Mitochondrial respiration and ROS emission during β-oxidation in the heart: An experimental-computational study
Fig 4
Experimental and computational redox behavior of mitochondrial NAD(P)H in the presence of PCoA-driven respiration, and its impact on mitochondrial function.
Freshly isolated heart mitochondria from Sham or T1DM GPs were assayed in a fluorimeter in the presence of PCoA at the indicated concentrations, and as described in detail under Materials and Methods. (A) The first arrow in panel A indicates the addition time of mitochondria and 0.5mM malate (to enable the regeneration of the mitochondrial coenzyme A pool), after β-oxidation is triggered with the addition of both PCoA and L-carnitine (second arrow). Subsequent sequential additions of 5mM each of G/M and 1mM ADP were performed to test the state 4→3 transition. Depicted are the results from simultaneous monitoring of NAD(P)H in a typical experiment performed with Sham mitochondria but similar results were observed in mitochondria from diabetic GPs. (B) Model simulations of the transient, time-dependent, behavior of NADH upon successive addition of PCoA followed by glutamate and ADP to test the state 4→3 transition as indicated. The simulations condition mimic the experimental protocol in order to reproduce the experimental time courses depicted in panel A. All other model parameters are the same to those indicated in S1 Text.