The mechanism(s1 of absorption of chloride ions (C1-absorption), as measured by the short-circuit current (I,,, pA-cm ' 1, was investigated in the intestinal middle part of the seawater-(SW) and the freshwater-(FW) adapted eel. Tissues, mounted in an Ussing chamber and continuously perfused by gravity flow on both sides with 20 mM HCOB -1% C 0 2 Ringer's solution at pH 8.0, developed a serosa-negative transepithelial voltage (V,, mV) and an I,, that were both lower in FW-adapted than in SW-adapted eels. Bilateral removal of Na+ or C1-from the bathing solutions reduced both electrical variables to zero. Removal of K + from or addition of Ba'' (1 mM) to the luminal bath markedly decreased I,, and V,. When a K'-free plus Ba2+-containing solution perfused the lumen, I,, and V, were almost completely abolished. Luminal bumetanide (10 MI, an inhibitor of Na+-K + -2C1-cotransport, reduced I,, and V, by about 90% in a few minutes. M) and DIDS (4,4'-diisothiocyanostilbene-2,2-disulphonic acid; 10 -4 M), inhibitors of Na+/H+ and C1-/HC03-exchangers, and hydrochlorothiazide (lop4 M), an inhibitor of the Na-Cl symport, had no effect on I,, and V, when added luminally. Serosal ouabain M) abolished I,, and V, in about 30 min. All the above experimental manoeuvres led to similar results in both SW-adapted and FW-adapted eels.The data suggest that in the intestine of both SW and FW eels, Cl--absorption is mediated by a luminally located neutral Na Amiloride (