Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes

Abstract

Measuring the mitochondrial electron transfer complex (ETC) profile from previously frozen heart tissue samples from offspring born to an exercised sow provided descriptive data about exercise induced mitochondrial biochemical changes in heart tissue from the offspring born to the exercised sow. The hypothesis that was proposed and tested was that regular maternal exercise of a sow during pregnancy would increase the mitochondrial efficiency of offspring heart bioenergetics. This hypothesis was tested by isolating mitochondria using a mild-isolation procedure so that mitochondrial ETC, and supercomplex profiles were assessed. The procedure described here allowed for the processing of previously frozen archived heart tissues, and eliminated the necessity of fresh mitochondria preparation for the assessment of mitochondrial ETC complexes, supercomplexes, and ETC complex activity profiles. This protocol described the optimal ETC protein complex measurement in multiplexed antibody-based immunoblotting, and super complex assessment using blue-native gel electrophoresis.SUMMARYPreparation of mitochondria enriched samples from previously frozen archived solid tissues allowed the investigators to perform both functional and analytical assessments of mitochondria in various experimental modalities. This study demonstrated (i) how to prepare mitochondria enriched preparations from frozen heart tissue and (2) perform analytical assessments of mitochondria