| Project/Area Number |
16601002
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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 |
細胞死(アポトーシス)
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| Research Institution | Yamaguchi University |
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Principal Investigator |
MIZUKAMI Yoichi Yamaguchi Univ., Center for Gene Res, Associate Prof., 総合科学実験センター, 助教授 (80274158)
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| Co-Investigator(Kenkyū-buntansha) |
HATANO Naoya Kagawa Univ., Rare Suger Res.Center, Associate Prof., 希少糖研究センター, 客員助教授 (10332280)
AKI Toshihiko Yamaguchi Univ., Center for Gene Res, Res.Associate, 総合科学実験センター, 助手 (60304474)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Estrogen / GPR30 / membrane / がん / プロテオーム / リガンド探索 |
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| Research Abstract |
Recently, GPR30 was reported to be a novel estrogen receptor ; however, its intracellular localization has remained controversial. To investigate the intracellular localization of GPR30 in vivo, we produced four kinds of polyclonal antibodies for distinct epitopes on GPR30. Immunocytochemical observations using anti-GPR30 antibody and anti-FLAG antibody show that FLAG-GPR30 localizes to the plasma membrane 24 hr after transfection.
Treatment with estrogen (17β-estradiol or E2) causes an elevation in the intracellular Ca^<2+> concentration ([Ca^<2+>]_i) within 10 sec in HeLa cells expressing FLAG-GPR30. In addition, E2 induces the translocation of GPR30 from the plasma membrane to the cytoplasm by 1 hr after stimulation. Immunohistochemical analysis shows that GPR30 exists on the cell surface of CA2 pyramidal neuronal cells. The images on transmission electron microscopy show that GPR30 is localized to a particular region on the associated with the plasma membranes of the pyramidal cells. These data indicate that GPR30,a transmembrane receptor for estrogen, is localized to the plasma membrane of CA2 pyramidal neuronal cells of the hippocampus in rat brain.
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