Expression and role of exocytotic SNARE proteins on oocyte cortical reaction

胞吐 SNARE 蛋白在卵母细胞皮质反应中的表达及其作用

• 批准号: 11671649
• 负责人: KUJI Naoaki
• 金额: $ 1.98万
• 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: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671649/
• 关键词: oocytes; spermatozoa; testis; exocytosis; syntaxin; vesicle-associated membrane protein (VAMP)

Exocytotic SNARE protein syntaxin expression was confirmed in both mouse oocytes and ovarian tissue, through SDS-PAGE using anti-ratsyntaxin 1B rabbit polyclonal antibody we raised previously. We also confirmed the existence of syntaxin 4 mRNA in total RNA from mouse oocytes by RT-PCR analysis, and cDNA sequence was completely coincides with mouse syntaxin 4.Localization of this syntaxin 4 like molecule was examined by confocal microscopy, and it was revealed that syntaxin antigenesity was co-localized with cortical granule distribution before and after cortical reaction. After this observation, it was supposed syntaxin 4 might play some functional role during mouse oocyte cortical reaction as a component of fusion machinery.Syntaxin was supposed to exist on acrosomal region of human spermatozoa form immunocytochemistry and SDS-PAGE analysis, suggested to engage sperm acrosome reaction.Also, another exocytotic protein VAMP was expresseed on memmbrane fraction from rat testis. This VAMP-like protein was expressed on spermatogonia in testis before puberty in rat, although its functional role remained tobe elucidated.

用我们制备的抗ratsyntaxin 1B的兔多克隆抗体进行SDS-PAGE，证实了SNARE蛋白syntaxin在小鼠卵母细胞和卵巢组织中均有表达。RT-PCR分析证实小鼠卵母细胞总RNA中存在syntaxin 4 mRNA，其cDNA序列与小鼠syntaxin 4完全一致。共聚焦显微镜观察syntaxin 4样分子的定位，发现syntaxin抗原的表达与皮质反应前后皮质颗粒的分布存在共定位关系。结果表明，syntaxin 4可能作为融合机制的一个组成部分，在小鼠卵母细胞皮质反应中发挥一定的功能;免疫细胞化学和SDS-PAGE分析表明Syntaxin存在于人精子顶体区，参与精子顶体反应;大鼠睾丸膜上也表达有一种胞吐蛋白VAMP。这种VAMP样蛋白在青春期前的大鼠睾丸精原细胞上有表达，但其功能尚不清楚。

项目成果

田中宏明,久慈直昭 他: "ラット雄性配偶子系列における開口放出関連蛋白の発現"日本不妊学会雑誌. 45. 568 (2000)

Hiroaki Tanaka、Naoaki Kuji 等：“大鼠雄性配子谱系中胞吐作用相关蛋白的表达”日本不孕协会杂志 45. 568 (2000)。

Iwahashi K, Kuji N, Tanaka H, Kitaoka Y, Shiraishi S, Asada H, Hamatani T, Ozawa N, Miyazaki T, Sueoka K, Yoshimura Y, Akagawa K.: "Expression of HPC-1/syntaxin mRNA on murine oocytes during oocyte maturation and early embryo development."Jpn J Fertil Ste

Iwahashi K、Kuji N、Tanaka H、Kitaoka Y、Shiraishi S、Asada H、Hamatani T、Ozawa N、Miyazaki T、Sueoka K、Yoshimura Y、Akakawa K.：“HPC-1/突触蛋白 mRNA 在小鼠卵母细胞上的表达

Tanaka H, Kuji N, Tsuchiya S, Tanigaki R, Hamatani T, Ozawa N, Miyazaki T, Sueoka K, Yoshimura Y, Fujiwara T, Akagawa K.: "Expression of exocytotic SNARE protein on rat male germ cell lineage."Jpn J Fertil Steril. 45 (4). 568 (2000)

Tanaka H、Kuji N、Tsuchiya S、Tanigaki R、Hamatani T、Ozawa N、Miyazaki T、Sueoka K、Yoshimura Y、Fujiwara T、Akakawa K.：“大鼠雄性生殖细胞谱系中胞吐 SNARE 蛋白的表达。”Jpn J

岩橋和裕,久慈直昭 他: "マウス卵における開口放出関連蛋白HPC1/syntaxin の存在について"日本産科婦人科学会誌. 50. S155 (1998)

Kazuhiro Iwahashi、Naoaki Kuji 等：“小鼠卵中胞吐作用相关蛋白 HPC1/syntaxin 的存在”日本妇产科学会杂志 50. S155 (1998)。

岩橋和裕,久慈直昭 他: "マウス各成熟期卵子におけるsyntaxin4mRNAの発現と解析"日本不妊学会雑誌. 45. 563 (2000)

Kazuhiro Iwahashi、Naoaki Kuji 等：“不同成熟阶段小鼠卵母细胞中突触蛋白 4 mRNA 的表达和分析”日本不孕协会杂志 45. 563 (2000)。