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Attempts to establish a screening procedure for endocrine disrupters by ovulation analysis using the intrabursal injection method

尝试通过囊内注射法进行排卵分析来建立内分泌干扰物筛查程序

基本信息

批准号：
11554036
负责人：
TAKAHASHI Takayuki
金额：
$ 7.94万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11554036/
关键词：
vertebrate ovulation intrabursal injection method endocrine disrupter

项目摘要

he aim of this study was to establish the basis for a acreening method of endocrine disrupters using intrabursal injection analysis of mammalian ovulation.We first attempted to search for molecular markers suitable for this method. To this end, bradykinin B2 receptor and MMP-23(a new type of matrix metalloproteinase)were examined whether they would be appropriate as markers. The B2 receptor mRNA and receptor protein were both detected in the ovaries of pig and mouse. Further detailed morphological examination revealed that the B2 receptor mRNA was constitutively expressed in the granulosa cells of all growing follicles in the ovary, and that the mRNA levels did not change through the periovulatory period. These findings indicate that the B2 receptor may not serve as an appropriate marker. The expression of MMP-23 was examined using rat ovaries for the same purpose. MMP-23 mRNA was localized at granulosa cells of growing follicles in the early stages of follicle growth, but its expression was shifted to the outer thecal layer of fully grown follicles. Based on these results, we presumed that MMP-23 could be a marker for follicle growth rather than for ovulation.Effects of two known endocrine disrupters, DES and DEHP, on ovulatory process were examined by applying for the ovulation analysis method using female rats and mice. Intrabursal injection of DES and DEHP into rats did not resulted in significant decrease in the numbers of ovulated eggs. However, female offsprings of the DES-treated mother mice ovulated substantially reduced numbers of eggs. These results indicate that DES treatment affected profoundly reproductive ability of the offspring.

本研究的目的是建立一种利用囊内注射分析哺乳动物排卵的内分泌干扰物筛选方法的基础，我们首先尝试寻找适合这种方法的分子标记。为此，研究缓激肽B2受体和MMP-23（一种新型基质金属蛋白酶）是否适合作为标志物。在猪和小鼠卵巢中均检测到B2受体mRNA和受体蛋白。进一步详细的形态学检查显示，B2受体mRNA在卵巢中所有生长卵泡的颗粒细胞中组成型表达，并且mRNA水平在排卵期没有变化。这些发现表明，B2受体可能不是一个合适的标记。为了同样的目的，使用大鼠卵巢检测MMP-23的表达。MMP-23 mRNA在卵泡生长早期定位于生长卵泡的颗粒细胞，而在完全生长卵泡的卵泡膜外层表达。本研究应用大鼠和小鼠排卵分析方法，观察了两种已知的内分泌干扰物DES和DEHP对排卵过程的影响。囊内注射己烯雌酚和邻苯二甲酸二异辛酯对大鼠的排卵数无明显影响。然而，DES处理的母鼠的雌性后代排卵的卵子数量大幅减少。这些结果表明，DES处理对后代的繁殖能力产生了深刻的影响。