Approach to the mechanism of fertilization and reproductive medicine

受精机制与生殖医学探讨

• 批准号: 10470344
• 负责人: HOSHI Kazuhiko
• 金额: $ 4.03万
• 依托单位: Yamanashi Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470344/
• 关键词: fertilization; fertilizing ability; sperm; oocyte; progesterone; progesterone receptor; estrogen receptor; egg activation

1. The participation to the sperm fertilizing ability acquisition of sex steroid hormones and analysis of mechanisms for the process/Progesterone has been demonstrated to induce the acrosome reaction in the human spermatozoon via the cell surface membrane progesterone receptor (PR). However, the molecular identity of the membrane receptor has not been clarified yet. We identified a novel isoform (termed "isoform S") of the progesterone receptor mRNA (PR isoform S mRNA ,PR-S mRNA) which consisted of a previously unidentified 5' -untranslated sequence and the exon 4-8 of the intracellular PR gene from the human testicular cDNA library. The PR-S mRNA could encode the protein corresponding to the almost entire part of the hormone binding domain of the intracellular PR, termed "PR isoform S (PR-S)". The 5' -untranslated sequence of the message was confirmed to be derived from a novel exon (termed "exon S") by the genomic cloning. Moreover, the expression level of the PR-S mRNA was higher in … More the spermatozoon than that in the uterine endometrium.Estrogen induces the influx of Ca++ into sperm cell, and this action is estimated with non-genomic action via the cell surface membrane estrogen receptor (ER), mo. We identified a novel isoform ( termed "isoform S" ) of the estrogen receptor mRNA (ER is form S mRNA ,Era -S mRNA) which consisted of a previously unidentified 5' -untranslated sequence and the exon 4-8 of the intracellular ER gene from the human testicular cDNA library. The ERα -S mRNA could encode the protein corresponding to the almost entire part of the hormone binding domain of the intracellular ER, termed "Era isoform S (ERα - S).These results implied that the PR-S and ERα -S which were possibility expressed in the spermatozoon might be related to the cell surface membrane steroid hormone receptors.2. Effect of sperm immobilization and demembranation on the oocyte activation rate in the mouseTo analyze the effect of the state of sperm membrane on the oocyte activation rate following intracytoplasmic sperm injection (ICSI), three types of human and mouse spermatozoa (intact, immobilized and Triton X-100 treated) were individually injected into mouse oocytes.Following human sperm injection, the fastest and the most efficient oocyte activation and sperm head decondensation occurred when the spermatozoa were treated with Triton X-100. Intact spermatozoa were the least effective in activating oocytes. When mouse spermatozoa were injected into mouse oocytes, the rates of oocyte activation and sperm head decondensation within activated oocytes were the same irrespective of the types of sperm treatments prior to injection. Less

1. 性类固醇激素参与精子受精能力的获得及其机制分析/孕酮通过细胞膜孕酮受体（PR）在人精子中诱导顶体反应。然而，膜受体的分子特性尚未明确。我们从人类睾丸cDNA文库中鉴定了一个新的孕激素受体mRNA （PR isoform S mRNA,PR-S mRNA）的异构体（称为“异构体S”），该异构体由先前未识别的5' -未翻译序列和细胞内PR基因的4-8外显子组成。PR-S mRNA可以编码细胞内PR几乎全部激素结合域对应的蛋白，称为PR异构体S （PR-S）。通过基因组克隆证实，该信息的5'未翻译序列来源于一个新的外显子（称为“外显子S”）。PR-S mRNA在精子中的表达量高于子宫内膜。雌激素诱导Ca++流入精子细胞，这一作用是通过细胞表面膜雌激素受体（ER）的非基因组作用来估计的。我们从人类睾丸cDNA文库中鉴定了雌激素受体mRNA的一个新的异构体（称为“S异构体”）（ER是S型mRNA，Era -S mRNA），该异构体由先前未识别的5' -未翻译序列和细胞内ER基因的4-8外显子组成。ERα -S mRNA可以编码几乎整个细胞内内质网激素结合域对应的蛋白，称为Era异构体S （ERα -S）。这些结果提示PR-S和ERα -S可能在精子中表达，可能与细胞膜类固醇激素受体有关。精子固定化和脱膜对小鼠卵母细胞活化率的影响为了分析精子膜状态对卵浆内单精子注射（ICSI）后卵母细胞活化率的影响，分别将完整、固定化和Triton X-100处理的3种人和小鼠精子注射到小鼠卵母细胞中。在人类精子注射后，用Triton X-100处理精子时，卵母细胞激活和精子头去致密的速度最快，效率最高。完整精子激活卵母细胞的效果最差。将小鼠精子注射到小鼠卵母细胞中，无论注射前的精子处理方式如何，激活卵母细胞的卵母细胞活化率和激活卵母细胞内精子头部去浓缩率都是相同的。少

项目成果

Kato J et al.: "The multiple untranslated first exon and promotor system of the oestrogen receptor gene in the brain and peripheral tissues of the rat and the developing rat cerebral cortex."J Steroid Biochem Molec Biol. 65. 281-293 (1998)

Kato J 等人：“大鼠大脑和外周组织以及发育中的大鼠大脑皮层中雌激素受体基因的多个非翻译第一外显子和启动子系统。”J Steroid Biochem Molec Biol。

Kasai T et al.: "Fixed schedule for in vitro fertilization and embryo transfer: comparison of outcome between the short and the long protocol."Yamanashi Med J. 14. 77-82 (1999)

Kasai T 等人：“体外受精和胚胎移植的固定时间表：短方案和长方案之间结果的比较。”Yamanashi Med J. 14. 77-82 (1999)

Mizuno K et al.: "Analysis of the phospholipid hydroperoxide glutathione peroxidase mRNA In the rat spermatozoon and effect of selenium deficiency on the mRNA."Biol Trace Element Res. 74(In press).

Mizuno K 等人：“大鼠精子中磷脂氢过氧化物谷胱甘肽过氧化物酶 mRNA 的分析以及缺硒对 mRNA 的影响。”Biol Trace Element Res。

Kasai, T: "Fixed schedule for in vitro fertilization and embryo transfer: comparison of outcome between the short and the long protocol."Yamanashi Med J. 14,3. 77-82 (1999)

Kasai, T：“体外受精和胚胎移植的固定时间表：短期和长期方案之间的结果比较。”Yamanashi Med J. 14,3。

大田昌治: "プロゲステロンの精子受精能に与える効果"日本受精着床学会雑誌. 16,1. 85-88 (1999)

Shoji Ota：“黄体酮对精子受精能力的影响”日本受精与着床学会杂志 16,1（1999）。