Structural, functional and pormorphic analysis of the regulatory regions necessary for the expression of the human neuronal tryptophan hydroxylase gene.
| Project/Area Number |
16591170
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Psychiatric science
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| Research Institution | St. Marianna University School of Medicine |
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Principal Investigator |
MATSUI Hiroaki St. Marianna University School of Medicine, Department of Medicine, Professor, 医学部, 教授 (90181685)
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| Co-Investigator(Kenkyū-buntansha) |
OSADA Kenichi St. Marianna University School of Medicine, Department of Medicine, Lecturer, 医学部, 講師 (20233504)
HIROI Tomoko St. Marianna University School of Medicine, Department of Medicine, Lecturer, 医学部, 講師 (20238398)
ASAKURA Mikio St. Marianna University School of Medicine, Department of Medicine, Associate Professor, 医学部, 助教授 (70103504)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2006: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Tryptophan hydroxylase 2 / Tryptophan hydroxylase 1 / Transcriptional regulation region / Rat serotonergic cell line RN46A / Neuron-restrictive silencer factor / Neuron-restrictive silencer element / Histone deacetylase / ヒト肺小細胞癌HSP-77細胞 / ヒト神経芽細胞腫SH-SY5Y細胞 / セロトニン神経特異的転写因子 / ヒト脳型トリプトファン水酸化酵素 / ヒト抹消型トリプトファン水酸化酵素 / ラット縫線由来RN46A細胞 / セロトニン系神経細胞特異的転写因子 |
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| Research Abstract |
Promoter polymorphism of the human tryptophan hydroxylase-2 (hTPH2) gene coding for the rate-limiting enzyme of serotonin (5-HT) synthesis in the brain is thought to be related to neuropsychiatric diseases as well as personality traits. Elucidation of mechanisms involved in the transcriptional regulation of the hTPH2 gene will assist our understanding of how the expression of the hTPH2 gene could be regulated in a tissue- and cell-type specific manner, and be altered by various pathologic and pharmacologic states. To address these questions more directly, we tested the hTPH2 gene promoter activity and found a critical element necessary for the hTPH2 gene expression. A 8.7-kb fragment upstream to the transcription start site was cloned into pGL4-Basic and a series of 5'-deletion mutants were constructed. Promoter activities were assessed by transient transfections into immortalized rat serotonergic RN46A cells. RT-PCR analysis confirmed the expression of serotonergic marker genes, trans
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cription factors Mash1, Nkx2.2, Lmx1b, GATA2, GATA3, Pet-1, and functional proteins 5-HTT and 5-HT1A receptor in RN46A cells. Unexpectedly, RN46A cells express TPH1 but not TPH2. Accordingly, hTPH2 gene promoter activities could not be measured. By close inspection of the promoter sequence, a repressor element 1 or neuron- restrictive silencer element (RE1/NRSE)-like sequence was found at the vicinity of the transcription start site. Gel mobility shift assay showed a RE1-silencing transcription factor or neuron-restrictive silencer factor (NRSF) may bind to this element. Either by introducing mutations into this element or treating RN46A cells with histone deacetylase inhibitors, hTPH2 gene promoter activities were able to be measured. These results suggest that the expression of the TPH2 gene is strictly controlled through the repressor-mediated regulatory mechanisms. The neuronal activity-dependent modulation of the repressor system may contribute to the transcriptional regulation of the TPH2 gene in the brain. Less
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Report
(4 results)
Research Products
(53 results)
