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Do we need Y chromosome for successful reproduction?

我们需要Y染色体才能成功繁殖吗？

基本信息

批准号：
9915948
负责人：
Monika A Ward
金额：
$ 45.52万
依托单位：
UNIVERSITY OF HAWAII AT MANOA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-25 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9915948
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

项目摘要

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来实现受精。我们超越了这一惯例，提出了这样的假设：任何性别的成年个体都可以被诱导产生异性的配子，并且这些配子在辅助受精中发挥作用。这一假设的前提来自我们最近发表的研究，该研究表明，Y染色体基因有限或没有的基因工程雄性小鼠可以通过辅助受精（ART）成功繁殖，并且来自细胞重编程和分化领域的重大进展。我们的实验室表明，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 中发挥功能的单倍体雄配子。我们随后证明，这两个基因的功能可以被其他染色体上编码的同源基因的功能所取代，并且具有缺乏所有 Y 染色体基因的单个 X 染色体（XO）的小鼠可以在辅助受精后产生雄性配子并产生健康的后代。我们的合作者 Mitinori Saitou 的实验室表明，雄性和雌性配子都可以从分化为原始生殖细胞样细胞 (PGCLC) 的诱导多能干细胞 (iPSC) 中获得。在此提案中，我们将我们的发现和专业知识结合起来，并提出 3 个问题：目标 1. 成年雌性小鼠能否产生具有辅助受精功能的雄性配子？目标 2. 成年雄性小鼠能否产生具有辅助受精功能的雌性配子？目标 3. 任何性别的成年小鼠都能生出单亲后代吗？为了解决这些问题，我们将从特定性别的成年小鼠中开发体细胞系，重新编程为 iPSC，识别丢失一条性染色体并变成 XO 的克隆，并分化为 PGCLC。为了产生雄性配子，我们将通过转基因方式添加精子发生驱动因素，并将 PGCLC 移植到新生雄性的睾丸中。为了产生雌性配子，我们将在体外重建卵巢并将其移植到受体雌性的卵巢囊下。我们将使用辅助生殖测试此类衍生的雄性和雌性配子的功能。为了产生单亲后代，来自同一个体的雄性和雌性配子将用于受精。该项目的研究结果将影响我们对性别特异性差异的理解，特别是有关性染色体以及 X 和 Y 基因对种系发育的影响的理解。如果成功，我们还将为该领域提供一个原理证明，即可以从单个个体中获得两性后代，这将对物种保护产生影响。