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Papers In Press, published online ahead of print September 11, 2000
NIDDK, NIH, Bethesda, MD 20892
Corresponding Author: jurgens{at}intra.niddk.nih.gov
Reducing luminal NaCl concentration in the macula densa region of the nephron stimulates renin secretion, and this response is blocked by a specific inhibitor of cyclooxygenase-2, COX-2 (1). To study whether low NaCl activates COX-2 activity or expression we clonally derived a macula densa cell line (MMDD1 cells) from SV-40 transgenic mice using FACS of renal tubular cells labelled with segment specific fluorescent lectins. MMDD1 cells express COX-2, bNOS, NKCC2, and ROMK, but not Tamm-Horsfall protein, and showed rapid 86Rb+ uptake that was inhibited by a reduction in NaCl concentration and by bumetanide or furosemide. Isosmotic exposure of MMDD1 cells to low NaCl (60 mM) caused a prompt and time-dependent stimulation of PGE2 release that was prevented by the COX-2 specific inhibitor NS-398 (10 µM). Reducing NaCl to 60 and 6 mM for 16 hours increased COX-2 expression in a chloride-dependent fashion. Low NaCl phosphorylated p38 kinase within 30 min and Erk1/2 kinases within 15 min without changing total MAP kinase levels. Low NaCl-stimulated PGE2 release and COX-2 expression was inhibited by SB 203580 and PD 98059 (10 µM), inhibitors of p38 and ERK kinase pathways. We conclude that low chloride stimulates PGE2 release and COX-2 expression in MMDD1 cells through activation of MAP kinases.
J. Biol. Chem, 10.1074/jbc.M006218200
Submitted on July 13, 2000
Revised on August 31, 2000
Accepted on September 9, 2000
Low chloride stimulation of PGE2 release and COX-2 expression in a mouse Macula densa cell line
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