| Project/Area Number |
08671939
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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 Medical College |
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Principal Investigator |
OTSUKI Yoshinori Osaka Medical College, Professor, 医学部, 教授 (50140166)
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| Co-Investigator(Kenkyū-buntansha) |
UEKI Minoru Osaka Medical College, Professor, 医学部, 教授 (40084892)
TUJIMOTO Yoshihide Osaka College, Professor, 医学部, 教授 (70132735)
ITO Yuko Osaka Medical College, Assistance Professor, 医学部, 講師 (40148432)
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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 | Apoptosis / Uterus / Bcl-2 / Bcl-2 protein / bcl-2 deficient mice / TUNEL method / Transmission electron microscopy / Confocal laser microscopy / Bcl-2 / Fas-ligand / Fas antigen / TUNEL |
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| Research Abstract |
The uteri of the bcl-2 deficient mice were characterized by growth retardation, including a lack of bifurcation of the glands and poorly developed lamina propria and myometrium. The most interesting finding was the presence of apoptotic cells in both the glandular epithelium and the myometrium.
Therefore, bcl-2 is an essential gene for the survival of both endometrial glandular cells and myometrial smooth muscle cells in the uteri (Daikoku et al., Medical Electron MIcroscopy, in press).
We developed the two techniques for the detection of DNA strand breaks during this research : in situ end labeling of DNA strand breaks (ISEL techniques) using transmission electron microscopy (TEM) and cofocal laser microscopy (CLM).
Changes of DNA fragmentation in human endometrial glandular cells throughout the mentrual cycle were studied via iISEL techniques using TEM.Moreover, image analysis for quantitative changes of DNA strand breaks was performed on a computer using the NIH Image program. This ima
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ge analysis combined with ISEL via TEM techniques clearly demonstrated the increase of the labeling density for DNA strand breaks in the nuclei of the endometrial glandular cells at the late proliferative phase when the cells were still preserved morphologically (early proliferative phase : 23.4 (]SY.+-。[) 9.1/mum<@D12@>D1 ; late proliferative phase : 142.6 (]SY.+-。[) 40.0/mum<@D12@>D1), and the most intense labeling density was seen in apoptotic bodies (325.4 (]SY.+-。[) 166.1/mum<@D12@>D1). Therefore, the image analysis combined with ISEL/TEM techniques permits us to semi-quantify DNA strand breaks in nuclei, and it provides useful information as to the relation between the morphological changes of apoptotic cells and changes of the labeling density for DNA strand breaks in the nuclei (Inoki et al., J.Histotechnology, 1997).
Using iISEL techniques via CLM,the site of free 3'-OH DNA ends was detected by the reflectance from heavy metal products, and the localization of double-stranded DNA was demonstarated by the autofluorescence of methyl green. The ISEL technique via CLM clearly showed the simultaneous localization of both free 3'-OH DNA ends and double-stranded DNA proved to have a wide range of applications including the study of other DNA autolytic processes (Ito et al., J.Histochem. Cytochem., in press). Less
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