Functional significance of morphological heterogeneities along the thick ascending limb of Henle's loop of hamsters was explored by the in vitro microperfusion technique with special reference to K+ transport. The transmission electron microscopic study confirmed that there are two types of cells, with smooth surface (S-cell) and rough surface (R-cell), respectively, and that the former is abundant in the medullary thick ascending limb (MTAL), whereas the latter is in the cortical portion (CTAL). The electrophysiological study revealed that in both segments there are two cell populations, one having high basolateral and low apical membrane K+ conductances (HBC) and the other having low basolateral and high apical K+ conductances (LBC). Random cell puncture revealed that the ratios of HBC/LBC were 24/7 (77%/23%) in the MTAL and 7/22 (24%/76%) in the CTAL, suggesting that HBC corresponds to S-cell, whereas LBC corresponds to R-cell. Net K+ transport was determined in two segments by measuring K+ concentration in the collected and perfused fluid by ultramicroflame photometry. In all six tubules of MTAL, net K+ flux had a direction to reabsorption with a mean of 4.87 +/- 0.46 pmol.min-1.mm-1. In marked contrast, in all six tubules of CTAL, we observed K+ secretion with a mean of -3.81 +/- 0.49 pmol.min-1.mm-1. The transmural voltage was positive in both segments and was significantly higher in the CTAL (7.8 +/- 0.5 mV) than in the MTAL (2.5 +/- 0.2 mV). From these observations, we conclude that the S-cell corresponding to the HBC cell reabsorbs K+, whereas the R-cell corresponding to the LBC cell secrets K+.(ABSTRACT TRUNCATED AT 250 WORDS)