Do we need Y chromosome for successful reproduction?

我们需要Y染色体才能成功繁殖吗？

• 批准号: 10377939
• 负责人: Monika A Ward
• 金额: $ 44.29万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-25 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377939
• 关键词: Address; Adult; Asexual Reproduction; Biological Models; Birth; Breeding; Cell Line; Cell Transplantation; Cells; Chimera organism; Chromosomes; Development; Ear; Embryo; Engineering; Female; Fertility; Fertilization; Fetal Development; Fibroblasts; Gametogenesis; Gender; Generations; Genes; Genetic Engineering; Germ Cells; Haploidy; Homologous Gene; In Vitro; Individual; Infertility; Laboratories; Mammals; Meiotic Recombination; Modeling; Mus; National Institute of Child Health and Human Development; Neonatal; Oocytes; Oogenesis; Ovarian; Ovary; Paper; Production; Publishing; Reproduction; Sex Chromosomes; Sex Differences; Somatic Cell; Spermatogenesis; Structure of primordial sex cell; Technology; Testing; Testis; Transgenic Mice; Transgenic Organisms; Transplantation; X Chromosome; Y Chromosome; assisted reproduction; fertility preservation; fetal; gene complementation; imprint; induced pluripotent stem cell; innovative technologies; interest; male; mouse model; offspring; overexpression; preservation; reconstitution; reconstruction; sex; sex determination; sperm cell; stem cell differentiation; symposium

Mammalian reproduction conventionally requires oocytes provided by a female and sperm provided by a male to achieve fertilization. We go beyond this convention and propose the hypothesis that an adult individual of any sex can be induced to produce gametes of the opposite sex, and that these gametes are functional in assisted fertilization. The premise for this hypothesis comes from our recently published studies demonstrating that genetically engineered male mice with limited or no Y chromosome genes can successfully reproduce by assisted fertilization (ART), and from significant advancements in field of cell reprogramming and differentiation. Our laboratory has shown that in the mouse only two Y chromosome genes, testis determinant Sry and spermatogenesis driver Eif2s3y, are sufficient for a male to produce haploid male gametes functional in ART. We have subsequently demonstrated that the function of these two genes could be replaced by that of their homologues encoded on other chromosomes, and that a mouse with a single X chromosome (XO) lacking all Y chromosome genes can produce male gametes and sire healthy offspring after assisted fertilization. The laboratory of our collaborator, Mitinori Saitou, has shown that both male and female gametes can be obtained from induced pluripotent stem cells (iPSC) differentiated into primordial germ cell like-cells (PGCLC). In this proposal, we marry our findings and expertise and ask 3 questions: Aim 1. Can an adult female mouse produce male gametes functional in assisted fertilization? Aim 2. Can an adult male mouse produce female gametes functional in assisted fertilization? Aim 3. Can an adult mouse of either sex sire uniparental offspring? To address these questions we will develop somatic cell lines from an adult mouse of a given sex, reprogram to iPSC, identify the clones that have lost one sex chromosome and became XO, and differentiate into PGCLC. To produce male gametes we will transgenically add a spermatogenesis driver and transplant PGCLC to testes from neonatal males. To produce female gametes we will reconstitute ovaries in vitro and transplant them under ovarian bursa of recipient females. We will test the function of such derived male and female gametes using assisted reproduction. To produce uniparental offspring, male and female gametes derived from the same individual will be used for fertilization. The findings from this project will impact on our understanding of sex specific differences, especially pertaining to effects of sex chromosomes and X and Y genes on germline development. If successful, we will also provide the field with a proof-of-principle that offspring of both sexes can be obtained from a single individual, which will impact on species preservation.

哺乳动物的繁殖通常需要雌性提供的卵母细胞和雄性提供的精子才能实现受精。我们超越了这一传统，提出了这样的假设，即任何性别的成年个体都可以被诱导产生异性配子，这些配子在辅助受精中起作用。这一假设的前提来自我们最近发表的研究，该研究表明，具有有限Y染色体基因或没有Y染色体基因的基因工程雄性小鼠可以通过辅助受精(ART)成功繁殖，以及细胞重新编程和分化领域的重大进步。我们的实验室已经证明，在小鼠中，只有两个Y染色体基因，即睾丸决定基因Sry和精子发生驱动基因Eif2s3y，足以让雄性产生在ART中具有功能的单倍体雄性配子。我们随后证明，这两个基因的功能可以被它们在其他染色体上编码的同源基因所取代，并且具有单个X染色体(XO)的小鼠在辅助受精后可以产生雄配子并产生健康的后代。我们的合作者Mitinori Saitou的实验室已经证明，通过诱导多能干细胞(IPSC)分化为原始生殖细胞样细胞(PGCLC)，可以获得雄配子和雌配子。在这项提案中，我们结合我们的发现和专业知识，提出三个问题：目的1.成年雌性小鼠能否产生在辅助受精中起作用的雄配子？目的2.成年雄性小鼠能否产生具有辅助受精功能的雌配子？目的3.成年小鼠，无论性别，都可以单亲繁殖后代吗？为了解决这些问题，我们将从一只给定性别的成年小鼠身上培养出体细胞系，重新编程到iPSC，鉴定失去一条性染色体并成为XO的克隆，并分化为PGCLC。为了产生雄配子，我们将转基因添加一个精子发生驱动器，并将PGCLC移植到新生儿雄性的睾丸中。为了产生雌配子，我们将在体外重建卵巢，并将它们移植到受体雌性的卵巢囊下。我们将使用辅助生殖来测试这种衍生的雄配子和雌配子的功能。为了产生单亲后代，来自同一个体的雄配子和雌配子将用于受精。这个项目的发现将影响我们对性别特异性差异的理解，特别是关于性染色体以及X和Y基因对生殖系发育的影响。如果成功，我们还将向该领域提供一项原则证明，即两性后代可以从单一个体获得，这将影响物种保护。

项目成果

Macroscopic demonstration of the male urogenital system with evidence of a direct inguinal hernia utilizing room temperature plastination.

利用室温塑化技术对男性泌尿生殖系统进行宏观展示，并提供直接腹股沟疝的证据。

• DOI: 10.2399/ana.16.036
• 发表时间: 2016
• 期刊: Anatomy
• 作者: Ruthig,VictorA;Labrash,Steven;Lozanoff,Scott;Ward,MonikaA
• 通讯作者: Ward,MonikaA

Genetic variation in the Y chromosome and sex-biased DNA methylation in somatic cells in the mouse.

• DOI: 10.1007/s00335-022-09970-z
• 发表时间: 2023-03
• 期刊: MAMMALIAN GENOME
• 影响因子: 2.5
• 作者: Batdorj, Enkhjin;AlOgayil, Najla;Zhuang, Qinwei Kim-Wee;Galvez, Jose Hector;Bauermeister, Klara;Nagata, Kei;Kimura, Tohru;Ward, Monika A.;Taketo, Teruko;Bourque, Guillaume;Naumova, Anna K.
• 通讯作者: Naumova, Anna K.

Oocytes from female mice on MF1 genetic background are not suitable for assisted reproduction†.

来自具有 MF1 遗传背景的雌性小鼠的卵母细胞不适合辅助生殖。

• DOI: 10.1093/biolre/ioz224
• 发表时间: 2020
• 期刊: Biology of reproduction
• 影响因子: 3.6
• 作者: Yamauchi,Yasushiro;Ajduk,Anna;Ward,MonikaA
• 通讯作者: Ward,MonikaA

Rescue of Sly Expression Is Not Sufficient to Rescue Spermiogenic Phenotype of Mice with Deletions of Y Chromosome Long Arm.

拯救 Sly 表达不足以拯救 Y 染色体长臂缺失小鼠的生精表型。

• DOI: 10.3390/genes10020133
• 发表时间: 2019
• 期刊: Genes
• 影响因子: 3.5
• 作者: Riel,JonathanM;Yamauchi,Yasuhiro;Ruthig,VictorA;Malinta,QushayU;Blanco,Mélina;Moretti,Charlotte;Cocquet,Julie;Ward,MonikaA
• 通讯作者: Ward,MonikaA

Testicular abnormalities in mice with Y chromosome deficiencies.

Y染色体缺陷小鼠的睾丸异常。

• DOI: 10.1095/biolreprod.116.144006
• 发表时间: 2017
• 期刊: Biology of reproduction
• 影响因子: 3.6
• 作者: Ruthig,VictorA;Nielsen,Torbjoern;Riel,JonathanM;Yamauchi,Yasuhiro;Ortega,EgleA;Salvador,Quinci;Ward,MonikaA
• 通讯作者: Ward,MonikaA