Altered calcium handling in cardiomyocytes from arginine:glycine amidinotransferase knockout mice is rescued by creatine.
Laasmaa M., Branovets J., Barsunova K., Karro N., Lygate CA., Birkedal R., Vendelin M.
The creatine kinase system facilitates energy transfer between mitochondria and the major ATPases in the heart. Creatine-deficient mice, which lack arginine:glycine amidinotransferase (AGAT) to synthesize creatine and homoarginine, exhibit reduced cardiac contractility. We studied how the absence of a functional CK system influences calcium handling in isolated cardiomyocytes from AGAT knockouts and wild-type littermates as well as in AGAT knockout mice receiving lifelong creatine supplementation via the food. Using a combination of whole-cell patch clamp and fluorescence microscopy, we demonstrate that the L-type calcium channel (LTCC) current amplitude and voltage range of activation was significantly lower in AGAT knockout compared to wild-type littermates. Additionally, the inactivation of LTCC and the calcium transient decay were significantly slower. According to our modeling results, these changes can be reproduced by reducing three parameters in knockout mice when compared to wild-type: LTCC conductance, the exchange constant of calcium transfer between subspace and cytosol, and SERCA activity. Since tissue expression of LTCC and SERCA protein were not significantly different between genotypes, this suggests the involvement of post-translational regulatory mechanisms or structural reorganization. The AGAT knockout phenotype of calcium handling was fully reversed by dietary creatine supplementation throughout life. Our results indicate reduced calcium cycling in cardiomyocytes from AGAT knockouts and suggest that the creatine kinase system is important for the development of calcium handling in the heart.