Determination of epithelial magnesium transport with stable isotopes

The inability to adequately determine Mg(2+) flux rates with radiotracer studies has stymied our efforts to understand how magnesium is transported by epithelial cells. To evaluate epithelial Mg(2+) transport, a stable 25Mg isotope was used to measure magnesium uptake into normal and Mg(2+)-depleted Madin-Darby canine kidney (MDCK) cells. 25Mg entry rates were significantly increased in Mg(2+)-depleted cells relative to those cultured in normal magnesium media, 0.5 mM. 25Mg uptake was inhibited by external La(3+) but not Ca(2+) in both normal and Mg(2+)-depleted cells suggesting a specific entry pathway. These results with 25Mg were the same as with microfluorescence determinations using mag-fura-2. We have shown that Mg(2+) entry into epithelial cells reflects transepithelial transport; accordingly, increased Mg(2+) uptake in Mg(2+)-depleted cells provides an important intrinsic control of renal magnesium absorption. Furthermore, these studies indicate that cellular Mg(2+) transport may be quantitated with the use of stable isotopes that may be successfully applied to cells other than epithelia.