Molecular basis of spermatogenesis and fertilization in mammals

哺乳动物精子发生和受精的分子基础

基本信息

  • 批准号:
    15208033
  • 负责人:
  • 金额:
    $ 27.96万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
  • 财政年份:
    2003
  • 资助国家:
    日本
  • 起止时间:
    2003 至 2005
  • 项目状态:
    已结题

项目摘要

To elucidate the molecular basis of mammalian spermatogenesis and fertilization, we have examined the synthesis, processing, and functions of sperm proteins involved in fertilization. The following experimental results have been obtained :1. In mouse, two different isoforms of ADAM1, ADAM1a and ADAM1b, are produced in the testis. ADAM1a is localized within the endoplasmic reticulum of testicular germ cells (TGC), whereas epididymal sperm contain only ADAM1b on the plasma membrane. We show that the loss of ADAM1a results in the male infertility because of the severely impaired ability of sperm to migrate from the uterus into the oviduct through the uterotubal junction. Among testis (sperm)-specific proteins examined, only the level of ADAM3 was strongly reduced in ADAM1a-deficient mouse sperm. Moreover, the appearance of ADAM3 on the sperm surface was dependent on the formation of a fertilin protein complex between ADAM1a and ADAM2 in TGC. These results suggest that ADAM1a/ADAM2 fertili … More n may be implicated in the selective transport of specific sperm proteins including ADAM3 from the endoplasmic reticulum of testicular germ cells onto the cell surface.We also show that mutant male mice lacking ADAM1b are fertile, and the loss of ADAM1b results in no significant defect in the sperm functions. ADAM1b-deficient epididymal sperm showed a severe reduction of ADAM2 on the cell surface, despite the normal presence of ADAM2 in TGC. The appearance of ADAM1b and ADAM2 on the sperm surface depended on formation and abundance of ADAM1b/ADAM2 fertilin in TGC. Thus, mouse ADAM1b/ADAM2 fertilin may play the crucial role not in the sperm/egg fusion, but in the appearance of these two ADAMs on the sperm surface.2. A glycosylphosphatidylinositol (GPI)-anchored hyaluronidase, PH-20, on the sperm surface has long been believed to assist sperm penetration through the cumulus mass surrounding the eggs. However, mouse sperm lacking PH-20 were still capable of penetrating the cumulus mass despite a delayed dispersal of cumulus cells. Intriguingly, a 55-kDa hyaluronan-hydrolyzing protein was abundantly present in wild-type and PH-20-deficient mouse sperm. In this study, we have purified the 55-kDa mouse protein from soluble protein extracts released from epididymal sperm by acrosome reaction, and have identified as a novel hyaluronidase, Hyal5. Hyal5 was exclusively expressed in the testis, and formed a 160-kbp gene cluster together with Hyalp1, Hyal4, and Ph-20 on mouse chromosome 6. Hyal5 was a single-chain hyaluronidase present on the plasma and acrosomal membranes of sperm presumably as a GPI-anchored protein. Moreover, hyaluronan zymography revealed that Hyal5 is enzymatically active in the pH range 5-7, and inactive at pH 3 and 4. Both Hyal5-enriched, PH-20-free soluble protein extracts, and PH-20-deficient mouse sperm were capable of dispersing cumulus cells from the cumulus mass. The cumulus cell dispersal was strongly inhibited by the presence of a hyaluronidase inhibitor, apigenin. These results suggest that in the mouse, Hyal5 may function principally as a "cumulus matrix depolymerase" in the sperm penetration through the cumulus mass, and in the local hyaluronan hydrolysis near or on the surface of the egg zona pellucida to enable the proximal region of sperm tail to move freely. PH-20 may compensate in part for the functional roles of Hyal5.3. We have previously identified a 42-kDa serine protease, TESP5, identical to testisin, esp-1, or tryptase 4 as a candidate enzyme involved in the sperm penetration of egg ZP. Mouse TESP5 is glycosylphosphatidylinositol (GPI)-anchored on the cell surface of cauda epididymal sperm. To elucidate the role(s) of TESP5 in fertilization, we have produced mutant mice carrying a targeted mutation in the TESP5 gene by homologous recombination in embryonic stem cells. The TESP5-deficient male and female mice showed a normal fertility. However, in vitro fertilization assay revealed that epididymal sperm lacking TESP5 barely fertilize metaphase II-arrested oocytes. The loss of TESP5 on sperm resulted in the reduced ability to bind the ZP in vitro. Even when TESP5-deficient sperm bound to the ZP, the rate of ZP-induced acrosome reaction was very low. Moreover, the ability of sperm to fuse with egg in vitro was severely impaired by the TESP5 loss. These results suggest that TESP5 may act in the acrosome reaction, and imply that the impaired function of TESP5-deficient sperm may be compensated for during the transit through the uterus and/or oviduct. Less
为了阐明哺乳动物精子发生和受精的分子基础,我们研究了精子蛋白的合成、加工和功能。取得了如下实验结果:1.在小鼠中,在睾丸中产生两种不同的ADAM 1亚型,ADAM 1a和ADAM 1b。ADAM 1a位于睾丸生殖细胞(TGC)的内质网内,而附睾精子仅在质膜上含有ADAM 1b。我们发现,ADAM 1a的缺失导致男性不育,因为精子从子宫通过子宫输卵管连接处迁移到输卵管的能力严重受损。在检测的睾丸(精子)特异性蛋白质中,只有ADAM 1a缺陷小鼠精子中的ADAM 3水平显著降低。此外,在TGC中,精子表面的ADAM 3的出现依赖于ADAM 1a和ADAM 2之间的受精素蛋白复合物的形成。这些结果表明,ADAM 1a/ADAM 2可育, ...更多信息 n可能参与特异性精子蛋白包括ADAM 3从睾丸生殖细胞内质网到细胞表面的选择性转运,我们还表明,缺乏ADAM 1b的突变雄性小鼠是有生育能力的,并且ADAM 1b的缺失不会导致精子功能的显著缺陷。ADAM 1b缺陷的附睾精子表现出细胞表面的ADAM 2严重减少,尽管TGC中的ADAM 2正常存在。TGC中精子表面的ADAM 1b和ADAM 2的出现依赖于ADAM 1b/ADAM 2受精素的形成和丰度。因此,小鼠ADAM 1b/ADAM 2受精蛋白可能不是在精卵融合中起关键作用,而是在精子表面这两种亚当斯的出现中起关键作用.长期以来,人们一直认为精子表面的糖基磷脂酰肌醇(GPI)锚定透明质酸酶PH-20有助于精子穿透卵子周围的卵丘。然而,缺乏PH-20的小鼠精子仍然能够穿透卵丘质量,尽管卵丘细胞的分散延迟。有趣的是,一个55 kDa的透明质酸水解蛋白大量存在于野生型和PH-20缺陷小鼠精子。在这项研究中,我们已经纯化了55 kDa的小鼠蛋白质从附睾精子顶体反应释放的可溶性蛋白提取物,并已确定为一种新的透明质酸酶,Hyal 5。Hyal 5仅在睾丸中表达,并与Hyalp 1、Hyal 4和Ph-20一起在小鼠6号染色体上形成160-kbp的基因簇。Hyal 5是存在于精子的质膜和顶体膜上的单链透明质酸酶,推测为GPI锚定蛋白。此外,透明质酸酶谱显示Hyal 5在pH 5-7范围内具有酶活性,而在pH 3和4下无活性。Hyal 5富集,PH-20自由可溶性蛋白提取物,PH-20缺乏小鼠精子能够分散卵丘细胞从卵丘质量。的卵丘细胞分散强烈抑制透明质酸酶抑制剂,芹菜素的存在。这些结果表明,在小鼠中,Hyal 5可能主要作为“卵丘基质解聚酶”在精子穿透卵丘团中起作用,并且在卵透明质膜表面附近或表面上的局部透明质酸水解中起作用,以使精子尾部的近端区域自由移动。PH-20可以部分补偿Hyal5.3的功能作用。我们以前已经确定了一个42 kDa的丝氨酸蛋白酶,TESP 5,相同的testisin,ESP-1,或类胰蛋白酶4作为候选酶参与精子穿透卵ZP。小鼠TESP 5是锚定在附睾尾部精子细胞表面的糖基磷脂酰肌醇(GPI)。为了阐明TESP 5在受精中的作用,我们通过在胚胎干细胞中同源重组产生了携带TESP 5基因靶向突变的突变小鼠。TESP 5缺陷的雄性和雌性小鼠显示出正常的生育能力。然而,体外受精试验表明,缺乏TESP 5的附睾精子几乎不能使中期II停滞的卵母细胞受精。精子上TESP 5的缺失导致体外结合ZP的能力降低。即使TESP 5缺陷精子与ZP结合,ZP诱导的顶体反应率也非常低。此外,TESP 5的缺失严重损害了精子与卵子在体外融合的能力。这些结果表明,TESP 5可能在顶体反应中起作用,并暗示TESP 5缺陷精子的受损功能可能在通过子宫和/或输卵管的过程中得到补偿。少

项目成果

期刊论文数量(62)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Zhuang Tiangang: "Transgenic expression of testis-specific poly(A) polymerase TPAP in wild-type and TPAP-deficient mice"Journal of Reproduction and Development. (in press). (2004)
庄天罡:“睾丸特异性多聚A聚合酶TPAP在野生型和TPAP缺陷小鼠中的转基因表达”生殖与发育杂志。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Fujita Toshihide: "Full activation of estrogen receptor α activation function-1 induces proliferation of breast cancer cells"Journal of Biological Chemistry. 278. 26704-26714 (2003)
Fujita Toshihide:“雌激素受体α激活功能-1的完全激活诱导乳腺癌细胞增殖”《生物化学杂志》278. 26704-26714(2003)。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Transgenic expression of testis-specific poly(A) polymerase TPAP in wild-type and TPAP-deficient mice
Synthesis, processing, and subcellular localization of mouse ADAM3 during spermatogenesis and epididymal sperm transport
Possible function of the ADAM1a/ADAM2 fertilin complex in the appearance of ADAM3 on the sperm surface
  • DOI:
    10.1074/jbc.m314249200
  • 发表时间:
    2004-08-13
  • 期刊:
  • 影响因子:
    4.8
  • 作者:
    Nishimura, H;Kim, E;Baba, T
  • 通讯作者:
    Baba, T
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

BABA Tadashi其他文献

BABA Tadashi的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('BABA Tadashi', 18)}}的其他基金

Functional Regulation of Migrating Sperm in the Mouse Oviduct
小鼠输卵管中精子迁移的功能调节
  • 批准号:
    18K06241
  • 财政年份:
    2018
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Basic and Applied Studies on Regulation of Gene Expression by Small DNAs
小DNA调控基因表达的基础与应用研究
  • 批准号:
    26650004
  • 财政年份:
    2014
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Integrated analyses of virulence factors from Staphylococcus aureusaiming to pursue novel pathogenic mechanisms and chemotherapy based on new concept
金黄色葡萄球菌毒力因子综合分析,寻求新的致病机制及新理念化疗
  • 批准号:
    22590402
  • 财政年份:
    2010
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Basic Studies on Mammalian Fertilization and Implantation
哺乳动物受精和着床的基础研究
  • 批准号:
    21248038
  • 财政年份:
    2009
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Regulatory Mechanism of Sperm Function in Female Reproductive Tract
女性生殖道精子功能的调节机制
  • 批准号:
    18208029
  • 财政年份:
    2006
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Studies on the Regulation of Fertilization and Activation of Development
受精调控与发育激活研究
  • 批准号:
    12306018
  • 财政年份:
    2000
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Molecular Mechanism of Functional Regulation of Spermatiogenic Cells by Interaction with Other Somatic Cells and Eggs
生精细胞与其他体细胞和卵子相互作用功能调节的分子机制
  • 批准号:
    11235201
  • 财政年份:
    1999
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas
Functional alteration of cereal starches using the genes encoding starch-synthesizing enzymes
使用编码淀粉合成酶的基因改变谷物淀粉的功能
  • 批准号:
    10556029
  • 财政年份:
    1998
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Studies on Sperm Function in Fertilization
精子在受精过程中的功能研究
  • 批准号:
    09460154
  • 财政年份:
    1997
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Studies on Molecular Mechanism of Starch Biosynthesis
淀粉生物合成的分子机制研究
  • 批准号:
    07660092
  • 财政年份:
    1995
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)

相似国自然基金

茄子果形调控基因EGG的克隆和机制解析
  • 批准号:
    32302584
  • 批准年份:
    2023
  • 资助金额:
    30 万元
  • 项目类别:
    青年科学基金项目
双靶向EgG1Y162重组多表位海藻酸钠-壳聚糖纳米缓释疫苗的制备及免疫机制研究
  • 批准号:
    32260192
  • 批准年份:
    2022
  • 资助金额:
    33 万元
  • 项目类别:
    地区科学基金项目
双靶向DC和M细胞的EgA31-EgG1Y162壳聚糖载体递送系统疫苗的构建和免疫机制研究
  • 批准号:
    32160182
  • 批准年份:
    2021
  • 资助金额:
    35 万元
  • 项目类别:
    地区科学基金项目
基于海藻酸“Egg-box”自组装结构的锂离子混合电容器
  • 批准号:
    51772187
  • 批准年份:
    2017
  • 资助金额:
    60.0 万元
  • 项目类别:
    面上项目
细粒棘球绦虫EgA31-EgG1y162多价优势表位基因疫苗的筛选和保护机制的研究
  • 批准号:
    31560262
  • 批准年份:
    2015
  • 资助金额:
    38.0 万元
  • 项目类别:
    地区科学基金项目
细粒棘球绦虫BCG-egG1Y162重组疫苗的构建和保护机制研究
  • 批准号:
    31000411
  • 批准年份:
    2010
  • 资助金额:
    19.0 万元
  • 项目类别:
    青年科学基金项目

相似海外基金

Mammalian Fertilization: Identifying the second sperm factor that induces residual calcium oscillations and its contributions to egg activation
哺乳动物受精:识别诱导残留钙振荡的第二个精子因子及其对卵子激活的贡献
  • 批准号:
    10574938
  • 财政年份:
    2022
  • 资助金额:
    $ 27.96万
  • 项目类别:
Identifying sperm-egg receptor pairs essential for mammalian fertilization to select new targets for fertility treatment and contraception.
识别哺乳动物受精所必需的精卵受体对,以选择生育治疗和避孕的新靶标。
  • 批准号:
    MR/M012468/1
  • 财政年份:
    2015
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Research Grant
Molecular mechanism of egg-sperm interaction during fertilization in teleost and development of application system for artificial fertilization
硬骨鱼受精过程中卵子与精子相互作用的分子机制及人工受精应用系统的开发
  • 批准号:
    23380110
  • 财政年份:
    2011
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular Mechanism of sperm-egg membrane fusion in fertilization
受精过程中精卵膜融合的分子机制
  • 批准号:
    16390480
  • 财政年份:
    2004
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Dissertation Research: Dynamics of Fertilization: Ecological Consequences of Flow on Sperm-egg Interactions
论文研究:受精动力学:流动对精子-卵子相互作用的生态影响
  • 批准号:
    0206775
  • 财政年份:
    2002
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Standard Grant
Identification of sperm-derived egg-activating protein at fertilization of mammalian eggs
哺乳动物卵子受精时精子来源的卵子激活蛋白的鉴定
  • 批准号:
    13470010
  • 财政年份:
    2001
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
BIOCHEMISTRY OF SPERM-EGG INTERACTION AND FERTILIZATION
精卵相互作用和受精的生物化学
  • 批准号:
    2853654
  • 财政年份:
    1999
  • 资助金额:
    $ 27.96万
  • 项目类别:
STUDIES ON SPERM-EGG INTERACTIONSIN MAMMALIAN FERTILIZATION
哺乳动物受精过程中精卵相互作用的研究
  • 批准号:
    09044240
  • 财政年份:
    1997
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Identification of sperm-derived egg-activating factor and analysis of molecular mechanisms of intracellular Ca^<2+> increase at fertilization of mammalian eggs
哺乳动物卵子受精时精子源性卵子激活因子的鉴定及细胞内Ca^2>增加的分子机制分析
  • 批准号:
    08457016
  • 财政年份:
    1996
  • 资助金额:
    $ 27.96万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
SPERM GALACTOSYLTRANSFERASE AND EGG ZP3 IN FERTILIZATION
受精过程中的精子半乳糖基转移酶和卵子 ZP3
  • 批准号:
    3048997
  • 财政年份:
    1991
  • 资助金额:
    $ 27.96万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了