| Project/Area Number |
10660276
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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 |
Applied animal science
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| Research Institution | Nippon Veterinary and Animal Science University |
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Principal Investigator |
TOMOGANE Hiroshi Nippon Vet & Anim Sci Univ Professor, 獣医畜産学部, 教授 (30023493)
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| Co-Investigator(Kenkyū-buntansha) |
OHBOSHI Shigeki Nippon Vet & Anim Sci Univ Lecturer, 獣医畜産学部, 講師 (10277668)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | mouse fertilized ovum / collagen / cis-4-hydroxy-2-proline / collagen gene / morula / blastocyst / 初期胚発育 / ラット / ウシ胎子血清 / インシュリン / アクチン / 受精卵発育 / 胚体外発生培養 / コラーゲンタイプ / マウス / 胚の初期発育 / ヒドロキシプロリン異性体 / 細胞外マトリックス / マウス胚 |
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| Research Abstract |
Study(1) : A fertilized ovum repeats cell divesion cycles to increase the number of cell. The increased cell forms morula and blastocyst. In all tissues there is always collagen as extracellular matrix. To determine the contribution of collagen to the development of preimplantation embryos, cis-4-hydroxy-2-proline (CHP), collagen sysnthesis inhibitor, was added into the medium for the in vitro development of mouse 2-cell embryo and the in vitro development of 2-cell stage to blastocyst was estimated by the developmental rate to 4-cell, 8-cell, morula and blastocyst and by the number of cell at the developed blastocyst. The CHP addition blocked to develop to 4-cell satge from 2-cell stage. This blocking effect was inhibited by the replacement of proline at the same of CHP addition but not the replacement of lysine or glycine. Study(2) : The blocking effect of CHP on in vitro embryonic development was examined at various stages of embryos. The blocking occurred remarkably at the stages of from 2-cell to 4-cell and from morula to blastocyst. Study(3) : Using reverse transeription polymerase chain reaction method, mRNA of type I and III collagen weakly appeared at the stage of 4-cell. The typelV collagen mRNA first appeared from 8-cell stage and more at the blastocy. Study(4) : The collagen at the various stages of preimplantation embryos was determined using immuno-fluoresence- histochemistry. The collagen immuno-reaction was weakly detected at 2-cell, 4-cell and 8-cell stages and strongly at both morula and blastocyst cmbryos. These results indicate that collagen has important role for the early embryonic development and is essential subustance to develop to 4-cell stage and to blastocyst from morula.
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