Handling of lysozyme in isolated perfused proximal tubules

Abstract

Isolated, perfused proximal tubules from rabbit were used to study the luminal endocytic uptake, digestion, and transcellular transport of 125I-lysozyme. Ten tubules were perfused for 20 min with 125I-lysozyme and [14C]inulin and then with tracer-free perfusate for additional 40 min before fixation. The uptake and digestion of lysozyme was calculated per millimeter tubule length. The transfer of intact lysozyme from perfusate to the bath was measured and compared with the transfer of inulin. Five tubules were processed for electron microscope autoradiography, and the grain distribution was analyzed quantitatively. The results show that 2.7% of the perfused amount of lysozyme was taken up, and 21.3% of the absorbed protein was digested. The present experiments demonstrate that the transfer of intact lysozyme from lumen to bath is not significantly different from the transfer of inulin. The autoradiographic analysis showed that lysozyme was localized mainly in endocytic vacuoles and lysosomes after 60 min of perfusion.