| Project/Area Number |
15590206
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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 |
Environmental physiology (including Physical medicine and Nutritional physiology)
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| Research Institution | University of Yamanashi |
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Principal Investigator |
ARITA Jun University of Yamanashi, Department of Research Interdisciplinary Graduate School of Medicine & Engineering, professor, 大学院・医学工学総合研究部, 教授 (80128587)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIDA Maho University of Yamanashi, Department of Research Interdisciplinary Graduate School of Medicine & Engineering, research associate, 大学院・医学工学総合研究部, 助手 (80362086)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | estrogen / cell proliferation / prolactin / anterior pituitary gland / estrogen receptor |
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| Research Abstract |
Research project 1
1)Using rat anterior pituitary cells in culture, we established triple immunostaining for Ki67, estrogen receptor (ER), and prolactin (PRL).
2)When estrogen stimulated proliferation of lactotrophs, the proportion of ER-imnumoreadive cells in proliferating lactctrophs was decreased. The proportion of the ER-immunoreactive cells was not changed when estrogen inhibited IGF-1-induced proliferation.
Research project 2
1)We produced recombinant adenovirus vectors that carry PRL promoter-driven dominant negative ER mutants.
2)Using an adenovirus vector that carries PRL promoter-driven luciferase reporter gene, infection conditions in pituitary cultures were addressed. A maximal infection efficiency was obtained by infection at 5 MOI during the early culture period in the presence of insulin. Even at this infection efficiency, the proportion of reporter gene-expressing cells in lactotrophs was no more than 30%.
Research project 3
1)We produced several lines of transgenic rats carrying PRL promoter-driven GFP gene.
2)Among the transgenic rat lines, we chose MI-11-90 line on the basis of the fluorescence intensity, the proportion of GFP-expressing cells in lactotraphs, and the lactotroph-specificity for GFP expression.
3)Using FACS, we succeeded in selectively collect ting GFP-expressing pituitary cells, 90% of which were lactoarophs.
4)The stimulatory action of estrogen on lactotroph proliferation was enhanced in this lactotroph-enriched population while its inhibitory action of proliferation in the presence of IGF-1 was completely eliminated.
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