Nitric Oxide Decreases the Permselectivity of the Paracellular Pathway in Thick Ascending Limbs

Abstract

Thick ascending limbs reabsorb 25% to 30% of the filtered NaCl. About 50% to 70% is reabsorbed via the transcellular pathway and 30% to 50% is reabsorbed through the Na-selective paracellular pathway. Nitric oxide (NO) inhibits transepithelial Na reabsorption, but its effects on the paracellular pathway are unknown. We hypothesized that NO decreases the selectivity of the paracellular pathway in thick ascending limbs via cGMP-dependent protein kinase. To assess relative Na/Cl permeability ratios (P Na /P Cl), we perfused rat thick ascending limbs and measured the effect of reducing bath NaCl on transepithelial voltage, creating dilution potentials, with vehicle, NO donors, and endogenous NO. P Na /P Cl was calculated using the Goldman-Hodgkin-Katz equation. Reducing bath Na/Cl to 16/8, 32/24, and 64/56 mmol/L created dilution potentials of -13.6±2.2, -10.8±3.0, and -6.1±0.9 mV, respectively. Calculated P Na /P Cl s were 2.0±0.2, 2.2±0.5, and 1.9±0.2. The NO donor spermine NONOate (200 mol/L) blunted the dilution potential caused by 32/24 mmol/L Na/Cl from -11.1±2.1 to -6.5±1.6 mV (P<0.004) and P Na /P Cl from 2.2±0.4 to 1.5±0.2. Nitroglycerin (200 μmol/L), another NO donor, also reduced P Na /P Cl. Controls showed no significant changes. Dibutyryl-cGMP decreased dilution potentials from -13.4±2.9 to -7.5±1.8 mV (n=6; P<0.01). cGMP-dependent protein kinase inhibition with KT5823 (4 μmol/L) blocked the effect of spermine NONOate, whereas phosphodiesterase 2 inhibition did not. Endogenously produced NO mimicked the effect of the NO donors. In conclusion, NO reduces the selectivity of the paracellular pathway in thick ascending limbs via cGMP and cGMP-dependent protein kinase.