| Project/Area Number |
08671889
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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 |
Obstetrics and gynecology
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| Research Institution | Osaka University |
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Principal Investigator |
TOKUGAWA Yoshihiro Osaka University Medical School, Assistant Professor, 医学部, 助手 (70283786)
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| Co-Investigator(Kenkyū-buntansha) |
SIMOYA Kouitchirou Osaka University Medical School, Assistant Professor, 医学部, 助手 (40291950)
AZUMA Chihiro Osaka University Medical School, Lecturer, 医学部, 講師 (20151061)
SAJI Fumitaka Osaka University Medical School, Associate Professor, 医学部, 助教授 (30203897)
久保田 康愛 大阪大学, 医学部・附属病院, 医員
大橋 一友 大阪大学, 医学部, 助手
佐治 文隆 大阪大学, 医学部, 助教授 (90093418)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1997: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | oxytocin unresponsiveness / oxytocin binding capacity / N-glycosylation / regulatory element / 発現調節領域 / 糖鎖 / 発見調節領域 / ゲルシフトアッセイ |
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| Research Abstract |
1. We have established permnent cell-lines in which the gene for the human oxytocin receptor has been introduced into HeLa cells. These cells differ by the disruption of hte N-terminal N-glycosylation sites by site-directed mutagenesis of the transfected oxytocin receptor constructs. The mutation of N-glycosylation sites appear to affect neither their dissociation constant, nor affinites for various oxytocin related ligands. This suggests that the full glycosylation of oxytocin receptor observed in vivo is not essential for its activity.
2. We examined the localization and expression of the oxytocin receptor in human myometrium by means of in situ hybridization, immunohistochemistory, and Northern and Western blotting. In the term pregnant myometrium, oxytocin receptor was expressed in uterine smmoth muscles. The level of oxytocin receptor transcripts increased according to the course of pregnancy. We also found that the receptor protein was augmented at term and after the onset of labor.
3. We determined the nucleotide sequence of the 5'-flanking region of the mouse oxytocin receptor gene, and compared the sequence with human, rat, and bovine sequence. This comparison revealed that a region commencing at nt-1671 to -1581 was a clear homology among the species. This sequence information will help to characterize the regulatory components of oxytocin receptor.
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