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 language | English |
|---|---|
| Pages (from-to) | 230-235 |
| Number of pages | 6 |
| Journal | Journal of Endocrinological Investigation |
| Volume | 19 |
| Issue number | 4 |
| Publication status | Published - 1996 |
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Keywords
- Thyroid cens transfection
- Tsh receptor
ASJC Scopus subject areas
- Endocrinology
Cite this
Transfection with the cDNA of the human thyrotropin receptor of a poorly differentiated rat thyroid cell line (FRT). / Elisei, R.; Pinchera, A.; Chiovato, L.; Mammoli, C.; Agretti, P.; Romei, C.; Santini, F.; Bendinelli, G.; Fiore, E.; Capaccioli, A.; Vitti, P.
In: Journal of Endocrinological Investigation, Vol. 19, No. 4, 1996, p. 230-235.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Transfection with the cDNA of the human thyrotropin receptor of a poorly differentiated rat thyroid cell line (FRT)
AU - Elisei, R.
AU - Pinchera, A.
AU - Chiovato, L.
AU - Mammoli, C.
AU - Agretti, P.
AU - Romei, C.
AU - Santini, F.
AU - Bendinelli, G.
AU - Fiore, E.
AU - Capaccioli, A.
AU - Vitti, P.
PY - 1996
Y1 - 1996
N2 - 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.
AB - 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.
KW - Thyroid cens transfection
KW - Tsh receptor
UR - http://www.scopus.com/inward/record.url?scp=0030115304&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030115304&partnerID=8YFLogxK
M3 - Article
C2 - 8862503
AN - SCOPUS:0030115304
VL - 19
SP - 230
EP - 235
JO - Journal of Endocrinological Investigation
JF - Journal of Endocrinological Investigation
SN - 0391-4097
IS - 4
ER -