Studies were performed in the rat to determine the transepithelial potential difference (PD) of the medullary collecting duct (MCD). With the microcatheter as an exploring electrode the PD from duct lumen to the reference electrode, placed in peritoneal fluid, was approximatley 1 mV, lumen positive, in the MCD. It was unlikely that the polarity and small magnitude of PD was due to an artifact created by the microcatheter, since essentially similar results were obtained by transmural puncture of the inner third of the duct using Ling-Gerard glass electrodes and glass microelectrodes, tip OD 3-5 micron. Moreover, a PD of a few millivolts, lumen positive, was observed in experimental states characterized by marked increases in potassium and hydrogen ion secretion and sodium absorption. Further studies showed that the diffusion potential existing within the interstitial fluid compartment, between papilla and reference electrode, was approximately +2 mV, owing to the higher NaCl concentration in the papillary area. These data demonstrate, therefore, that the transepithelial PD in MCD approaches zero in control animals, and is not substantially altered by marked changes in net electrolyte transport.