Luminal and basolateral endothelin inhibit chloride reabsorption in the mouse thick ascending limb via a...

1. The recent localization of endothelin synthesis and receptors in the thick ascending limb (TAL) prompted us to investigate a possible autocrine and/or paracrine effect of this agent. The net chloride flux (JCl) has been determined in isolated cortical and medullary TALs by the in vitro microperfusion technique. 2. In both segments, endothelin 1 (ET-1) at 10(-8) M in the bath significantly decreased JCl, an effect which was partially reversible and observed at concentrations equal to or greater than 10(-13) M. 3. This JCl inhibition (by 33.9 +/- 3.2%) was blocked by BQ788 and was also observed with sarafotoxin 6C and ET-3, indicating that endothelin receptor B (ETB) are present in TAL. 4. ET-1 did not affect cAMP content under basal or hormone-stimulated conditions. The presence of a prostaglandin synthesis inhibitor also did not prevent the ET-1 action on JCl. 5. The ET-1-induced inhibition of JCl was prevented by protein kinase C inhibitors (staurosporine or GF 109203) and was reproduced by diacylglycerol analogues (OAG and DiC8). However, ET-1 failed to increase intracellular Ca2+ concentration. 6. Addition of ET-1 or ET-3 to the apical surface induced a decrease of JCl throgh ETB receptors, an effect which was not additive with that induced by basolateral ET-1, and was not concomitant with an increase in intracellular Ca2+ concentration. 7. It is concluded that the basolateral and luminal inhibitions of JCl by ET-1 in TAL, through ETB receptors, is mediated by a protein kinase C activation which is independent of intracellular Ca2+ increase.