| Project/Area Number |
14370025
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Environmental physiology (including Physical medicine and Nutritional physiology)
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| Research Institution | Nippon Medical School |
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Principal Investigator |
SAKUMA Yasuo Nippon Medical School, Grad Sch Med, Professor, 大学院・医学研究科, 教授 (70094307)
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| Co-Investigator(Kenkyū-buntansha) |
KATO Masakatsu Nippon Medical School, Faculty of Medicine, Associate Professor, 医学部, 助教授 (90143239)
KIYAMA Yuko Nippon Medical School, Faculty of Medicine, Assistand Professor, 医学部, 講師 (60234390)
ORIKASA Chitose Nippon Medical School, Faculty of Medicine, Research Associate, 医学部, 助手 (20270671)
HAMADA Tomohiro Nippon Medical School, Faculty of Medicine, Research Associate, 医学部, 助手 (90312058)
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| Project Period (FY) |
2002 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥14,600,000 (Direct Cost: ¥14,600,000)
Fiscal Year 2005: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2003: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2002: ¥5,600,000 (Direct Cost: ¥5,600,000)
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| Keywords | estrogen receptor alpha / promoter transgenic rat / sexually dimorphic nucleus / EGFP / medial preoptic area / stria terminalis / immunohistochemistry / membrane receptor / エストロゲン受容体 / プロモーター / トランスジェニック / トランスジェニックラット / 可視化 / プロモータ / 視床下部 / 海馬 |
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| Research Abstract |
Transgenic rats expressing enhanced green fluorescent protein (EGFP) under the control of an estrogen receptor (ER) alpha promoter were generated to tag ERalpha-positive neurons in the brain. Two transgenes, one containing sequences for promoter A and DsRed and the other containing sequences for promoter 0/B and EGFP, were injected simultaneously into Wistar rat zygotes. Twenty-two founders with both transgenes were identified. Ten lines of these founders expressed the EGFP tag in the brains of their first filial generation, whereas none similarly expressed the DsRed tag. In two lines selected for the brightness of the EGFP fluorescence in their brains, tagged cells showed essentially the same patterns. Tagged cells were in the preoptic area (POA), bed nucleus of the stria terminalis (BNST), hypothalamic arcuate nucleus and medial amygdala. ERalpha-immunoreactive neurons were identified in all of these structures by immunohistochemistry. In ovariectomized females, approximately 75% of the EGFP-fluorescent cells in the POA-BNST were immunoreactive for ERalpha. In the POA-BNST, ovariectomy increased the number of EGEP-immunopositive cells and estrogen supplementation reversed this effect, indicating that the promoter 0/B is involved in estrogen-induced downregulation of ERalpha. EGFP was also present in cells in the cerebral cortex and hippocampus, which have not previously been associated with endocrine regulation. Conversely, only a few cells were tagged in the hypothalamic ventromedial nucleus, which contained many ERalpha-immunoreactive neurons. This discrepancy could have arisen as a result of differential promoter usage.
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