title: Insulin stimulates Mg2+ uptake in mouse distal convoluted tubule cells.
abstract: Insulin has been shown to be a magnesium-conserving hormone acting, in part, through stimulation of magnesium absorption within the thick ascending limb. Although the distal convoluted tubule possesses the most insulin receptors, it is unclear what, if any, actions insulin has in the distal tubule. The effects of insulin were studied on immortalized mouse distal convoluted tubule (MDCT) cells by measuring cellular cAMP formation with radioimmunoassays and Mg2+ uptake with fluorescence techniques using mag-fura 2. To assess Mg2+ uptake, MDCT cells were first Mg(2+) depleted to 0.22 +/- 0.01 mM by culturing in Mg2+-free media for 16 h and then placed in 1.5 mM MgCl2, and the changes in intracellular Mg2+ concentration ([Mg2+]i) were measured with microfluorescence. [Mg2+]i returned to basal levels, 0.53 +/- 0.02 mM, with a mean refill rate, d([Mg2+]i)/dt, of 164 +/- 5 nM/s. Insulin stimulated Mg2+ entry in a concentration-dependent manner with maximal response of 214 +/- 12 nM/s, which represented a 30 +/- 5% increase in the mean uptake rate above control values. This was associated with a 2.5-fold increase in insulin-mediated cAMP generation (52 +/- 3 pmol. mg protein(-1). 5 min(-1)). Genistein, a tyrosine kinase inhibitor, diminished insulin-stimulated Mg2+ uptake (169 +/- 11 nM/s), but did not change insulin-mediated cAMP formation (47 +/- 5 pmol. mg protein(-1). 5 min(-1)). PTH stimulates Mg2+ entry, in part, through increases in cAMP formation. Insulin and PTH increase Mg2+ uptake in an additive fashion. In conclusion, insulin mediates Mg2+ entry, in part, by a genistein-sensitive mechanism and by modifying hormone-responsive transport. These studies demonstrate that insulin stimulates Mg2+ uptake in MDCT cells and suggest that insulin acts in concert with other peptide and steroid hormones to control magnesium conservation in the distal convoluted tubule.
authors.affiliation: Department of Medicine, University of British Columbia, Vancouver Hospital and Health Sciences Centre, Koerner Pavilion, Vancouver, British Columbia, Canada V6T 1Z3.
authors.forename: L J
authors.initials: LJ
authors.lastname: Dai
authors.forename: G
authors.initials: G
authors.lastname: Ritchie
authors.forename: B W
authors.initials: BW
authors.lastname: Bapty
authors.forename: D
authors.initials: D
authors.lastname: Kerstan
authors.forename: G A
authors.initials: GA
authors.lastname: Quamme
mesh_terms: D000450:Aldosterone / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D019833:Genistein / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology*; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D009355:Neomycin / Q000494:pharmacology; D010281:Parathyroid Hormone / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D013873:Thionucleotides / Q000494:pharmacology
chemical_list: D007328:Insulin; D010281:Parathyroid Hormone; D013873:Thionucleotides; C016957:adenosine-3',5'-cyclic phosphorothioate; D000450:Aldosterone; D019833:Genistein; D000242:Cyclic AMP; D011505:Protein-Tyrosine Kinases; D009355:Neomycin; D008274:Magnesium