Molecular biology of species-specific sperm and zona pellucida interactions

物种特异性精子与透明带相互作用的分子生物学

• 批准号: 14370536
• 负责人: KOYAMA KOJI
• 金额: $ 6.21万
• 依托单位: Hyogo College of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370536/
• 关键词: zona pellucida; transgenic mouse; ZP2- knockout mouse; pig ZPA; oocyte; fertilization; infertility; ovarian follicular disorder; RNA干渉; 卵透明帯; 卵胞; 顆粒膜細胞; ZPA; ZPB; ZPC; 精子; ノックアウトマウス; 不妊症; トランスジェニック

The zona pellucida, an extracellular matrix surrounding mammalian oocytes, is composed of three glycoproteins designated as ZP1(ZPB), ZP2(ZPA) and ZP3(ZPC). During fertilization, zona pellucida glycoproteins play several critical roles such as recognition of sperm, induction of the acrosome reaction and prevention of polyspermy. The purpose of this study was to examine the molecular mechanism of expression and incorporation of pig zona pellucida glycoprotein 2 (pZP2) in mouse zona pellucida. A transgene construct was prepared by placing a minigene that encodes pig ZP2 downstream from the promoter of mouse ZP2. Using this construct, we obtained four mouse lines harboring the pig ZP2 gene. The transgenic protein was found to be synthesized only in growing oocytes and expressed on the surface of the oocytes within the ovary. However, it was not detected in ovulated eggs. Furthermore, the pZP2 transgene did not rescue the phenotype in ZP2-knockout zona-deficient mice. These results indicated that in transgenic mice pig ZP2 is synthesized but not incorporated into the mouse zona pellucida, possibly due to failure to combine with other mouse ZP components (ZP1 and ZP3). It is therefore concluded that the zona pellucida is constituted from the component proteins in a molecular species-specific manner.

透明带是哺乳动物卵母细胞周围的一种细胞外基质，由三种糖蛋白组成，分别为ZP1(ZPB)、ZP2(ZPA)和ZP3(ZPC)。在受精过程中，透明带糖蛋白在识别精子、诱导顶体反应和防止多精受精等方面发挥着重要作用。本研究旨在探讨猪卵透明带糖蛋白2(PZP2)在小鼠卵透明带中表达和掺入的分子机制。将编码猪ZP2的微型基因置于小鼠ZP2启动子下游，构建了转基因载体。利用该构建物，我们获得了4个含有猪ZP2基因的小鼠株系。转基因蛋白仅在发育中的卵母细胞中合成，并在卵巢内的卵母细胞表面表达。然而，在排卵的卵子中没有检测到它。此外，pZP2转基因不能挽救ZP2基因敲除小鼠的表型。这些结果表明，在转基因小鼠中，猪ZP2被合成，但没有整合到小鼠的透明带中，这可能是由于未能与其他小鼠ZP组分(ZP1和ZP3)结合所致。因此，可以得出结论，透明带是由组成蛋白以分子物种特有的方式构成的。

项目成果

Pig zona pellucida 2 (pZP2) protein does not participate in zona pellucida formation in transgenic mice.

猪透明带 2 (pZP2) 蛋白不参与转基因小鼠透明带的形成。

• 发表时间: 2006
• 期刊: Reproduction
• 影响因子: 3.8
• 作者: A. Hasegawa;Nozomi Kanazawa;H. Sawai;S. Komori;K. Koyama
• 通讯作者: K. Koyama

Contribution of zona proteins to oocyte growth.

• 发表时间: 2007
• 期刊: Society of Reproduction and Fertility supplement
• 作者: A. Hasegawa;K. Koyama