Inositol 1,4,5-trisphosphate-mediated sarcoplasmic reticulum–mitochondrial crosstalk influences adenosine triphosphate production via mitochondrial Ca2+ uptake through the mitochondrial ryanodine receptor in cardiac myocytes

Elevated levels of inositol 1,4,5-trisphosphate (IP3) in adult cardiac myocytes are typically associated with the development of cardiac hypertrophy, arrhythmias, and heart failure. IP3 enhances intracellular Ca2+ release via IP3 receptors (IP(3)Rs) located at the sarcoplasmic reticulum (SR). We aimed to determine whether IP3-induced Ca2+ release affects mitochondrial function and determine the underlying mechanisms. We compared the effects of IP(3)Rs- and ryanodine receptors (RyRs)-mediated cytosolic Ca2+ elevation achieved by endothelin-1 (ET-1) and isoproterenol (ISO) stimulation, respectively, on mitochondrial Ca2+ uptake and adenosine triphosphate (ATP) generation. Both ET-1 and isoproterenol induced an increase in mitochondrial Ca2+ (Ca-m(2 +)) but only ET-1 led to an increase in ATP concentration. ET-1-induced effects were prevented by cell treatment with the IP3 antagonist 2-aminoethoxydiphenyl borate and absent in myocytes from transgenic mice expressing an IP3 chelating protein (IP3 sponge). Furthermore, ET-1-induced mitochondrial Ca2+ uptake was insensitive to the mitochondrial Ca2+ uniporter inhibitor Ru360, however was attenuated by RyRs type 1 inhibitor dantrolene. Using real-time polymerase chain reaction, we detected the presence of all three isoforms of IP(3)Rs and RyRs in murine ventricular myocytes with a dominant presence of type 2 isoform for both receptors. Stimulation of IP(3)Rs with ET-1 induces Ca2+ release from the SR which is tunnelled to mitochondria via mitochondrial RyR leading to stimulation of mitochondrial ATP production.