Endothelin-1 blunts transepithelial transport and differentiation of Madin-Darby canine kidney cells.

We investigated the effects of endothelin-1 (ET-1) on Madin-Darby canine kidney (MDCK) cells, a cell line originating from the renal collecting duct. The activity of transepithelial transport was assessed as the rate of dome formation in monolayers grown on solid support. The pH value of the dome fluid (dome pH) was measured by means of pH-selective microelectrodes. Differentiation of monolayer cells was estimated as the peanut-lectin(PNA)-binding capacity of the apical membrane. Confluent monolayers were incubated for 12-72 h in serum-free medium at various concentrations of ET-1. Exposure to 1 nmol/l ET-1 reduced dome formation by a maximum of 41 +/- 8% (n = 4; P less than 0.02) after 24 h. ET-1 (10 nmol/l; 24 h) decreased dome pH from 7.52 +/- 0.02 (n = 53) to 7.36 +/- 0.03 (n = 51; P less than 0.02). Apical application of amiloride (1 mmol/l) reduced dome pH in both ET-1-treated and non-treated domes to essentially the same level, 7.25 +/- 0.03 (n = 19) and 7.23 +/- 0.03 (n = 17) respectively. ET-1 (10 nmol/l; 24 h) reduced PNA-binding capacity by 19 +/- 3% (n = 5; P less than 0.02). Moreover, ET-1 prevented the increase in PNA binding (+ 53 +/- 7%; n = 5) induced by 0.1 mumol/l aldosterone. We conclude that ET-1 inhibits transepithelial transport and PNA binding via inhibition of apical Na+/H+ exchange, thus antagonizing aldosterone action in MDCK cells.