Functional analysis of germ line cells using gene manipulated mouse

使用基因操作小鼠进行生殖系细胞的功能分析

• 批准号: 11234203
• 负责人: OKABE Masaru
• 金额: $ 54.4万
• 依托单位: OSAKA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11234203/
• 关键词: gene manipulate animals; transgenic mouse; GFP; sex differentiation; sex chimera; ACE; GPI-anchor; 生殖細胞; 複数分裂; 雌雄キメラ; Cre; 1oxP; loxP; 雌雄選別; キメラ

We have developed an easy and very efficient system to separate male and female embryos before implantation stage with non invasive manner. When male and female eggs prepared by this system were aggregated to produce sex chimera, the advantage of this system is the easiness of pointing out the sex of individual cells that form organs by observing the transgenically incorporated green fluorescent protein gene in X chromosome. By analyzing the genital organs in thus prepared sex chimeras, we have found many spermatogonia like cells of female origin in phenotypically male testes. The pattern of the genomic imprinting of these genes was found out to be male type in spite of their original sex was female.With the collaboration to Prof. Nakatsuji's group, it was also found out that these "female spermatogonia" cells entered meiosis in early stages of development However, on the other hand, some cells originated from female became huge cells like eggs in the testis of sex chimera animals, whi … More le the majority of the female cells developed into male type spermatogonia. The complexity of the sex differentiation of germ cells was elucidated by producing these sex chimera mice and simultaneously provided a tool to analyze the mechanism of male-female differentiation in germ line cells.Pig-a, an X-linked gene, is a key component of glycosylphosphatidylinositol (GPI) anchor biosynthesis based on the fact that lack of this gene causes deficiencies of hundreds of GPI-anchored proteins. Dr. Kondoh has produced a transgenic mouse line that expressed enhanced green fluorescent protein in GPI-anchored form and monitored GPI-anchor-positive cells in situ. It was found out that ACE has phospholipase activity other than the dipeptidyl-peptidase. It was suggested that testicular ACE is indispensable because it may release GPI anchored proteins from sperm surface to make fertile sperm. The results indicated that the necessity of re-evaluation of the involvement of ACE in reproductive biology. Less

我们开发了一种简单而有效的系统，可以在植入前以非侵入性方式分离男性和女性胚胎。当通过该系统制备的雄卵和雌卵聚集以产生性嵌合体时，该系统的优点是通过观察X染色体中转基因掺入的绿色荧光蛋白基因，容易指出形成器官的单个细胞的性别。通过对制备的性嵌合体生殖器官的分析，我们发现在表型为雄性的睾丸中有许多雌性起源的精原细胞样细胞。在与Nakatsuji教授的研究小组的合作下，发现这些基因的基因组印迹模式是雄性型的，尽管它们的原始性别是雌性。这些“雌性精原细胞”在发育的早期阶段就进入减数分裂，但另一方面，一些来自雌性的细胞在性嵌合体动物的睾丸中变成了巨大的细胞，如卵子，这是一个重要的研究领域。 ...更多信息 雌性细胞大多发育为雄性型精原细胞。Pig-a是一个X连锁基因，是糖基磷脂酰肌醇（GPI）锚合成的关键组分，该基因的缺失导致数百种GPI锚定蛋白的缺失。Kondoh博士已经产生了一种转基因小鼠系，其以GPI锚定形式表达增强的绿色荧光蛋白，并原位监测GPI锚定阳性细胞。发现ACE具有磷脂酶活性而不是二肽基肽酶。提示睾丸ACE可能是从精子表面释放GPI锚定蛋白，使精子具有生育力的重要途径。提示有必要重新评价ACE在生殖生物学中的作用。少

项目成果

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