50-100mg of the tissue is required for the complex OXPHOS analysis, i.e. five times less as compared with isolated mitochondria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / enzymology
  • Brain / metabolism*
  • Citrate (si)-Synthase / metabolism
  • Electron Transport Chain Complex Proteins / metabolism
  • Electron Transport Complex IV / metabolism
  • Enzyme Assays
  • Kidney / enzymology
  • Kidney / metabolism*
  • Liver / enzymology
  • Liver / metabolism*
  • Male
  • Membrane Potential, Mitochondrial
  • Mitochondria / enzymology
  • Mitochondria / metabolism*
  • Mitochondrial Proteins / metabolism
  • Myocardium / enzymology
  • Myocardium / metabolism*
  • Oxidative Phosphorylation
  • Oxygen Consumption
  • Rats
  • Rats, Wistar

Substances

  • Electron Transport Chain Complex Proteins
  • Mitochondrial Proteins
  • Electron Transport Complex IV
  • Citrate (si)-Synthase