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J. Biol. Chem., Vol. 275, Issue 48, 37922-37929, December 1, 2000
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From the 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) (Traynor, T. R., Smart, A., Briggs, J. P., and Schnermann, J. (1999) Am. J. Physiol. Renal Physiol. 277, F706-710). 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 fluorescence-activated cell
sorting of renal tubular cells labeled 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 prostaglandin
E2 (PGE2) release that was prevented by the
COX-2 specific inhibitor NS-398 (10 µM). Reducing NaCl to
60 and 6 mM for 16 h 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.
Low Chloride Stimulation of Prostaglandin E2
Release and Cyclooxygenase-2 Expression in a Mouse Macula Densa Cell
Line*
,,,,
,,, and**
NIDDK and NHLBI, National Institutes
of Health, Bethesda, Maryland 20892, the § Department of
Urology, University of Michigan, Ann Arbor, Michigan 48109, and the
¶ Department of Pathology and Laboratory Medicine, University of
Rochester, Rochester, New York 14642
*
This work was supported by NIDDK, National Institutes of
Health Grants DK-37448, DK-39255, and DK-40042 and by intramural funds
from NIDDK and NHLBI.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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