title: A novel TRH-PFTAIRE protein kinase 1 pathway in the cerebellum: subtractive hybridization analysis of TRH-deficient mice.
abstract: TRH has been reported to possess several neurophysiological actions in the brain. To gain insights into the molecular mechanisms underlying these effects, particularly in the cerebellum, we attempted to clone a cDNA that was regulated by TRH using TRH knockout mice and subtractive cDNA analysis. Over 100 clones obtained by subtractive hybridization analysis between the wild-type and TRH-1-cerebellum were analyzed. Four clones among them were identical and cdc2-related kinase (PFTAIRE protein kinase 1 (PFTK1)) cDNA, which was previously reported to be expressed only in the brain and testis. PFTK1 mRNA levels in the euthyroid TRH-1- cerebellum supplemented with thyroid hormone were significantly decreased compared with those in the wild-type. Induction of PFTK1 mRNA by TRH was also observed in a time- and dose-dependent manner in human medulloblastoma-derived HTB-185 cells that expressed TRH receptor subtype I mRNA. In addition, treatment of 8-Br-cGMP significantly increased PFTK1 mRNA levels, and a specific inhibitor of cGMP production, ODQ, completely blocked TRH-induced expression of PFTK1 mRNA. Furthermore, induction of PFrK1 mRNA by TRH was significantly inhibited by a NOS specific inhibitor, L-NAME, but not by a MEK inhibitor, PD98059 or a calcium channel inhibitor, nimodipine. These findings demonstrated, for the first time, a novel pathway between a neuropeptide and a cell cycle related peptide in the brain, and PFTK1 may be a key regulator for TRH action in t he cerebellum through t he NO-cGMP pathway.
authors.affiliation: First Department of Internal Medicine, Gunma University School of Medicine, Maebashi, Japan.
authors.forename: Tetsu
authors.initials: T
authors.lastname: Hashida
authors.forename: Masanobu
authors.initials: M
authors.lastname: Yamada
authors.forename: Koshi
authors.initials: K
authors.lastname: Hashimoto
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Shibusawa
authors.forename: Tsuyoushi
authors.initials: T
authors.lastname: Monden
authors.forename: Teturou
authors.initials: T
authors.lastname: Satoh
authors.forename: Masatomo
authors.initials: M
authors.lastname: Mori
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000502:physiology; D015152:Blotting, Northern; D002478:Cells, Cultured; D002531:Cerebellum / Q000502:physiology*; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000096:biosynthesis / Q000494:pharmacology; D018844:Cyclin-Dependent Kinases; D029721:Drosophila Proteins; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D011494:Protein Kinases / Q000235:genetics* / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013973:Thyrotropin-Releasing Hormone / Q000172:deficiency / Q000235:genetics* / Q000502:physiology*; D014157:Transcription Factors; D014407:Tumor Cells, Cultured
chemical_list: D029721:Drosophila Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D013973:Thyrotropin-Releasing Hormone; D000242:Cyclic AMP; D011494:Protein Kinases; D018844:Cyclin-Dependent Kinases; C408322:Eip63E protein, Drosophila