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Studies on the Regulation of Fertilization and Activation of Development

受精调控与发育激活研究

基本信息

批准号：
12306018
负责人：
BABA Tadashi
金额：
$ 28.24万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12306018/
关键词：
Sperm Egg Fertilization Sperm maturation Egg activation Cell fusion Oviduct Knockout mouse 卵活性化

项目摘要

To elucidate the regulation of fertilization and activation of development, we have examined possible functions of mouse sperm in sperm maturation, sperm transit from uterus to oviduct, fertilization, and egg activation. The following experimental results have been obtained :1. The TritonX-100-insoluble microdomains, lipid rafts, on the plasma membranes of epididymal sperm gradually move to the apical region from the equatorial region of sperm during capacitation.2. Male mice lacking sperm PH-20 are still fertile, in spite of a reduced ability to disperse cuwnulus cells from the cumulus mass. Western blot analysis of sperm extracts reveals the presence of other hyaluronidase(s), except PH-20, presumably within the sperm acrosome. These data provide evidence that PH-20 is not essential for fertilization at least in the mouse, and suggest that the other hyaluronidase(s) may play an important role in the sperm penetration through the cumulus cell layer and/or the egg zona pellucida, possi … More bly in cooperation with PH-20.3. Mouse genomic clones encoding a 42-kDa sperm serine protease, TESP5, have been identified. Immunochemical analysis reveals that 42- and 4l-kDa forms of TESP5 are localized in the head, cytoplasmic droplet, and tail of cauda epididymal sperm probably as a glycosylphosphatidylinositol-anchored membranous protein. Moreover, these two forms of TESP5 are selectively included into Triton X-100-insoluble microdomains, Iipid rafts, of the sperm membranes. These results suggest a possible involvement of TESP5 in the sperm/egg interactions.4. Two different ADAMI isoforms, ADAMla and ADAMlb, instead of ADAMl are present in the mouse testis. ADAMla is resttrictedly present within the endoplasmic reticulum of germ cells, whereas epididymal sperm contain only ADAMlb on the cell surface. The precursors of ADAMla and ADAMlb form a heterodimeric complex with that of ADAM2 in the endoplasmic reticulum of germ cells. The heterodimeric complex between the mature forms of ADAMlb and ADAM2 is also found on the sperm surface. These data imply the potential roles of ADAMla and ADAMlb in spermatogenesis and fertilization, respectively.5. When a synthesized mRNA with a poly(A) tail encoding phospholipase ζ is injected into unfertilized eggs, pronuclear formation is observed. Less

为了阐明受精和激活发育的调节，我们研究了小鼠精子在精子成熟、精子从子宫到输卵管的运输、受精和卵子激活中的可能功能。取得了如下实验结果：1.精子获能过程中，精子质膜上的TritonX-100不溶性微区（脂筏）逐渐从赤道区向顶区移动.缺乏精子PH-20的雄性小鼠仍然具有生育能力，尽管从卵丘群中分散卵丘细胞的能力降低。精子提取物的蛋白质印迹分析显示，除PH-20外，其他透明质酸酶可能存在于精子顶体中。这些数据提供了证据，表明PH-20至少在小鼠中不是受精所必需的，并表明其他透明质酸酶可能在精子穿透卵丘细胞层和/或卵透明质层中起重要作用，可能是因为它们在精子穿透卵丘细胞层和/或卵透明质层中起重要作用。 ...更多信息与PH-20.3配合使用。已经鉴定了编码42-kDa精子丝氨酸蛋白酶TESP 5的小鼠基因组克隆。免疫化学分析表明，42-和41-kDa形式的TESP 5定位在头部，细胞质液滴，尾部附睾精子可能作为糖基磷脂酰肌醇锚定的膜蛋白。此外，这两种形式的TESP 5被选择性地包含在精子膜的Triton X-100不溶性微区，脂筏中。这些结果表明TESP 5可能参与精子/卵子的相互作用.在小鼠睾丸中存在两种不同的ADAM 1亚型ADAM 1a和ADAM 1b，而不是ADAM 1。ADAM 1a限制性地存在于生殖细胞的内质网中，而附睾精子仅在细胞表面上含有ADAM 1b。ADAM 1a和ADAM 1b的前体在生殖细胞的内质网中与ADAM 2的前体形成异二聚体复合物。成熟形式的ADAM 1b和ADAM 2之间的异二聚体复合物也存在于精子表面。这些数据表明ADAM 1a和ADAM 1b分别在精子发生和受精过程中具有潜在的作用.当一个合成的mRNA与poly（A）尾巴编码磷脂酶α注入未受精卵，原核形成观察。少