Key result
Maternal exercise during pregnancy resulted in a non-significant increase in Complex I-II activity in offspring cardiac tissue compared to sedentary controls (P>0.05).
Why the study?
Regular maternal exercise during pregnancy may increase mitochondrial efficiency in offspring heart bioenergetics, but methods to assess this from archived frozen tissue were needed.
This protocol demonstrates a viable method for isolating functional mitochondria from previously frozen archived solid heart tissues for biochemical assessment.
Non-significant effect on offspring cardiac mitochondrial activity in this animal model; leaves open translational relevance while validating mitochondrial isolation from archived tissue.
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 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. SUMMARY Preparation 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 demonstrates how to prepare mitochondria-enriched preparations from frozen heart tissue and perform analytical assessments of mitochondria.
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Barrera et al. (2021) studied Mitochondrial bioenergetics in cardiac tissue. Maternal aerobic exercise during pregnancy vs. Sedentary pregnancy was evaluated on Complex I-II activity (p=>0.05). Maternal exercise during pregnancy resulted in a non-significant increase in Complex I-II activity in offspring cardiac tissue compared to sedentary controls (P>0.05).
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