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A Novel Transcript for the Thyrotropin-Releasing Hormone Receptor in Human Pituitary and Pituitary Tumors

First Department of Internal Medicine (M.Y., K.H., T.S., N.S, M.M.) and Department of Neurosurgery (H.K.), Gunma University School of Medicine, Maebashi, Gunma; and the Departments of Endocrinology and Metabolism (Y.O.) and Neurosurgery (S.Y.), Toranomon Hospital, Tokyo, Japan

Address all correspondence and requests for reprints to: Masanobu Yamada, M.D., Ph.D., First Department of Internal Medicine, Gunma University School of Medicine, 3–39-15 Showa-machi, Maebashi, Gunma 371, Japan. E-mail: myamada{at}sb.gunma-u.ac.jp

We measured the amounts of TRH receptor (TRHR) messenger ribonucleic acid (mRNA) in human normal pituitary and pituitary tumors and found a novel transcript of the TRHR gene. Competitive PCR revealed expression of the TRHR mRNA in all pituitary adenomas examined, and its level was variable and similar to that in the normal pituitary. When the C-terminal region was amplified by PCR, an additional short product was observed. Cloning and sequence analysis of this short fragment revealed that the deleted sequence corresponded exactly to the 5'-sequence of exon 3, indicating alternative splicing of the TRHR mRNA. This alternative splicing resulted in a frame shift, yielding a C-terminal truncated protein (HTRHR2) on translation. Expression analysis of HTRHR2 in Chinese hamster ovary cells showed no significant binding to [³H]MeTRH or response of intracellular calcium to TRH administration. However, the mRNA ratio of HTRHR2 vs. the wild type (HTRHR1) was significantly different among pituitary tumors. The highest ratio was observed in prolactinomas (30%), and almost no detectable expression was found in GH-producing tumors. These findings indicate that this novel transcript of the human TRH receptor gene is produced in a tumor-specific manner and may be a useful parameter for evaluation of individual pituitary tumors.

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