Liquid hybridization analysis of TSH receptor mRNA in normal and abnormal human thyroid tissues

Radiolabeled human TSH receptor (hTSHR) cRNA probes encoding nucleotides 37-2298 and 37-209, with unlabeled sense RNA control segments, were used in a liquid hybridization assay and found to be highly specific and sensitive enabling detection of 0.5 fmol of hTSHR mRNA. Using normal human thyroid monolayer cell cultures we calculated that the average number of TSHR mRNA transcripts was 95 +/- 5 per cell under in vitro basal conditions. We found no significant difference between the hTSHR mRNA concentrations of intact normal human thyroid tissue (n = 4) and specimens from patients with multinodular goiter (n = 5) and Graves' disease thyroid tissues (n = 5) (23.0, 25.2, and 27.6 fmol of hTSHR and mRNA/mg total cellular RNA, respectively). However, there was a relative deficiency of hTSHR mRNA in some samples of thyroid papillary carcinoma tissue (n = 5) (12 fmol of hTSHR mRNA/mg total RNA, p < 0.05). The hTSHR 37-2298 probe was fully protected in normal and abnormal thyroid tissues, consistent with the absence of large deletions or insertions in the hTSHR mRNA transcripts but additional bands were present, consistent with the production of splicing variants.