The mitochondrial bioenergetic phenotype for protection from cardiac ischemia in SUR2 mutant mice

Abstract

The sulfonylurea receptor-2 (SUR2) is a subunit of ATP-sensitive potassium channels (KATP) in heart. Mice with the SUR2 gene disrupted (SUR2m) are constitutively protected from ischemia-reperfusion (I/R) cardiac injury. This was surprising because KATP, either sarcolemmal or mitochondrial or both, are thought to be important for cardioprotection. We hypothesized that SUR2m mice have an altered mitochondrial phenotype that protects against I/R. Mitochondrial membrane potential (ΔΨm), tolerance to Ca2+ load, and reactive oxygen species (ROS) generation were studied by fluorescence-based assays, and volumetric changes in response to K+ were measured by light scattering in isolated mitochondria. For resting SUR2m mitochondria compared with wild type, the ΔΨm was less polarized (46.1 ± 0.4 vs. 51.9 ± 0.6%), tolerance to Ca2+ loading was increased (163 ± 2 vs. 116 ± 2 μM), and ROS generation was enhanced with complex I [8.5 ± 1.2 vs. 4.9 ± 0.2 arbitrary fluorescence units (afu)/s] or complex II (351 ± 51.3 vs. 166 ± 36.2 afu/s) substrates. SUR2m mitochondria had greater swelling in K+ medium (30.2 ± 3.1%) compared with wild type (14.5 ± 0.6%), indicating greater K+ influx. Additionally, ΔΨm decreased and swelling increased in the absence of ATP in SUR2m, but the sensitivity to ATP was less compared with wild type. When the mitochondria were subjected to hypoxia-reoxygenation, the decrease in respiration rates and respiratory control index was less in SUR2m. ΔΨm maintenance in the SUR2m intact myocytes was also more tolerant to metabolic inhibition. In conclusion, the cardioprotection observed in the SUR2m mice is associated with a protected mitochondrial phenotype resulting from enhanced K+ conductance that partially dissipated ΔΨm. These results have implications for possible SUR2 participation in mitochondrial KATP.