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Departments of 2 Medicine and 1 Cellular and Molecular Medicine, University of Ottawa and Ottawa General Hospital, Ottawa, Ontario, Canada K1H 8L6
Nitric oxide (NO)
derived from neuronal NO synthase (nNOS) in the kidney inner medulla
has been implicated in the regulation of arterial blood pressure. The
purpose of the present study was to determine the effect of high
dietary NaCl on the expression of nNOS in the rat inner medullary
collecting duct (IMCD). After 3 days or 3 wk of high (4.0%)-NaCl diet
in rats, urinary
NO
2/NO3 excretion significantly increased. In freshly microdissected IMCD, nNOS
was readily detected by immunofluorescence with polyclonal antibody, an
effect that was completely blocked by neutralization of antibody with
immunizing antigen. In rats fed a 4.0% NaCl diet for 3 days, IMCD nNOS
mRNA, detected by RT-PCR, did not change from control values (0.3%
NaCl, 19.84 ± 1.57 × 103, vs. 4.0% NaCl, 20.44 ± 3.14 × 103 cpm;
P = not significant,
n = 3). By Western blotting however, nNOS protein expression significantly increased (0.3% NaCl, 0.51 ± 0.12, vs. 4.0% NaCl, 0.92 ± 0.14 arbitrary units;
P < 0.05, n = 5). After 3 wk of 4.0% dietary
NaCl, expression of nNOS mRNA and protein in IMCD did not differ
significantly from control values. In contrast to these data, renal
cortical expression of nNOS mRNA and protein was significantly
decreased after 4.0% NaCl diet for 3 days. High dietary NaCl had no
significant effect on expression of mRNA for inducible NO synthase
(iNOS) in IMCD after either 3 days or 3 wk. In summary, our data
indicate that nNOS mRNA and protein are expressed in IMCD and that high
dietary NaCl differentially regulates nNOS expression in IMCD and
cortex. The early increase in nNOS protein in IMCD may contribute to
enhanced local production of NO and thereby represent an adaptive
response to salt intake.
nitric oxide; messenger ribonucleic acid expression; nitrates; inducible nitric oxide synthase
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