To study possible modulation of Mg²⁺ transport in low Mg²⁺ conditions, we fed either a Mg-deficient diet or a Mg-containing diet (control) to Wistar rats for 1–6 weeks. Total Mg concentrations in serum and cardiac ventricular tissues were measured by atomic absorption spectroscopy. Intracellular free Mg²⁺ concentration ([Mg²⁺]_i) of ventricular myocytes was measured with the fluorescent indicator furaptra. Mg²⁺ transport rates, rates of Mg²⁺ influx and Mg²⁺ efflux, were estimated from the rates of change in [Mg²⁺]_i during Mg loading/depletion and recovery procedures. In Mg-deficient rats, the serum total Mg concentration (0.29±0.026 mM) was significantly lower than in control rats (0.86±0.072 mM) after 4–6 weeks of Mg deficiency. However, neither total Mg concentration in ventricular tissues nor [Mg²⁺]_i of ventricular myocytes was significantly different between Mg-deficient rats and control rats. The rates of Mg²⁺ influx and efflux were not significantly different in both groups. In addition, quantitative RT-PCR revealed that Mg deficiency did not substantially change mRNA expression levels of known Mg²⁺ channels/transporters (TRPM6, TRPM7, MagT1, SLC41A1 and ACDP2) in heart and kidney tissues. These results suggest that [Mg²⁺]_i as well as the total Mg content of cardiac myocytes, was well maintained even under chronic hypomagnesemia without persistent modulation in function and expression of major Mg²⁺ channels/transporters in the heart.