General principle involved in the mechanism of oocyte maturation

卵母细胞成熟机制的一般原理

• 批准号: 04044177
• 负责人: NAGAHAMA Yoshitaka
• 金额: $ 4.86万
• 依托单位: Okazaki National Research Institutes National Institute for Basic Biology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04044177/
• 关键词: Oocyte maturation; Vertebrate; Amphibians; Fishes; Maturation-promoting factor (MPF); Maturation-inducing hormone; Gonadotropin; リン酸・脱リン酸化; リン酸脱リン酸化

The purpose of this cooperative project was that two research groups directed by Drs.Yoshitaka Nagahama (Okazaki, Japan) and J.L.Maller (Denver, USA) collaborate to clarify the general principle involved in the regulatory mechanism of oocyte maturation in vertebrates, in particular amphibians (Maller, USA) and fishes (Nagahama, Japan).In both amphibians (Xenopus) and fishes (salmonids, goldfish, etc.), gonadotropins (probably LH-like gonadotropin) are responsible for triggering oocyte maturation. However, in both cases, gonadotropins do not act directly on the oocyte, but act on the ovarian follicle cells to produce maturation-inducing hormones (progesterone in amphibians and 17alpha, 20beta-dihydroxy-4-pregnen-3-one, 17alpha, 20beta-DP, in fishes). The site of the action of both maturation-inducing hormones is the surface of the oocytes ; specific surface receptors responsible for the progesterone-and 17alpha, 20beta-DP-induced oocyte maturation were found on the plasma membrane of Xe … More nopus, rainbow trout and flounder oocytes. Two research groups will continuously collaborate to purify and characterize these steroid hormone membrane receptors.Microinjection of progesterone and 17alpha, 20beta-DP failed to induce oocyte maturation, suggesting that they do not act directly on the germinal vesicle but seems to act through a third mediator in the oocyte cytoplasm. This third mediator of cytoplasmic nature has been called maturation-promoting factor (MPF). MPF was purified and characterized from oocytes of Xenopus and goldfish (carp), consisting of two common components, cdc2 kinase and cyclin B. Although the components of MPF in Xenopus and goldfish oocytes, are the same, the mechanisms of MPF activation are different between these two species. In goldfish, 17alpha, 20beta-DP induces immature oocytes to synthesize cyclin B, which in turn activates preexisting 35-kDa cdc2 kinase through its threonine (Thr161) phosphorylation, producing the 34-kDa active cdc2 kinase. These mechanisms of MPF activation in fish oocytes apparently differ from those in Xenopus, in which cyclin B is present in immature oocytes and forms a complex with cdc2 kinase (pre-MPF). Furthermore, in Xenopus oocytes, dephosphorylation of threonine (Thr14) and tyrosine (Tyr15) is required for cdc2 kinase activation. However, it is doubtful that tyrosine (Tyr15) dephosphorylation of cdc2 kinase is not a prerequisite for its activation during goldfish oocyte maturation, since the activation is not inhibited by vanadade, a protein phosphatase inhibitor commonly used for inhibiting cdc25 activity that dephosphorylates tyrosine (Tyr15) of cdc2 kinase, thereby inhibits its activation. Less

该合作项目的目的是，由Yoshitaka Nagahama（日本冈萨木）和J.L. -Maller（美国丹佛）指导的两个研究小组合作阐明了参与脊椎动物中卵母细胞成熟的监管机制涉及的一般原理（鲑鱼，金鱼等），促性腺激素（可能是LH样促性腺激素）负责触发卵母细胞的成熟。 However, in both cases, gonadotropins do not act directly on the oocyte, but act on the ovarian folicle cells to produce maturation-inducing horsemones (progesterone in amphibians and 17alpha, 20beta-dihydroxy-4-pregnen-3-one, 17alpha, 20beta-DP, in fishes).两种成熟骑马的作用部位是卵母细胞的表面。在Xe的质膜上发现了负责孕酮和17Alpha，20BETA-DP诱导的卵母细胞成熟的特定表面受体……更多的NOPUS，Rainbow Trout和Floater卵母细胞。两个研究小组将继续合作以净化和表征这些立体的骑马膜受体。孕酮和17Alpha的米切尔注射未能诱导卵母细胞成熟，这表明它们不直接作用于生发蔬菜，但似乎通过卵母细胞细胞质量的第三个介质作用。第三个细胞质性质的介体称为成熟因子（MPF）。 MPF被纯化并从异武和金鱼（鲤鱼）的卵母细胞中进行表征，由两个常见的成分Cdc2激酶和细胞周期蛋白B组成。尽管Xenopus和Goldfish卵母细胞中MPF的成分是同一两种物种的机制，但在这两种物种之间是不同的。在金鱼中，17Alpha，20beta-dp诱导不成熟的卵母细胞合成细胞周期蛋白B，进而通过其苏氨酸（THR161）磷酸化激活了35 kDa cdc2激酶，从而产生34 kDa活跃的CDC2 Kinase。这些在鱼类卵母细胞中MPF激活的机制显然与爪蟾不同，其中未成熟的卵母细胞中存在细胞周期蛋白B，并与Cdc2激酶（PREMPF）形成复合物。此外，在爪蟾卵母细胞中，Cdc2激酶激活需要苏氨酸（Thr14）和酪氨酸（Tyr15）的去磷酸化。 However, it is doubtful that tyrosine (Tyr15) dephosphorylation of cdc2 kinase is not a prerequisite for its activation during goldfish oocyte maturation, since the activation is not inhibited by vanadade, a protein phosphatase inhibitor commonly used for inhibiting cdc25 activity that dephosphorylates tyrosine (Tyr15) of cdc2 kinase, thereby inhibits its activation.较少的

项目成果

Iwao,Y.,Sakamoto,N.,Takahara,K.,Yamashita,M.and Nagahama,Y.: "The egg nucleus regulates the behavior of sperm nuclei as well as cycling of MPF in physiologically polyspermic necot eggs." Developmentol Biology. 160. 15-27 (1993)

Iwao,Y.、Sakamoto,N.、Takahara,K.、Yamashita,M. 和 Nagahama,Y.：“卵核调节精子核的行为以及生理多精新生卵中 MPF 的循环。”

Katsu,Y.,Yamashita,M.,Kajiura,H.and Nagahama,Y.(1993): "Behaviour of the components of maturaion-promoting factor,cdc2 kinase and cyclin B,during oocyte maturation of goldfish" Developmental Biology.

Katsu,Y.、Yamashita,M.、Kajiura,H. 和 Nagahama,Y.(1993)：“金鱼卵母细胞成熟过程中成熟促进因子、cdc2 激酶和细胞周期蛋白 B 成分的行为”发育生物学。

Natsuyama,S.,Noda,Y.,Yamashita,M.,Nagahama,Y.and Mori,T.: "Superoxide dismutase and thioiredoxin restore defective p34^<cdc2>Kinase activation in mouse two-cell block." Biochemica et Biophysica Acta. 1176. 90-94 (1993)

Natsuyama,S.、Noda,Y.、Yamashita,M.、Nagahama,Y. 和 Mori,T.：“超氧化物歧化酶和硫氧还蛋白恢复小鼠双细胞块中有缺陷的 p34^<cdc2> 激酶激活。”

Tokumoto,T.,Yamashita,M.,Yoshikuni,M.,and Nagahama,Y.: "Purification of latent proteasome(20S proteasome)and demonstration of active proteasome in goldfish(Carassuis anratus)oocyte cytosol" Journal of Biological Chemistry.

Tokumoto,T.、Yamashita,M.、Yoshikuni,M. 和 Nagahama,Y.：“金鱼（Carassuis anratus）卵母细胞胞质液中潜在蛋白酶体（20S 蛋白酶体）的纯化和活性蛋白酶体的演示”生物化学杂志。

Onoe,S.,Yamashita,M.,Kajiura,H.and Nagahama,Y.: "A fish homolog of the cdc2-related protein p40:its cDNA cloning and expression in oocytes" Biomedical Research. 14. 441-444 (1993)

Onoe,S.、Yamashita,M.、Kajiura,H. 和 Nagahama,Y.：“cdc2 相关蛋白 p40 的鱼类同源物：其 cDNA 克隆和在卵母细胞中的表达”生物医学研究。