Analysis of molecular regulation on female reproductive tract during fertilization using genetic model mice

遗传模型小鼠分析雌性生殖道受精过程的分子调控

• 批准号: 14571537
• 负责人: ARAKI Yoshihiko
• 金额: $ 2.11万
• 依托单位: Yamagata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571537/
• 关键词: female reproductive tract; fertilization; oviduct-specific glycoprotein; transgenic mouse; knock-out mouse; エストロゲン; プロモーター

The mammalian fertilization process takes place in a complex microenvironment within the female genital tract. A member of the chitinase protein family, oviduct-species glycoprotein (OGP), has been identified in oviductal fluid from various mammalian species including humans. Although it is thought that OGP is widely believed to involve in the process of mammalian fertilization, including sperm function and gamete interactions based on experimental results obtained in vitro, its physiological significance remains controversial. In this study, we generated transgenic mice using a 2.2-kilobase (kb) segment of the 5'-flanking sequence of the OGP gene for expression of the SV 40 T antigen (Tag). These mice spontaneously developed tumors in the female reproductive tract. Analysis using RT-PCR showed that the 2.2-kb OGP 5'-flanking region drove Tag mRNA expression in the oviduct, uterus, vagina and ovary, but not in other tissues. Ovariectomy suppressed Tag expression and thereby blocked tum … More origenesis in the transgenic mice. Estradiol administration to ovariectomized transgenic mice led to dramatic hyperplasia of the reproductive tract tissues in association with enhanced Tag expression, both in intensity and distribution. These results demonstrated that a 2.2-kb fragment of the 5'-flanking sequence of the mouse OGP gene was capable of directing the expression of Tag and inducing tumorigenesis in female reproductive tract tissues in an estrogen-dependent the expression of Tag and inducing tumorigenesis in female reproductive tract tissues in an estrogen-dependent manner. Estrogen response elements present in the promoter region were functional in vivo. In In addition, we eatablished OGP gene-null (ogp^<-/-> mice, and primarily characterized their reproductive properties to study the physiological function(s) of OGP. Data obtained from studies using in vivo or in vitro system showed that the fertility of ogp^<-/-> females was within normal limits. These results indicate that OGP is dispensable for the process of in vivo fertilization, at least in mice Less

哺乳动物的受精过程发生在女性生殖道内复杂的微环境中。输卵管糖蛋白（OGP）是几丁质酶蛋白家族的一员，已在包括人类在内的多种哺乳动物的输卵管液中鉴定出。尽管基于体外实验结果，OGP被广泛认为参与哺乳动物受精过程，包括精子功能和配子相互作用，但其生理意义仍存在争议。在这项研究中，我们使用OGP基因5 '侧翼序列的2.2-内切酶（kb）片段产生转基因小鼠，用于表达SV 40 T抗原（Tag）。这些小鼠自发地在雌性生殖道中产生肿瘤。RT-PCR分析表明，2.2 kb的OGP 5 '侧翼区驱动Tag mRNA在输卵管、子宫、阴道和卵巢中表达，但在其他组织中不表达。卵巢切除术抑制Tag表达，从而阻断肿瘤细胞的生长。 ...更多信息 转基因小鼠的起源。给予卵巢切除的转基因小鼠雌二醇导致生殖道组织的显著增生，与增强的Tag表达相关，无论是在强度还是分布上。这些结果表明，小鼠OGP基因的5 ′侧翼序列的2.2-kb片段能够指导Tag的表达并以雌激素依赖性方式诱导雌性生殖道组织中的肿瘤发生。雌激素反应元件存在于启动子区的功能在体内。此外，我们还建立了OGP基因敲除（ogp^-/->）小鼠模型，并对其生殖特性进行了初步研究，以探讨OGP的生理功能。从使用体内或体外系统的研究中获得的数据表明，ogp^<-/->雌性的生育力在正常范围内。这些结果表明，OGP在体内受精过程中是不稳定的，至少在小鼠中是如此。

项目成果

Araki et al.: "Immortalized epididymal cell lines from transgenic mice over expressing temperature-sensitive siman virus 40 T-antigen gene."J.Androl.. 23. 854-869 (2002)

Araki 等人：“转基因小鼠的永生化附睾细胞系过度表达温度敏感的西曼病毒 40 T 抗原基因。”J.Androl.. 23. 854-869 (2002)

Takeda et al.: "Expression of GPI-80, a β integrin-associated glycosylphosphatidyl inositol anchored protein, requires neutrophil differentiation with dimethylsulfoxide in HL-60 cells"Exp.Cell Res.. 286. 199-208 (2003)

Takeda 等人：“GPI-80（一种 β 整联蛋白相关糖基磷脂酰肌醇锚定蛋白）的表达需要在 HL-60 细胞中用二甲亚砜进行中性粒细胞分化”Exp.Cell Res. 286. 199-208 (2003)

Miyoshi et al.: "A mouse transgenic for murine oviduct-specific glycoprotein promoter-driven simian virus 40 large T-antigen : Tumor formation and its hormonal regulation."Mol.Reprod.Dev.. 63. 168-176 (2002)

Miyoshi 等人：“小鼠输卵管特异性糖蛋白启动子驱动的转基因猴病毒 40 大 T 抗原：肿瘤形成及其激素调节。”Mol.Reprod.Dev.. 63. 168-176 (2002)

Araki Y.: "Formation and structure of mammalian ovaries. In : Introduction to Mammalian Reproduction (Tulsiani DRP ed.) P141-153."Kluwer Academic Publishers, Norwell, MA, USA.. 403 (2003)

Araki Y.：“哺乳动物卵巢的形成和结构。见：哺乳动物生殖简介（Tulsiani DRP 版）P141-153。”Kluwer 学术出版社，诺威尔，马萨诸塞州，美国.. 403 (2003)

Takeda et al.: "Expression of GPI-80, a β2 integrin-associated glycosylphosphatidyl inositol anchored protein, requires neutrophil differentiation with dimethylsulfoxide in HL-60 cells."Exp.Cell Res.. 286. 199-208 (2003)

Takeda 等人：“GPI-80（一种 β2 整联蛋白相关糖基磷脂酰肌醇锚定蛋白）的表达需要在 HL-60 细胞中用二甲基亚砜进行中性粒细胞分化。”Exp.Cell Res. 286. 199-208 (2003)