Collection and analysis of absorbate from proximal straight tubules

Abstract

When isolated segments of rabbit proximal straight tubules were perfused under oil at 37 degrees C, we observed that droplets of absorbate formed on the peritubular surface. Volume absorption under these conditions was the same as with customary aqueous bathing solutions when calculated either from the rate of absorbate formation (0.39 +/- 0.03 nl X min-1 X mm-1) or from the difference in measured perfusion and collection rates (0.37 +/- 0.04 nl X min-1 X mm-1). Absorbate formation continued at a steady rate for at least 2 h but was inhibited by 71% at 28 degrees C. The absorbate was found to have a composition that differed from the perfusate, as would be expected in the presence of preferential absorption of glucose, amino acids, and HCO-3. The Cl- concentration in the absorbate was 11.2 +/- 1.8 mM less than in the perfusate. The glucose concentration in the absorbate was 4.5 mM compared with 0.9 mM in the perfusate. Finally, the nonmetabolizable amino acid cycloleucine, which was added to the perfusate at 0.35 mM, had a concentration of 2.9 mM in the epithelial cells compared with 1.6 mM in the absorbate. These data establish the usefulness of this technique for examining solute and water absorption in the proximal nephron and show that the absorbate can differ considerably in solute composition from the luminal perfusate.