Transfection with the cDNA of the human thyrotropin receptor of a poorly differentiated rat thyroid cell line (FRT)

R. Elisei, A. Pinchera, L. Chiovato, C. Mammoli, P. Agretti, C. Romei, F. Santini, G. Bendinelli, E. Fiore, A. Capaccioli, P. Vitti

Research output: Contribution to journalArticlepeer-review

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 corre-spending 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 3Hthymidine incorporation into DMA. 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.

Original languageEnglish
Pages (from-to)230-235
Number of pages6
JournalJournal of Endocrinological Investigation
Volume19
Issue number4
Publication statusPublished - 1996

Keywords

  • Thyroid cens transfection
  • Tsh receptor

ASJC Scopus subject areas

  • Endocrinology

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