1999 Fiscal Year Final Research Report Summary
Real-time imaging of steroid hormone receptor dynamics in living cells with GFP
| Project/Area Number |
10680748
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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 |
Neuroscience in general
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
NISHI Mayumi Kyoto Prefectural Univ of Med, Dept of Medicine, Assistant professor, 医学部, 講師 (40295639)
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| Co-Investigator(Kenkyū-buntansha) |
MORITA Noriyuki Kyoto Prefectural Univ of Med, Dept of Medicine, Assistant professor, 医学部, 講師 (50239662)
OZAWA Hitoshi Kyoto Prefectural Univ of Med, Dept of Medicine, Assistant professor, 医学部, 助教授 (60169290)
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| Project Period (FY) |
1998 – 1999
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| Keywords | steroid hormone receptor / transcription factor / primary neural culture / GFP / fusion protein / real time imaging / CCD camera / nuclear translocation |
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| Research Abstract |
Corticosteroids exert markedly diverse effects on development, differentiation, aging and regeneration in the nervous system. These effects are mediated via two receptor system in the brain, glucocorticoid receptor (GR) and mineralocorticoid receptor (MR). The expression of GR occurs in most neurons and glial cells throughout the brain, while MRs are localized mainly in the hippocampus. Although it is mostly accepted that GR is translocated into the nucleus from the cytoplasm only after ligand binding, the subcellular localization of MR is still quite controversial. To elucidate the intracellular trafficking of GR and MR in response to various conditioned in living cells, a chimera construct of a green fluorescent protein (GFP) and rat GR (GFP-GR) and MR (GFP-MR) were prepared. We transiently transfected the chimera construct into primary cultured rat hippocampal neurons, cortical glial cells, and non-neural cells such as COS-1 cells, and examined dynamic changes in subcellular localiz
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ation of GFP-GR and GFP-MR in these cells. GFP-GR was predominantly localized in the cytoplasm in the absence of ligand, while GRP-MR was distributed in both cytoplasm and nucleus in the majority of the transfected cells without ligand. Corticosterone (CORT), the principle corticosteroid of the rat, induced a rapid nuclear accumulation of GFP-GR and GFP-MR in all cell types used. Furthermore, we investigated the differential effects of CORT on the nuclear translocation pattern of corticosteroid receptors in single living cells co-transfected with cyan fluorescent protein (CFP)-GR and yellow fluorescent protein (YFP)-MR.
Our findings clearly showed that the subcellular distributions of GFP-GR and GFP-MR are dynamically changed in response to extracellular and intracellular conditions, and that there are no conspicuous variations in the manner of trafficking of GFP-GR and GFP-MR between neural cells and non-neural cells. We also demonstrated that CORT-induced nuclear accumulation of CFP-GR and YFP-MR exhibited differential time-course under different ligand concentrations and cell types. Less
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Research Products
(14 results)
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[Publications] Kawata,M., Yuri,K., Nishi,M., Ito,T., Hu,Z., Lu,H. and Yoshida,M.: "Steroid hormones and their receptors in the brain."J. Steroid Biochem. Molec. Biol.. 65. 273-280 (1998)
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「研究成果報告書概要(欧文)」より
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[Publications] Ozawa,H., Lion,J., Xie,C-X, Nishi,M., Steinbusch,H. and Kawata,M.: "Down-regulation of ACTH and glucocorticoid receptor immunoreactivity in hypothalamic arcuate neurons after adrenalectomy in the rat."NeuroReport. 10. 1571-1575 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Kanemasa,H., Ozawa,H., Konishi,H., Ito,T., Nishi,M., Mitsufuji,S., Kodama,T., Hattori,T. and Kawata,M.: "Distribution of glucocorticoid receptor immunoeactivity in the gastric mucosa of normal and adrenalectomized rats."Digestive Disease and Science. 44. 2081-2087 (1999)
Description
「研究成果報告書概要(欧文)」より
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