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Am J Physiol Renal Physiol 279: F326-F333, 2000;
0363-6127/00 $5.00
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Vol. 279, Issue 2, F326-F333, August 2000

Endothelin inhibits thick ascending limb chloride flux via ETB receptor-mediated NO release

Craig F. Plato1, David M. Pollock2, and Jeffrey L. Garvin1

1 Division of Hypertension and Vascular Research, Henry Ford Hospital, Detroit, Michigan 48202; and 2 Vascular Biology Center, Medical College of Georgia, Augusta, Georgia 30912

Endothelin-1 (ET-1) inhibits transport in various nephron segments, and the thick ascending limb of the loop of Henle (TALH) expresses ET-1 receptors. In many tissues, activation of ETB receptors stimulates release of NO, and we recently reported that endogenous NO inhibits TALH chloride flux (JCl). However, the relationship between ET-1 and NO in the control of nephron transport has not been extensively studied. We hypothesized that ET-1 decreases NaCl transport by cortical TALHs through activation of ETB receptors and release of NO. Exogenous ET-1 (1 nM) decreased JCl from 118.3 ± 15.0 to 62.7 ± 13.6 pmol · mm-1 · min-1 (48.3 ± 8.2% reduction), whereas removal of ET-1 increased JCl in a separate group of tubules from 87.6 ± 10.7 to 115.2 ± 10.3 pmol · mm-1 · min-1 (34.5 ± 6.2% increase). To determine whether NO mediates the inhibitory effects of ET-1 on JCl, we examined the effect of inhibiting of NO synthase (NOS) with NG-nitro-L-arginine methyl ester (L-NAME) on ET-1-induced changes in JCl. L-NAME (5 mM) completely prevented the ET-1-induced reduction in JCl, whereas D-NAME did not. L-NAME alone had no effect on JCl. These data suggest that the effects of ET-1 are mediated by NO. Blockade of ETB receptors with BQ-788 prevented the inhibitory effects of 1 nM ET-1. Activation of ETB receptors with sarafotoxin S6c mimicked the inhibitory effect of ET-1 on JCl (from 120.7 ± 12.6 to 75.4 ± 13.3 pmol · mm-1 · min-1). In contrast, ETA receptor antagonism with BQ-610 did not prevent ET-1-mediated inhibition of TALH JCl (from 96.5 ± 10.4 to 69.5 ± 8.6 pmol · mm-1 · min-1). Endothelin increased intracellular calcium from 96.9 ± 14.0 to 191.4 ± 11.9 nM, an increase of 110.8 ± 26.1%. We conclude that exogenous endothelin indirectly decreases TALH JCl by activating ETB receptors, increasing intracellular calcium concentration, and stimulating NO release. These data suggest that endothelin acts as a physiological regulator of TALH NO synthesis, thus inhibiting chloride transport and contributing to the natriuretic effects of ET-1 observed in vivo.

kidney; nitric oxide; tubular transport


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