Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes
Document Type
Article
Publication Title
Journal of Visualized Experiments: JoVE
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.
DOI
10.3791/62809
Publication Date
8-2021
Keywords
mitochondria, electron transport, electrons, energy metabolism, mitochondria, metabolism, oxidative phosphorylation, pregnancy, swine, heart
ISSN
1940-087X
Recommended Citation
Barrera D, Upton S, Rauch M, Notarianni T, Eum KS, Liberty M, Sah SV, Liu R, Newcomer S, May LE, Agbas E, Sage J, Agbas A. Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes. Journal of Visualized Experiments: JoVE. 2021; 174. doi: 10.3791/62809.
