Minimum Y gene complement necessary for successful ART

成功 ART 所需的最低 Y 基因补充

• 批准号: 7863953
• 负责人: Monika A Ward
• 金额: $ 0.67万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2009-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7863953
• 关键词: Acrosome; Age; Assisted Reproductive Technology; Bypass; Cells; Chromosomes; Code; Competence; Complement; Complement component C1; Data; Development; Embryo; Embryo Transfer; Embryonic Development; Event; Fertilization; Gene Deletion; Gene Targeting; Generations; Genes; Genetic Recombination; Genotype; Germ Cells; Goals; Head; Homologous Gene; Human; Infertility; Injection of therapeutic agent; Intracytoplasmic Sperm Injections; Life; Male Infertility; Maps; Meiosis; Modeling; Mus; Oocytes; Partner in relationship; Phenotype; Process; Production; Reproduction; Research; Role; Spermatids; Spermatogenesis; Spermatogenic Cell; Spermiogenesis; Staging; Testing; Testis; Transgenes; Upper arm; Work; Y Chromosome; assisted reproduction; cell type; deletion analysis; fusion gene; gene function; in vivo; interest; male; mouse model; offspring; positional cloning; public health relevance; research study; sperm cell; sperm function; sry Genes; zygote

DESCRIPTION (provided by applicant): It has been argued that most Y chromosome coded genes are likely to have roles in sperm production or function. However, it is not clear if these genes provide essential spermatogenic function or just potentiate the spermatogenic process. The goal of this application is to determine the minimum Y chromosome gene requirement that is compatible with successful reproduction by assisted reproductive technologies (ART). The hypothesis of this application is that a Y gene complement of as few as two or three genes is enough to enable the production of male `gametes' (round spermatids or sperm) that are capable of participating in fertilization if delivered into the oocytes via injection. In preliminary data we provide evidence that in the mouse the presence of only two Y-coded genes, Sry and Eif2s3y, allows formation of testes, ongoing spermatogonial proliferation, and completion of meiosis to generate round spermatids. We suggest that further addition of one copy of Zfy is sufficient to allow the production of some sperm, albeit with morphologically abnormal heads. We also show that assisted reproduction by ICSI and IVF enable the generation of live offspring from subfertile and infertile males with Y chromosome deficiencies. In this application we will focus on analyzing various mouse models with limited Y gene complements: (1) males with an almost intact Y short arm but complete absence of Y long arm genes; (2) males with no Y long arm genes and the known Y short arm genes limited to Sry, Eif2s3y, a single copy of Zfy, and a reduced number of copies of Rbmy; and (3) males with only Eif2s3y and Sry. In Specific Aim 1 we will generate these males with limited Y gene complements and define more precisely at what stage of spermiogenesis cells arrest or become abnormal. We will perform histological analysis of the testes, confirm the presence of specific spermatogenic cell types by immunostaining, and examine acrosome development as a marker of spermiogenic stage. In Specific Aim 2 we will use sperm and/or round spermatids from these models for ICSI and/or ROSI. We will observe early post-fertilization events after injection, obtain embryos, and produce live offspring. We will genotype progeny to test which sperm genotypes were successful in supporting fertilization and embryo development. The significance of this application is that it will advance the understanding of the role of Y chromosome encoded genes in spermatogenesis and sperm function; it will also provide valuable information for those using ART to treat human infertility associated with Y gene deficiencies. PUBLIC HEALTH RELEVANCE: In this application we seek to determine the minimum Y gene complement that is compatible with the generation of sperm competent in fertilization via ART (ICSI and ROSI). The study will add to the understanding of the functions of Y chromosome coded genes by defining which Y genes provide essential spermatogenic function rather than just potentiating the spermatogenic process. The application also has significance for those utilizing ART to treat male infertility.

描述（由申请人提供）：有人认为大多数 Y 染色体编码基因可能在精子的产生或功能中发挥作用。然而，尚不清楚这些基因是否提供必要的生精功能或只是增强生精过程。此应用的目标是确定与辅助生殖技术 (ART) 成功繁殖相兼容的最低 Y 染色体基因要求。该应用的假设是，只要两个或三个基因的 Y 基因互补就足以产生雄性“配子”（圆形精子细胞或精子），如果通过注射将其输送到卵母细胞中，则能够参与受精。在初步数据中，我们提供的证据表明，在小鼠中，仅存在两个 Y 编码基因 Sry 和 Eif2s3y，就可以形成睾丸、持续精原细胞增殖以及完成减数分裂以产生圆形精子细胞。我们建议进一步添加一份 Zfy 副本足以产生一些精子，尽管其头部形态异常。我们还表明，通过 ICSI 和 IVF 辅助生殖可以使 Y 染色体缺陷的生育力低下和不育男性产生活的后代。在本申请中，我们将重点分析具有有限Y基因互补的各种小鼠模型：（1）具有几乎完整的Y短臂但完全缺乏Y长臂基因的雄性； (2)无Y长臂基因的雄性，已知的Y短臂基因仅限于Sry、Eif2s3y、单个拷贝的Zfy和减少数量的Rbmy拷贝； (3) 仅具有 Eif2s3y 和 Sry 的雄性。在具体目标 1 中，我们将生成这些具有有限 Y 基因互补体的雄性，并更精确地定义精子发生细胞在哪个阶段停滞或变得异常。我们将对睾丸进行组织学分析，通过免疫染色确认特定生精细胞类型的存在，并检查顶体发育作为生精阶段的标志。在具体目标 2 中，我们将使用这些模型中的精子和/或圆形精子细胞进行 ICSI 和/或 ROSI。我们将观察注射后早期受精后事件，获得胚胎，并产生活的后代。我们将对后代进行基因分型，以测试哪些精子基因型成功支持受精和胚胎发育。这一应用的意义在于，它将推进对Y染色体编码基因在精子发生和精子功能中作用的理解；它还将为那些使用 ART 治疗与 Y 基因缺陷相关的人类不孕症的人们提供有价值的信息。公共健康相关性：在本申请中，我们寻求确定与通过 ART（ICSI 和 ROSI）产生具有受精能力的精子相容的最小 Y 基因补充。该研究将通过定义哪些 Y 基因提供重要的生精功能而不仅仅是增强生精过程，来加深对 Y 染色体编码基因功能的理解。该应用对于利用 ART 治疗男性不育症也具有重要意义。

项目成果

Live offspring from mice lacking the Y chromosome long arm gene complement.

缺乏 Y 染色体长臂基因补体的小鼠的活后代。

• 发表时间: 2009-08
• 影响因子: 3.6
• 作者: Yamauchi, Yasuhiro;Riel, Jonathan M;Wong, Samantha J;Ojarikre, Obah A;Burgoyne, Paul S;Ward, Monika A
• 通讯作者: Ward, Monika A

The multicopy gene Sly represses the sex chromosomes in the male mouse germline after meiosis.

多拷贝基因 Sly 在减数分裂后抑制雄性小鼠种系中的性染色体。

• 发表时间: 2009-11
• 影响因子: 9.8
• 作者: Cocquet, Julie;Ellis, Peter J I;Yamauchi, Yasuhiro;Mahadevaiah, Shantha K;Affara, Nabeel A;Ward, Monika A;Burgoyne, Paul S
• 通讯作者: Burgoyne, Paul S

Deficiency in the multicopy Sycp3-like X-linked genes Slx and Slxl1 causes major defects in spermatid differentiation.

多拷贝 Sycp3 样 X 连锁基因 Slx 和 Slxl1 的缺陷会导致精子细胞分化的重大缺陷。

• 发表时间: 2010-10-15
• 影响因子: 3.3
• 作者: Cocquet, Julie;Ellis, Peter J I;Yamauchi, Yasuhiro;Riel, Jonathan M;Karacs, Thomas P S;Rattigan, Aine;Ojarikre, Obah A;Affara, Nabeel A;Ward, Monika A;Burgoyne, Paul S
• 通讯作者: Burgoyne, Paul S

Deficiency in mouse Y chromosome long arm gene complement is associated with sperm DNA damage.

小鼠 Y 染色体长臂基因补体的缺陷与精子 DNA 损伤有关。

• 发表时间: 2010
• 影响因子: 12.3
• 作者: Yamauchi, Yasuhiro;Riel, Jonathan M;Stoytcheva, Zoia;Burgoyne, Paul S;Ward, Monika A
• 通讯作者: Ward, Monika A

Blastomere removal from cleavage-stage mouse embryos alters steroid metabolism during pregnancy.

从卵裂期小鼠胚胎中去除卵裂球会改变怀孕期间的类固醇代谢。

• 发表时间: 2012-07
• 影响因子: 3.6
• 作者: Sugawara, Atsushi;Sato, Brittany;Bal, Elise;Collier, Abby C;Ward, Monika A
• 通讯作者: Ward, Monika A