Structure of the thyrotrophin-releasing hormone receptor in human pituitary adenomas.
E Faccenda, S Melmed, JS Bevan… - Clinical …, 1996 - europepmc.org
E Faccenda, S Melmed, JS Bevan, KA Eidne
Clinical endocrinology, 1996•europepmc.orgMethods Genomic DNA was extracted from adenoma and lymphocyte samples and the
entire coding region of the TRH receptor was amplified using 5 overlapping pairs of PCR
primers. The PCR products were analysed for mutations by non-denaturing polyacrylamide
gel electrophoresis which reveals single-strand conformational polymorphisms (SSCP) as a
mobility shift in product migration. Wild-type and mutant TRH receptor cDNA were similarly
analysed to confirm the sensitivity of the method. Additionally, PCR products were ligated …
entire coding region of the TRH receptor was amplified using 5 overlapping pairs of PCR
primers. The PCR products were analysed for mutations by non-denaturing polyacrylamide
gel electrophoresis which reveals single-strand conformational polymorphisms (SSCP) as a
mobility shift in product migration. Wild-type and mutant TRH receptor cDNA were similarly
analysed to confirm the sensitivity of the method. Additionally, PCR products were ligated …
Methods
Genomic DNA was extracted from adenoma and lymphocyte samples and the entire coding region of the TRH receptor was amplified using 5 overlapping pairs of PCR primers. The PCR products were analysed for mutations by non-denaturing polyacrylamide gel electrophoresis which reveals single-strand conformational polymorphisms (SSCP) as a mobility shift in product migration. Wild-type and mutant TRH receptor cDNA were similarly analysed to confirm the sensitivity of the method. Additionally, PCR products were ligated into a PCR cloning vector and DNA sequencing carried out to confirm the findings of SSCP analysis.
Results
The human TRH receptor retained normal wild-type sequence in the large group of TSH secreting, PRL secreting, GH secreting and non-functioning pituitary adenomas investigated in this study.
Conclusion
Our observations suggest that the TRH receptor structure is normal in TSH secreting, PRL secreting, GH secreting and non-functioning pituitary adenomas. It is therefore unlikely that the TRH receptor is involved in the pathology associated with the types of pituitary adenomas investigated in this study. It is possible that some other component of the pathway controlling TRH-signalling events may be implicated in pituitary tumorigenesis.
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