Study on the mechanisms of oocyte-sperm interaction in fertilization process

受精过程中卵母细胞与精子相互作用的机制研究

• 批准号: 09671714
• 负责人: SUEOKA Kou
• 金额: $ 1.79万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671714/
• 关键词: Cell-free decondensing system; disulfide bond; lysolecithin; dithioshreitol; histon H2B; protamine; tapering sperm; nucleoplasmin; cell-free脱凝縮; Triton X-100; Xenopus laevis; protamine

Sperm nuclear proteins were studied to reveal the process of human sperm decondensation. Using in vitro decondensing system sperm nuclear transformation were clearly observed. This decondensing system was 0.05% lysolecithin - 5mM dithiothreitol - 1.OM NaCl solution. Most of sperm nuclei were decondensed completely in this system. Basic nuclear proteins were extracted from decondensed nuclei with 35% acetic acid or O.4N H_2S0_4, and were separated on SDS-PRGE and transferred onto a nitrocellulose membrane. Following incubation with anti-human histone H2B antibody and HRP-labelled secondary antibody, a blot was observed. This blot exhibited mobilities of 66 kDa. Furthermore another experiment showed the existence of histone H2B.Using anti- histone H2B antibody and FITC-labelled secondary antibody, the morphological and fluorescent changes of gradually decondensing sperm nuclei were observed on a glass slide. These results suggested that human sperm nuclei contain histones considerably after nucleoproteins were replaced by protamines in the process of sperm nuclear maturation.

研究了精子核蛋白，揭示了人类精子去浓缩的过程。用体外脱密系统可以清楚地观察到精子的核转化。该脱密体系为0.05%溶卵磷脂- 5mM二硫苏糖醇- 1。OM NaCl溶液。在这个系统中，大多数精子核完全去致密。用35%醋酸或0.4 n H_2S0_4从去浓缩核中提取碱性核蛋白，SDS-PRGE分离后转移到硝化纤维素膜上。用抗人组蛋白H2B抗体和hrp标记的二抗孵育后，观察印迹。该印迹的迁移率为66 kDa。另一项实验证实了组蛋白H2B的存在。用抗组蛋白H2B抗体和fitc标记的二抗，在玻片上观察逐渐去致密的精子核的形态和荧光变化。这些结果表明，在精子核成熟过程中，核蛋白被精蛋白取代后，人类精子核中含有大量组蛋白。

项目成果

黒島正子: "初期胚における染色体異常の発生頻度と胚発生過程でのembryo lossに関する一考察" 産婦人科の実際. 47. 1199-1203 (1998)

Masako Kuroshima：“早期胚胎染色体异常频率和胚胎发育过程中胚胎丢失的研究”妇产科实践 47. 1199-1203 (1998)。

K.Sueoka: "Integrins and reproductive physiology: expression and modulation in fertilization, embryogenesis, and implantation" Fertility & Sterility. 67(5). 799-811 (1997)

K.Sueoka：“整合素和生殖生理学：受精、胚胎发生和植入中的表达和调节”

岩田壮吉: "卵細胞質抽出物によるcell-free系精子核脱凝縮モデルの検討" 日本受精着床学会雑誌. 15(1). 98-101 (1998)

Sokichi Iwata：“使用卵细胞质提取物的无细胞精子核解凝模型的研究”日本受精与着床学会杂志 15(1) (1998)。

末岡 浩: "初期胚の遺伝子診断" 産婦人科の世界. 49(7). 35-43 (1997)

Hiroshi Sueoka：“早期胚胎的基因诊断”《妇产科世界》49(​​7) (1997)。

末岡 浩: "新女性医学大系" 中山書店(印刷中), (1998)

末冈浩：《女性医学的新体系》中山书店（正在出版），（1998）