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Am J Physiol Renal Physiol 280: F815-F822, 2001;
0363-6127/01 $5.00
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Vol. 280, Issue 5, F815-F822, May 2001

The calcium-sensing receptor regulates calcium absorption in MDCK cells by inhibition of PMCA

Kristy A. Blankenship, J. Jason Williams, Martha S. Lawrence, Kenneth R. McLeish, William L. Dean, and John M. Arthur

Molecular Signaling Group and Departments of Medicine and Biochemistry and Molecular Biology, The University of Louisville, Louisville 40202 and Veterans Affairs Medical Center, Louisville, Kentucky 40206

Calcium transport across a monolayer of Madin-Darby canine kidney (MDCK) cells was measured in response to stimulation of the basal surface with calcium-sensing receptor (CaR) agonists. Stimulation of the CaR resulted in a time- and concentration-dependent inhibition of calcium transport but did not change transepithelial voltage or resistance. Inhibition of transport was not altered by pretreatment of cells with pertussis toxin but was blocked by the phospholipase C (PLC) inhibitor U-73122. To determine a potential mechanism by which the CaR could inhibit calcium transport, we measured activity of the plasma membrane calcium ATPase (PMCA). Stimulation of the CaR on the basal surface resulted in an inhibition of the PMCA in a concentration- and PLC-dependent manner. Thus stimulation of the CaR inhibits both calcium transport and PMCA activity through a PLC-dependent pathway. These studies provide the first direct evidence that calcium can inhibit its own transcellular absorption in a model of the distal tubule. In addition, they provide a potential mechanism for the CaR to inhibit calcium transport, inhibition of PMCA.

kidney tubules; distal; calcium-transporting adenosine 5'-triphosphatase; receptors; cell surface; kidney; ion transport; Madin-Darby canine kidney; plasma membrane calcium adenosine 5'-triphosphatase


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