The functional expression of recombinant human thyrotropin receptors in nonthyroidal eukaryotic cells provides evidence that homologous desensitization to thyrotropin stimulation requires a cell-specific factor.
TSH desensitization involves decreased coupling of the TSH receptor to the adenylate cyclase regulatory protein, Gs. There is evidence that a desensitization protein in thyroid cells plays a role in this process. The molecular cloning of the human TSH receptor and its stable expression in Chinese hamster ovary (CHO-TSHR) cells allowed us to test whether or not TSH desensitization can occur in a nonthyroidal cell. Similar to human thyroid cells, maximal stimulation of cAMP levels in CHO-TSHR cells was attained after 30-60 min of exposure to bovine TSH. Unlike in human thyroid cells, however, preincubation of CHO-TSHR cells with TSH for 12-16 h did not decrease the subsequent cAMP response to a 1-h pulse of TSH stimulation. That is, the human TSH receptor in CHO-TSHR cells does not undergo functional desensitization. Scatchard plot analysis of specific TSH binding to the CHO-TSHR cells revealed high and low affinity sites (Ka of 1.8 +/- 0.4 x 10(9) M-1 and 1.4 +/- 0.3 x 10(7) M-1, respectively), with approximately 10(5) TSH receptors per cell. This is 10- to 100-fold greater than the number of TSH receptors estimated to be present on human thyroid cells. Untransfected CHO cells exhibited only the low affinity binding site. Prior exposure of CHO-TSHR cells to bovine TSH or to (Bu)2cAMP for periods up to 24 h did not reduce [125I]TSH binding to these cells. In summary, desensitization of the adenylate cyclase response to TSH stimulation does not occur in nonthyroidal cells expressing a human TSH receptor with normal functional and TSH binding characteristics. These data support the concept that a cell-specific protein may be involved in homologous TSH desensitization.
Chazenbalk
G D
GD
Thyroid Molecular Biology Unit, Veterans Administration Medical Center, San Francisco, California 94121.
Nagayama
Y
Y
Kaufman
K D
KD
Rapoport
B
B
D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D000242:Cyclic AMP / Q000096:biosynthesis; D004361:Drug Tolerance; D019204:GTP-Binding Proteins / Q000502:physiology; D015870:Gene Expression; D006801:Humans; D011989:Receptors, Thyrotropin / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics*; D013972:Thyrotropin / Q000378:metabolism / Q000494:pharmacology*; D014162:Transfection
D011989:Receptors, Thyrotropin; D011994:Recombinant Proteins; D013972:Thyrotropin; D000242:Cyclic AMP; D019204:GTP-Binding Proteins