Investigation of mitochondrial function represents an important parameter of cardiac physiology as mitochondria are involved in energy metabolism, oxidative stress, apoptosis, aging, mitochondrial @entity17903 and @entity137 . Given this, technologies to measure cardiac mitochondrial function are in demand. One technique that employs an integrative approach to measure mitochondrial function is respirometric oxidative phosphorylation (OXPHOS) analysis. The principle of respirometric OXPHOS assessment is centered around measuring @entity26 concentration utilizing a Clark electrode. As the permeabilized fiber bundle consumes @entity26 , @entity26 concentration in the closed chamber declines. Using selected substrate-inhibitor-uncoupler titration protocols, electrons are provided to specific sites of the electron transport chain, allowing evaluation of mitochondrial function. Prior to respirometric analysis of mitochondrial function, mechanical and chemical preparatory techniques are utilized to permeabilize the sarcolemma of muscle fibers. Chemical permeabilization employs @entity3857 to selectively perforate the cell membrane while maintaining cellular architecture. This paper thoroughly describes the steps involved in preparing @entity3857 -skinned cardiac fibers for @entity26 consumption measurements to evaluate mitochondrial OXPHOS. Additionally, troubleshooting advice as well as specific substrates, inhibitors and uncouplers that may be used to determine mitochondria function at specific sites of the electron transport chain are provided. Importantly, the described protocol may be easily applied to cardiac and skeletal tissue of various animal models and @entity1 samples.

Respirometric oxidative phosphorylation assessment in XXXX -permeabilized cardiac fibers.

@entity1 :: ('9606', 'Species') :: ['human'] @entity137 :: ('MESH:D064420', 'Disease') :: ['drug toxicity'] @entity26 :: ('MESH:D010100', 'Chemical') :: ['oxygen'] @entity3857 :: ('MESH:D012503', 'Chemical') :: ['saponin'] @entity17903 :: ('MESH:D017237', 'Disease') :: ['encephalomyopathies']
@entity3857 :: (MESH:D012503,Chemical) :: ['saponin']
