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Am J Physiol Renal Physiol 275: F353-F360, 1998;
0363-6127/98 $5.00
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Vol. 275, Issue 3, F353-F360, September 1998

Mg2+/Ca2+ sensing inhibits hormone-stimulated Mg2+ uptake in mouse distal convoluted tubule cells

Brian W. Bapty, Long-Jun Dai, Gordon Ritchie, Lucie Canaff, Geoffrey N. Hendy, and Gary A. Quamme

Department of Medicine, University of British Columbia, University Hospital, Koerner Pavilion, Vancouver, British Columbia, Canada V6T 1Z3; and Departments of Medicine, Physiology and Human Genetics, McGill University and Royal Victoria Hospital, Montreal, Quebec, Canada H3A 1A1

The distal convoluted tubule plays a significant role in renal magnesium conservation. An immortalized mouse distal convoluted tubule (MDCT) cell line has been extensively used to study the cellular mechanisms of magnesium transport in this nephron segment. MDCT cells possess an extracellular polyvalent cation-sensing mechanism responsive to Mg2+, Ca2+, and neomycin. The present studies determined the effect of Mg2+/Ca2+ sensing on hormone-mediated cAMP formation and Mg2+ uptake in MDCT cells. MDCT cells were Mg2+ depleted by culturing in Mg2+-free media for 16 h, and Mg2+ uptake was measured by microfluorescence after placing the depleted cells in 1.5 mM MgCl2. The mean rate of Mg2+ uptake was 164 ± 5 nM/s in control MDCT cells. Activation of Mg2+/Ca2+ sensing with neomycin did not affect basal Mg2+ uptake (155 ± 5 nM/s). We have previously reported that treatment of MDCT cells with either glucagon or arginine vasopressin (AVP) stimulated Mg2+ entry. In the present studies, the addition of extracellular Mg2+ or Ca2+ inhibited glucagon- and AVP-stimulated cAMP formation and Mg2+ uptake in concentration-dependent manner with half-maximal concentrations of ~1.5 and 3.0 mM, respectively. Exogenous cAMP or forskolin stimulated Mg2+ uptake in the presence of Mg2+/Ca2+ sensing activation. We infer from these studies that Mg2+/Ca2+-sensing mechanisms located in the distal convoluted tubule may play a role in control of distal magnesium absorption.

intracellular magnesium; magnesium uptake; fluorescence; extracellular calcium; extracellular magnesium; neomycin; adenosine 3',5'-cyclic monophosphate measurements; glucagon; arginine vasopressin; neomycin


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