Background: Sphingosine‐1‐phosphate plays vital roles in cardiomyocyte physiology, myocardial ischemia–reperfusion injury, and ischemic preconditioning. The function of the cardiomyocyte sphingosine‐1‐phosphate receptor 1 (S1P₁) in vivo is unknown.

Methods and Results: Cardiomyocyte‐restricted deletion of S1P₁ in mice (S1P₁^αMHCCre) resulted in progressive cardiomyopathy, compromised response to dobutamine, and premature death. Isolated cardiomyocytes from S1P₁^αMHCCre mice revealed reduced diastolic and systolic Ca²⁺ concentrations that were secondary to reduced intracellular Na⁺ and caused by suppressed activity of the sarcolemmal Na⁺/H⁺ exchanger NHE‐1 in the absence of S1P₁. This scenario was successfully reproduced in wild‐type cardiomyocytes by pharmacological inhibition of S1P₁ or sphingosine kinases. Furthermore, Sarcomere shortening of S1P₁^αMHCCre cardiomyocytes was intact, but sarcomere relaxation was attenuated and Ca²⁺ sensitivity increased, respectively. This went along with reduced phosphorylation of regulatory myofilament proteins such as myosin light chain 2, myosin‐binding protein C, and troponin I. In addition, S1P₁ mediated the inhibitory effect of exogenous sphingosine‐1‐phosphate on β‐adrenergic–induced cardiomyocyte contractility by inhibiting the adenylate cyclase. Furthermore, ischemic precondtioning was abolished in S1P₁^αMHCCre mice and was accompanied by defective Akt activation during preconditioning.

Conclusions: Tonic S1P₁ signaling by endogenous sphingosine‐1‐phosphate contributes to intracellular Ca²⁺ homeostasis by maintaining basal NHE‐1 activity and controls simultaneously myofibril Ca²⁺ sensitivity through its inhibitory effect on adenylate cyclase. Cardioprotection by ischemic precondtioning depends on intact S1P₁ signaling. These key findings on S1P₁ functions in cardiac physiology may offer novel therapeutic approaches to cardiac diseases.