PMID- 1370611 OWN - NLM STAT- MEDLINE DCOM- 19920218 LR - 20190612 IS - 0006-291X (Print) IS - 0006-291X (Linking) VI - 182 IP - 1 DP - 1992 Jan 15 TI - Detection of thyrotropin-releasing hormone (TRH) mRNA by the reverse transcription-polymerase chain reaction in the human normal and tumoral anterior pituitary. PG - 182-7 AB - To investigate the presence of TRH mRNA in the human anterior pituitary tissue, total RNA from human normal and tumoral anterior pituitary, hypothalamus (positive control) and muscle tissues (negative control) was reverse transcribed (RT) to the first strand of cDNA. RT products were then amplified by polymerase chain reaction (PCR) using a set of three exon-specific primers (two external 5' and 3' primers and one internal 3' primer) for a target sequence of the TRH gene including an intronic sequence of about 650 base pairs (bp). Southern analysis of the RT-PCR products specifically hybridizing with a 45-mer TRH probe showed two bands of the predicted sizes (399 and 351 bp) far more intense in hypothalamus than in normal and tumoral anterior pituitary tissue. The 399 and 351 bp RT-PCR products contained the BglII enzyme restriction site included in the TRH cDNA sequences spanned by the primers and the two respective digested fragments which were, as predicted, 337 and 289 bp long, hybridized with the TRH probe. Based on these results, we can conclude that the RT-PCR products generated from RNA tissue were the target TRH sequences in the human normal and tumoral anterior pituitary tissue as well as in the hypothalamus. Our data imply TRH gene expression in the human anterior pituitary. FAU - Pagesy, P AU - Pagesy P AD - INSERM U. 223, Faculte de Medecine Pitie-Salpetriere, Paris, France. FAU - Croissandeau, G AU - Croissandeau G FAU - Le Dafniet, M AU - Le Dafniet M FAU - Peillon, F AU - Peillon F FAU - Li, J Y AU - Li JY LA - eng PT - Journal Article PL - United States TA - Biochem Biophys Res Commun JT - Biochemical and biophysical research communications JID - 0372516 RN - 0 (Oligodeoxyribonucleotides) RN - 0 (RNA, Messenger) RN - 5Y5F15120W (Thyrotropin-Releasing Hormone) RN - 9002-72-6 (Growth Hormone) RN - EC 2.7.7.49 (RNA-Directed DNA Polymerase) SB - IM MH - Adenoma/genetics/metabolism MH - Base Sequence MH - Blotting, Southern MH - Growth Hormone/metabolism MH - Humans MH - Hypothalamus/physiology MH - Molecular Sequence Data MH - Muscles/physiology MH - Oligodeoxyribonucleotides MH - Organ Specificity MH - Pituitary Gland, Anterior/*physiology/physiopathology MH - Pituitary Neoplasms/*genetics/metabolism MH - Polymerase Chain Reaction/methods MH - RNA, Messenger/analysis/*genetics MH - RNA-Directed DNA Polymerase MH - Reference Values MH - Restriction Mapping MH - Thyrotropin-Releasing Hormone/*genetics EDAT- 1992/01/15 00:00 MHDA- 1992/01/15 00:01 CRDT- 1992/01/15 00:00 PHST- 1992/01/15 00:00 [pubmed] PHST- 1992/01/15 00:01 [medline] PHST- 1992/01/15 00:00 [entrez] AID - S0006-291X(05)80128-7 [pii] AID - 10.1016/s0006-291x(05)80128-7 [doi] PST - ppublish SO - Biochem Biophys Res Commun. 1992 Jan 15;182(1):182-7. doi: 10.1016/s0006-291x(05)80128-7.