RENAL UPTAKE AND DEGRADATION OF TRAPPED-LABEL CALCITONIN

In order to quantitate the role of the kidneys in the clearance and degradation of calcitonin, a trapped-label procedure was used to label human calcitonin. In contrast to conventional [I-125]calcitonin, the trapped-label preparation allows quantitative measurements of the extent of uptake as well as of degradation in vivo because the final degradation products do not leave the cells. Trapped-label calcitonin activated adenylate cyclase of bone cells and kidney, as did the native hormone. Ten minutes after intravenous injection into rats, 16% of a trace dose was found in the kidneys. Renal recovery increased to 20% after one hour; in addition, 14% of the injected dose was found in the urine. Eighty per cent of the radioactivity in the urine was in high-molecular weight material. After 90 min, the sum of the accumulated radioactivities in the kidneys and the urine reached 40% of the dose. More than 80% of the radioactivity was sedimentable by centrifuging in a density gradient, indicating that intact calcitonin, as well as the degradation products in the cells, were enclosed within membrane-bound vesicles. Two minutes after injection of trapped-label calcitonin, the peak of radioactivity was found in light gradient fractions associated with cell membrane marker enzymes. Between 5 and 15 min, the peak migrated from light fractions to heavy fractions containing lysosomal marker enzymes. After just 2.5 min, 61% of the renal radioactivity was in low-molecular weight degradation products, as determined by gel filtration. The kinetics of renal degradation of calcitonin indicate that substantial amounts of endocytosed calcitonin is degraded before the hormone reaches the lysosomes.