The mode and mechanism of phosphate transport in the light portion of the rabbit cortical collecting duct were studied with isolated tubular perfusion. Control studies (n = 48) revealed a mean transepithelial potential difference (PD) of -16.5 +/- 1.3 mV and net phosphate absorption (JPO4) of 0.58 +/- 0.07 pmol.mm-1.min-1. To characterize transport, the effects of flow rate, ouabain, amiloride, and phosphate concentration, and parathyroid hormone (PTH) were analyzed. There was a significant linear relationship between JPO4 and flow rate (n = 48) (r = 0.71, P less than 0.025). Increasing the difference between lumen and bath phosphate concentration from -8.0 to +8.0 mg/dl was associated with a stepwise increase in JPO4 from -0.18 +/- 0.02 to 0.93 +/- 0.05 pmol.mm-1.min-1. PTH had no effect on JPO4. To assess the effects of PD, the tubules were treated with either amiloride (n = 6) or ouabain (n = 7). JPO4 fell significant in both circumstances with reduction of PD, although there remained significant residual transport following amiloride addition. The data demonstrate significant phosphate transport in this segment that is independent of PTH and altered by transepithelial electrical and concentration gradients as well as flow rate.