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Abstract

Biochemistry

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

Published: August 16th, 2021

DOI:

10.3791/62809

1AdventHelath, 2University of Illinois Chicago School of Public Health, 3Lincoln Memorial University, 4Children’s Mercy Hospital, 5Scripps Clinic, 6Cincinnati Children’s Hospital Medical Center, 7Kansas City University, 8Roblex Rex Veterans Affairs Medical Center, 9California State University San Marcos, 10East Carolina University, 11Massachusetts Institute of Technology, 12Boehringer Ingelheim Norway KS, 13Heartland Center for Mitochondrial Medicine

Abstract

The mitochondrial electron transfer complex (ETC) profile is modified in the heart tissue of the offspring born to an exercised sow. The hypothesis proposed and tested was that a 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 to assess mitochondrial ETC and supercomplex profiles. 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 describes the optimal ETC protein complex measurement in multiplexed antibody-based immunoblotting and super complex assessment using blue-native gel electrophoresis.

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Keywords Mitochondrial Electron Transfer

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