Abstract
A cell line derived from the Fisher rat thyroid (FRT), that does not have functional TSH receptor, was stably transfected with the cDNA of the human TSH receptor (hTSH-R). In wild FRT cells TSH (1–1000 mU/l) was unable to increase cAMP production, while 10–10000 nmol/l forskolin elicited a 10–30 fold cAMP stimulation. Two of the transfected clones were responsive to TSH in terms of cAMP production. In particular, the FRT-R3 transfected clone showed the highest sensitivity to the hormone with a 10 fold cAMP increase over the basal at 100 mU/l TSH. The Northern blot analysis using a 2.4 kbp cDNA probe for the hTSH-R showed a band corresponding to the mRNA of TSH receptor in FRT-R3 cells, but not in wild FRT cells. In both cell types TSH was ineffective in stimulating growth assayed by 3H-thymidine incorporation into DNA. Hybridization with a probe for thyroperoxidase on polymerase chain reaction products after reverse transcription of mRNA showed that FRT-R3, as well as FRT cells, do not have a transcript for thyroperoxidase. In conclusion, the data reported in this paper show that the insertion of the hTSH-R cDNA in the genome of poorly differentiated rat thyroid cells results in the recovery of TSH-dependent adenylate cyclase, but not other differentiated thyroid cell functions.
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Elisei, R., Pinchera, A., Chiovato, L. et al. Transfection with the cDNA of the human thyrotropin receptor of a poorly differentiated rat thyroid cell line (FRT). J Endocrinol Invest 19, 230–235 (1996). https://doi.org/10.1007/BF03349873
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DOI: https://doi.org/10.1007/BF03349873