Mitochondrial Matrix K+ Flux Independent of Large-conductance Ca2+-activated K+ Channel Opening
Format of Original
American Physiological Society
American Journal of Physiology - Cell Physiology
Original Item ID
Large-conductance Ca2+-activated K+ channels (BKCa) in the inner mitochondrial membrane may play a role in protecting against cardiac ischemia-reperfusion injury. NS1619 (30 μM), an activator of BKCa channels, was shown to increase respiration and to stimulate reactive oxygen species generation in isolated cardiac mitochondria energized with succinate. Here, we tested effects of NS1619 to alter matrix K+, H+, and swelling in mitochondria isolated from guinea pig hearts. We found that 30 μM NS1619 did not change matrix K+, H+, and swelling, but that 50 and 100 μM NS1619 caused a concentration-dependent increase in matrix K+ influx (PBFI fluorescence) only when quinine was present to block K+/H+ exchange (KHE); this was accompanied by increased mitochondrial matrix volume (light scattering). Matrix pH (BCECF fluorescence) was decreased slightly by 50 and 100 μM NS1619 but markedly more so when quinine was present. NS1619 (100 μM) caused a significant leak in lipid bilayers, and this was enhanced in the presence of quinine. The K+ ionophore valinomycin (0.25 nM), which like NS1619 increased matrix volume and increased K+ influx in the presence of quinine, caused matrix alkalinization followed by acidification when quinine was absent, and only alkalinization when quinine was present. If K+ is exchanged instantly by H+ through activated KHE, then matrix K+ influx should stimulate H+ influx through KHE and cause matrix acidification. Our results indicate that KHE is not activated immediately by NS1619-induced K+ influx that NS1619 induces matrix K+ and H+ influx through a nonspecific transport mechanism, and that enhancement with quinine is not due to the blocking of KHE, but to a nonspecific effect of quinine to enhance current leak by NS1619.
Aldakkak, Mohammed; Stowe, David F.; Cheng, Qunli; Kwok, Wai-Meng; and Camara, Amadou K.S., "Mitochondrial Matrix K+ Flux Independent of Large-conductance Ca2+-activated K+ Channel Opening" (2010). Biomedical Engineering Faculty Research and Publications. 336.