Morphological studies of spermatozoa during capacitation and in relation to attachment mechmism with zona pellucida.

获能过程中精子的形态学研究以及与透明带附着机制的关系。

• 批准号: 03670004
• 负责人: TOYOTA Fumie
• 金额: $ 1.22万
• 依托单位: CHIBA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670004/
• 关键词: capacitation; boar spermatozoa; freeze-fracture; mouse spermatozoa; zona pellucida recognition and binding protein; immuno-cytochemistry; confocal laser scanning microscope; labeled surface replica; SP56; 抗SP56モノクローナル抗体; 金コロイド標識レプリカ法; 膜内粒子; 膜ス

(1) Changes of the plasma membtane of boar spermatozoa durin capacitation : The distribution change of intramembranous particles (IMP) in the periacrosomal plasma membrane (PAPM) of the boar spermatozoa was studied by freeze-fracture, During capacitation, IMP and filipin-sterol-complex free areas expanded. Statistical analysis of the IMP density suggested that small IMPs largely disappeared, while the majority of the large IMPs persisted in the PAPM.These results suggest that the small particles and the filipin reactive sterols act as membrane stavilizers to prevent a precocious acrosome reaction, and have to ve temoved furing capacitation.(2) Distribution of egg recognition and binding protein, sp56, on the mouse spermatozoa : SP56 has stecific affinity for ZP3, one of the three glycoproteins constituting the zona pillucida( Bleil and Wassarman, 1990). In collaboration with the Dr. Bleil's Lab, we demonstrated by immuno-cytochemical study using a confocal microscope that sp56 molecules are localized on the crescent dorsal edge of the ouse spermatozoon, where is well corresponded with the reported ZP3 binding site. In EM level, we debeloped a method it has been demonstrated that the sp56 is absent in the tip of the hooked-shaped mouse sperm-head, and present in the outer surface of the plasma memvrace covering the dorsal edge.

(1)猪精子获能过程中质膜的变化：用冷冻断裂法研究了猪精子顶体周围质膜（PAPM）膜内颗粒（IMP）的分布变化。对IMP密度的统计分析表明，小IMP大部分消失，而大IMP大部分保留在PAPM中。这些结果表明，小颗粒和菲律宾反应性甾醇作为膜稳定剂，以防止过早的顶体反应，并在获能过程中被移除。(2)卵识别和结合蛋白sp 56在小鼠精子上的分布：sp 56对ZP3具有特异性亲和力，ZP3是构成卵透明质酸的三种糖蛋白之一（Bleil和Wassarman，1990）。我们与Bleil博士的实验室合作，通过共聚焦显微镜的免疫细胞化学研究证明，sp56分子定位于乌塞精子的新月形背侧边缘，与报道的ZP3结合位点相对应。在电镜水平上，我们建立了一种方法，证明了sp56在钩状小鼠精子头的尖端不存在，而存在于覆盖背缘的质膜外表面。

项目成果

Haneji T, Nagano T, Koide S S: "Identification of pyruvate carboxylase in 3T3-L1 cells by transblot and its correlation with differentiation into adipocytes." Biochem Int. 25. 995-1001 (1991)

Haneji T、Nagano T、Koide S S：“通过转印迹鉴定 3T3-L1 细胞中的丙酮酸羧化酶及其与分化为脂肪细胞的相关性。”

M.Maekawa,T.Nagano,K.Kamimura,T.Murakami,H.Ishikawa&M.Dezawa: "Distribution of actin‐filament bundles in myoid cells,Sertoli cells,and tunica albuginea of rat and mouse testes." Cell Tiss.Res. 266. 295-300 (1991)

M.Maekawa、T.Nagano、K.Kamimura、T.Murakami、H.Ishikawa 和 M.Dezawa：“大鼠和小鼠睾丸肌样细胞、支持细胞和白膜中肌动蛋白丝束的分布”。 .研究266。295-300（1991）

M.Maekawa&Y.Mishimune: "In vitro proliferation of germ cells and supporting cells in the neonatal mouse testis." Cell Tiss.Res.265. 551-554 (1991)

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