MODULATION OF CELL NUCLEOTIDE LEVELS OF ISOLATED KIDNEY-TUBULES

The mechanisms responsible for the large increases of intracellular ATP levels seen after isolated rabbit proximal tubules are treated with exogenous adenine nucleotides were studied. Exogenous ATP was rapidly degraded via adenosine as far as hypoxanthine. Degradation of AMP to adenosine was substantially inhibited by .beta.-glycerol purines to increase intracellular ATP levels, single large doses of adenosine were less effective than equimolar doses ofa exogenous ATP but were substantially more effective than exogenous inosine or hypoxanthine. Exogenous guanine derived compounds increased ony cell GTP. Incremental delivery of smaller doses of adenosine to maintain medium levels greater than 5 .mu.M or inhibition of adenosine deaminase with erythro-9-[3-(2-hydroxynonyl)]adenine or 2''-deoxycoformicin enhanced the nucleoside''s effectiveness. However, the initial increase of cell ATP was still greater after treatment with exogenous ATP than after adenosine and, in the presence of adenosine deaminase inhibition, larger increases of cell ATP were produced by 50 .mu.M adenosine than by 250 .mu.M adenosine. These observations are most consistent with substrate inhibition of adenosine kinase by adenosine. Furthermore, the adenosine kinase inhibitor, 5-iodotubercidin, prevented the increases of cell ATP resulting from exogenous adenosine or exogenous ATP. These studies demonstrate how the differential uptake and utilization characteristics of nucleosides and bases can fully account for the increases of intracellular nucleotides produced in isolated tubules by exogenous purines.