To demonstrate that osmotic work can be accomplished across the inner medullary collecting duct (IMCD) by the difference in reflection coefficients for urea and NaCl, phenomenological coefficients for urea and NaCl transport were determined in isolated segments of the hamster IMCD perfused in vitro. Arginine vasopressin at 100 microU/ml increased urea permeability from 11.5 +/- 2.9 to 31.7 +/- 4.2 x 10(-7) cm2 s-1 in the middle IMCD but not in the upper IMCD. Urea transport in the middle IMCD consisted of two components, transport with saturable kinetics and simple passive diffusion. Permeability to Na+ was very low (2 x 10(-7) cm2 s-1). Reflection coefficients as measured by the equiosmolality method, with raffinose being a reference solute, were 0.87 +/- 0.05 and 0.71 +/- 0.04 for urea and 1.03 +/- 0.07 and 0.91 +/- 0.04 for NaCl in the upper and the middle IMCD, respectively. Reflection coefficient for urea in the middle IMCD was 0.68 when determined by the zero volume flux method. When the middle IMCD was perfused with bicarbonate Krebs-Ringer (BKR) solution containing 200 mmol/l urea, the replacement of urea in the bathing fluid with equisomolal NaCl caused large volume flux (3.81 +/- 0.45 nl mm-1 min-1) associated with dilatation of intercellular space. The existence of vasopressin in the bath was essential for this phenomenon. This effect was inhibited by 5 x 10(-4) M phloretin in the bath, suggesting that the vasoressin-stimulated urea transport is responsible for this phenomenon. From these observations, we conclude that transport parameters of the middle IMCD are appropriate for accomplishment of osmotic work across this segment in the absence of physicochemical osmotic gradients.