Homologous desensitization of cultured chick kidney cells to parathyroid hormone.
The development of refractoriness of the cAMP response to PTH in primary cultures of chick kidney cells and recovery from the refractory state was investigated. When cells were preincubated with bovine PTH1-34, complete refractoriness to a subsequent challenge with the hormone developed within 2 h and at hormone concentrations as low as 5 ng/ml. The ability of PTH to stimulate activation of cAMP-dependent protein kinase was also abolished by preincubation with the hormone. When cells were desensitized and then incubated in hormone-free medium, recovery of the cAMP response began within an hour and was maximal, but not complete (80%) after 16 h. Cycloheximide did not affect either desensitization or the rate or extent of recovery from the refractory state. Low concentrations of forskolin (2.5 X 10(-7) M) greatly enhanced cAMP production stimulated by PTH and higher concentrations (10(-6) - 10(-4) M) stimulated rates of cAMP production 50 times those obtained with PTH alone. Preincubation with forskolin did not bring about desensitization to PTH nor did preincubation with PTH affect the subsequent response to forskolin. The half-life of biologically active bovine PTH1-34 in chick kidney cell culture was approximately 12 h and the rate of its removal was not significantly altered during a 20-h incubation period. The results suggest that desensitization of chick kidney cells to PTH is not suggest that desensitization of chick kidney cells to PTH is not brought about by cAMP generation itself, is not primarily dependent on protein synthesis, and does not involve a change in the rate of removal of biologically active hormone from the medium. In addition, recovery of the cAMP response to PTH also does not require new protein synthesis. These results are compatible with a mechanism of desensitization which occurs at the level of the receptor or hormone-receptor coupling to adenyl cyclase.
D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D005576:Colforsin; D000242:Cyclic AMP / Q000096:biosynthesis*; D004224:Diterpenes / Q000494:pharmacology; D004361:Drug Tolerance; D004789:Enzyme Activation / Q000187:drug effects; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism; D007700:Kinetics; D010281:Parathyroid Hormone / Q000494:pharmacology*; D010446:Peptide Fragments / Q000494:pharmacology*; D011494:Protein Kinases / Q000378:metabolism*
D004224:Diterpenes; D010281:Parathyroid Hormone; D010446:Peptide Fragments; C025076:parathyroid hormone (1-34), bovine; D005576:Colforsin; D000242:Cyclic AMP; D011494:Protein Kinases