Calcium and Bicarbonate Signaling Pathways have Pivotal, Resonating Roles in Matching ATP Production to Demand

Mitochondrial ATP production in cardiac ventricular myocytes must be continually adjusted to rapidly replenish the ATP consumed by the working heart. Two systems are known to be critical in this regulation: mitochondrial matrix Ca²⁺([Ca²⁺]_m) and blood flow that is tuned by local ventricular myocyte metabolic signaling. However, these two regulatory systems do not account for the large physiological range of ATP consumption observed. We report here on the identity, location, and signaling cascade of a controversial third regulatory system -- CO₂/bicarbonate. CO₂is generated in the mitochondrial matrix as a metabolic waste product produced by oxidation of nutrients which power the production of ATP. It is a lipid soluble gas that equilibrates with bicarbonate (HCO₃−) in aqueous solutions. The bicarbonate level is tracked by a bicarbonate-activated adenylyl cyclase, soluble adenylyl cyclase (sAC). Using structural Airyscan super-resolution imaging and functional measurements we find that sAC is primarily inside the mitochondria of ventricular myocytes where it generates cAMP when activated by HCO₃−. This cAMP signaling cascade is shown to operate inside the mitochondrial inter-membrane space (IMS) by activating local EPAC1 (ExchangeProtein directlyActivated bycAMP) which turns on Rap1 (Ras-related protein 1). Thus, mitochondrial ATP production is shown to be increased by bicarbonate-triggered sAC signaling through Rap1. Additional evidence is presented indicating that the cAMP signaling itself does not occur directly in the matrix. We also show that this third signaling process involving bicarbonate and sAC activates the cardiac mitochondrial ATP production machinery by working independently of, yet in conjunction with, [Ca²⁺]_m-dependent ATP production to meet the energy needs of cellular activity in both health and disease. Thus, the resonant, or complementary effects of bicarbonate and [Ca²⁺]_msignaling arms tune mitochondrial ATP production to match the full scale of energy consumption in cardiac ventricular myocytes.