Micropuncture studies demonstrate phosphate reabsorption in proximal tubules and between the late proximal and early distal convoluted tubule accessible to micropuncture. To further define the sites of phosphate reabsorption, the stationary microperfusion technique was applied to proximal and distal nephron segments. Phosphate reabsorption was evaluated in superficial loops of proximal tubules, descending segments beyound late proximal tubules accessible to micropuncture, ascending segments up to the point of micropuncture in the distal tubule, and superficial loops of distal tubules of thyroparathyroidectomized rats. Microperfusates of 1.3 or 2.6 nl (100 mmol/1 mannitol, 100 mmol/l NaCL, 32P-phosphate and 3H-inulin) were injected and then withdrawn after contact times of 2--108 s. Phosphate recovery relative to that of inulin was determined. A steep exponential decline of phosphate recovery (R) iwth increasing contact time (t) was observed in the superficial proximal tubule and descending segments. The slopes of the logarithmic regressions (10log R)/t, +/- SEM) were: -1.68 +/- 0.33 and -1.21 +/- 0.24min-1 in superficial proximal tubules and descending segments respectively. In contrast, no significant decline in phosphate recoveries (-0.02 +/- 0.04 and + 0.11 +/- 0.10 min-1) was apparent in the ascending segments and distal tubule. It is concluded that phosphate is reabsorbed in the proximal convoluted tubule and adjacent descending segments of the superficial nephron and that there is no significant phosphate reabsorption in distal convoluted tubules and adjacent ascending segments.