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Apoptosis Uterine Endometrium

子宫内膜凋亡

基本信息

批准号：
08671939
负责人：
OTSUKI Yoshinori
金额：
$ 1.6万
依托单位：
Osaka Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671939/
关键词：
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

项目摘要

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 … More 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

bcl-2缺陷小鼠的子宫的特征是生长迟缓，包括缺乏腺体分叉和固有层和子宫肌层发育不良。因此，bcl-2是子宫内膜腺细胞和子宫肌层平滑肌细胞在子宫中存活的必需基因（Daikoku et al.，在本研究中，我们开发了两种检测DNA链断裂的技术：DNA链断裂的原位末端标记应用透射电子显微镜（TEM）和共聚焦激光显微镜（CLM）对人子宫内膜腺细胞进行了iISEL技术研究，观察了人子宫内膜腺细胞在月经周期中DNA片段化的变化。使用NIH图像程序在计算机上进行DNA链断裂定量变化的图像分析。此图像 ...更多信息通过TEM技术结合ISEL的ge分析清楚地表明，在晚期增殖阶段，当细胞在形态上仍被保留时，子宫内膜腺细胞核中DNA链断裂的标记密度增加（早期增殖阶段：23.4（μ S·± 1）μ S·± 1）。D_1期9.1个/μ m，增殖晚期142.6个/μ m。D12 @>D1），凋亡小体标记密度最高（325.4 ± 0.01）。[）166.1/mum<@D12@>D1）。因此，结合ISEL/TEM技术的图像分析允许我们半定量细胞核中的DNA链断裂，并且它提供了关于凋亡细胞的形态学变化与细胞核中DNA链断裂的标记密度变化之间的关系的有用信息（Inoki等人，利用CLM的iISEL技术，通过重金属产物的反射率检测游离3 '-OH DNA末端的位点，并通过甲基绿色的自发荧光证明双链DNA的定位。通过CLM的ISEL技术清楚地显示了游离3 '-OH DNA末端和双链DNA的同时定位，证明其具有广泛的应用，包括其他DNA自溶过程的研究（Ito等人，J.Histochem.细胞化学，印刷中）。少