Modulation of Mg2+ influx and cytoplasmic free Mg2+ concentration in rat ventricular myocytes

To examine whether TRPM7, a member of the melastatin family of transient receptor potential channels, is a physiological pathway for Mg2+ entry in mammalian cells, we studied the effect of TRPM7 regulators on cytoplasmic free Mg2+ concentration ([Mg2+](i)) of rat ventricular myocytes. Acutely isolated single cells were AM-loaded with the fluorescent indicator furaptra, and [Mg2+](i) was estimated at 25 degrees C. After [Mg2+](i) was lowered by soaking the cells with a high-K+ and Mg2+-Ca2+-free solution, [Mg2+](i) was recovered by extracellular perfusion of Ca2+-free Tyrode's solution that contained 1mMMg(2+). The initial rate of increase in [Mg2+](i) was analyzed as the Mg2+ influx rate. The Mg2+ influx rate was increased by the TRPM7 activator, naltriben (2-50M), in a concentration-dependent manner with a half maximal effective concentration (EC50) of 24M. This EC50 value is similar to that reported for the activation of recombinant TRPM7 overexpressed in HEK293 cells. Naltriben (50M) caused little change in basal [Mg2+](i) (0.9mM) in Ca2+-free Tyrode's solution, but significantly raised [Mg2+](i) to 1.31 +/- 0.03mM in 94min after the removal of extracellular Na+. Re-introduction of extracellular Na+ lowered [Mg2+](i) back to the basal level even in the presence of naltriben. Application of 10M NS8593, an inhibitor of TRPM7, significantly lowered [Mg2+](i) to 0.72 +/- 0.03mM in 50-60min independent of extracellular Na+. The results suggest that Mg2+ entry through TRPM7 significantly contributes to physiological Mg2+ homeostasis in mammalian heart cells.