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Thyrotropin receptor (TSH-R) has been thought to be
thyroid-specific, but, by Northern blot analysis, we found that rat
adipose tissue expressed TSH-R mRNAs in amounts approaching those in
the thyroid. To investigate the function of TSH-R from adipose tissue,
we screened a rat fat cell
Volume 270,
Number 18,
Issue of May 5, pp. 10833-10837, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
gt11 cDNA library for TSH-R sequences
using a
P-labeled rat thyroid TSH-R cDNA as a probe. Among
10
plaques, we obtained four positive clones. Sequencing of
these cDNAs has revealed that two of them (F
and F
) contained
both initiation and termination codons. Comparison of F with the
thyroid TSH-R cDNA sequence revealed that F was almost identical
to the thyroid TSH-R, except that nucleotides 1041 and 1277 were
changed from A to G and from C to T, respectively. In contrast, we
found that F contained 21 novel nucleotides between nucleotides
467 and 468 of the thyroid TSH-R cDNA, encoding an additional 7 amino
acids. However, when we prepared mRNA from adipose tissue and
transcribed it into cDNA, we failed to amplify the F type of TSH-R
cDNA by polymerase chain reaction, suggesting that F mRNAs are
rare in the tissue. We then ligated F cDNAs into pSG5 and transfected
them with pSV
-neo into Chinese hamster ovary (CHO)-K1
cells. TSH stimulated cAMP formation in CHO-F cells in a manner
similar to that in CHO cells transfected with thyroid TSH-R cDNA. In
contrast, no increase of cAMP was observed in CHO-F cells. IgG
from patients with Graves' disease ( n = 4) showed
thyroid-stimulating antibody activity only in CHO-F cells
(1288-4582%). In addition, CHO-F cells and CHO cells
transfected with thyroid TSH-R showed similar I-TSH
binding activity. These results indicate that the fat cell expresses
high levels of a TSH-R whose function is indistinguishable from that in
the thyroid and suggest that the TSH-R autoantibody plays an important
role in the pathogenesis of the extrathyroidal manifestations of
Graves' disease.
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