Cytochemical study on the substances(ZP1-3, sp56, acrosin)working at mammalian fertilization.

哺乳动物受精作用物质（ZP1-3、sp56、顶体素）的细胞化学研究。

• 批准号: 08670006
• 负责人: TOYOTA Fumie
• 金额: $ 1.34万
• 依托单位: CHIBA UNIVERSITY,SCHOOL OF MEDICINE
• 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 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670006/
• 关键词: ZP3 receptor; sp56; aquaporin 7; spermatozoa; spermiogenesis; immuno-cytochemistry; laser confocal microscope; labeled surface replica; 水チャンネル; 細胞相互作用; 電子顕微鏡; マウス; ZP3受容体; 免疫細胞化学

1. Distribution of ZP3 receptor on the mouse spermatozoon : ZP3 is one of the three glycoproteins in the zona pellucida, and significant for the sperm recognition, binding and induction of acrosome reaction. The localization of ZP3 receptor on the spermatozoa was determined by colloidal gold-conjugated ZP3 and surface replica technique. The gold particles were detected on the outer surface of the plasma membrane covering the dorsal edge of the sperm head.2. Localization(1), molecular image (2), and formation (3) of sp56 :(1) sp56 has specific affinity for ZP3. By immuno- cytochemical and surface replica techniques, we demonstrated that sp56 localized at the same area, where the ZP3 receptor was demonstrated in 1. It suggests that sp56 satisfies conditions as a ZP3 receptor.(2) Molecular image of sp56 was observed by low angle rotary shadowing method. The octamer of sp56, which is the form in vivo, appeared as a four-leafed clover, having a maximum diameter of approximately 50nm. Each leaf is thought to be composed of a sp56 dimer.(3) During spermiogenesis, sp56 was observed in the acrosome. A question, when sp56 appear on the outer surface of the plasma membrane, remained to be answered.3. Formation and localization of aquaporin 7 (AQP7), a water channel protein, during spermiogenesis in the rat testis : Small size of spermatozoa is indispensable for penetration through the zona pellucida. Present immunohistochemical study demonstrated that AQP7 first appears in round spermatid. The plasma membrane covering the cytoplasmic mass showed strong immunoreactivity in the elongated spermatid. The result suggests that AQP 7 works as a water channel, since it is localized on the membrane covering the condensing and diminishing cytoplasm.

1. ZP3受体在小鼠精子上的分布：ZP3是透明质酸的三种糖蛋白之一，对精子识别、结合和诱导顶体反应具有重要意义。用胶体金标记的ZP3结合表面复制技术，对ZP3受体在精子上的定位进行了研究。在精子头部背缘的质膜外表面检测到金颗粒. sp56的定位（1）、分子图像（2）和形成（3）：（1）sp56对ZP3具有特异性亲和力。通过免疫细胞化学和表面复制技术，我们证明了sp56定位在相同的区域，在那里ZP3受体被证明在1。这表明sp56满足作为ZP3受体的条件。(2)用小角旋转阴影法观察了sp56的分子图像。sp56的八聚体是体内形式，表现为四叶三叶草，最大直径约为50 nm。每个叶片被认为是由一个sp56二聚体组成。(3)在精子发生过程中，sp56在顶体中被观察到。sp56何时出现在质膜外表面的问题尚待解答.水通道蛋白7（AQP7）在大鼠睾丸精子发生过程中的形成和定位：小尺寸的精子对于穿透透明膜是必不可少的。目前的免疫组化研究表明，AQP7首先出现在圆形精子细胞。长形精子细胞的质膜覆盖细胞质团，呈强免疫反应性。结果表明，AQP 7作为一个水通道，因为它是定位在膜覆盖的浓缩和缩小的细胞质。

项目成果

Suzuki F: "Modification of the mammalian sperm plasma membrane during spermiogenesis, maturation and capacitation.(in Japanese)" Seitai no Kagaku. 48. 72-79 (1997)

铃木 F：“精子发生、成熟和获能过程中哺乳动物精子质膜的修饰。（日语）”Seitai no Kagaku。

鈴木 二美枝 (豊田): "哺乳類精子の形成・成熟・受精能獲得" 生体の科学. 48(1). 72-79 (1997)

Fumie Suzuki（丰田）：“哺乳动物精子的形成、成熟和受精潜力的获得”《生物科学》48（1）（1997）。

Ishibashi K, Suzuki F, Sasaki S et al.: "Cloning and functional expression of a new water channnel expressed in the testis permeable to water…" The J.Biological Chemistry. 272. 20782-20786 (1997)

Ishibashi K、Suzuki F、Sasaki S 等人：“睾丸可渗透水的新水通道的克隆和功能表达……”J.Biological Chemistry 272. 20782-20786 (1997)

Suzuki F, Maekawa M, Bleil JD, et al.: "Localisation of a mouse sperm protein sp56, Which has specific affinity for a zona glycoprotein ZP3." Zygote. Suppl. s92- (1998)

Suzuki F、Maekawa M、Bleil JD 等人：“小鼠精子蛋白 sp56 的定位，该蛋白对透明带糖蛋白 ZP3 具有特异性亲和力。”

Suzuki F,Maekawa M,Bleil JD,et la.: "Localisation of a mouse sperm protein SP56,which has specific affinity for a zona glycoprotein ZP3." Zygote (Suppl). 1. s92 (1998)

Suzuki F、Maekawa M、Bleil JD 等人：“小鼠精子蛋白 SP56 的定位，该蛋白对透明带糖蛋白 ZP3 具有特异性亲和力。”