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L-Arginine inhibits thick ascending limb (THAL) NaCl absorption by activating endothelial NO synthase (eNOS) and increasing NO production. Inhibition of renal NO production combined with a high-salt diet produces hypertension, and the THAL has been implicated in salt-sensitive hypertension. We hypothesized that a high-salt diet enhances the inhibitory action of L-arginine on NaCl absorption by THALs because of increased eNOS expression and NO production. To test this, we used isolated THALs from rats on a normal-salt (NS) or high-salt diet (HS) for 7 to 10 days. L-Arginine (1 mmol/L) decreased chloride absorption by 56+/-10% in THALs from rats on a HS diet, but only 29+/-3% in THALs from rats on a NS diet. eNOS expression in isolated THALs from rats on a HS diet was increased by 3.9-fold compared with NS (P&lt;0.03). However, L-arginine increased NO levels to the same extent in THALs from both groups, as measured with DAF-2 DA or a NO-sensitive electrode. To determine whether a HS diet increases the sensitivity of the THAL to NO, we tested the effects of the NO donor spermine NONOate on chloride absorption. In THALs from rats on a HS diet, 1 and 5 micromol/L spermine NONOate reduced chloride absorption by 35+/-5% and 58+/-6%, respectively. In contrast, these same concentrations of spermine NONOate reduced chloride absorption by 4+/-4% (P&lt;0.03 versus HS diet) and 43+/-9% in THALs from rats on a NS diet. We conclude that a HS diet enhances the effect of NO in the THAL. L-Arginine-stimulated NO production was not enhanced by a HS diet, despite increased eNOS protein.

High-Salt Diet Increases Sensitivity to NO and eNOS Expression But Not NO Production in THALs