Mechanisms of Sex Determination in Zebrafish

斑马鱼性别决定机制

• 批准号: 9230392
• 负责人: JOHN H. POSTLETHWAIT
• 金额: $ 27.08万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9230392
• 关键词: Address; Adolescent; Affect; Aromatase; Bacterial Artificial Chromosomes; Biological; Cell Communication; Cell Proliferation; Cessation of life; DNA Double Strand Break; Detection; Developed Countries; Developing Countries; Development; Disease; Elements; Environment; Environmental Risk Factor; Enzymes; Equilibrium; Estrogens; FOXL2 gene; Female; Fishes; Gene Expression; Genes; Genetic; Genotype; Germ Cells; Gonadal structure; Growth; Hand; Human; India; Individual; Induced Mutation; Investigation; Learning; Mammals; Meiosis; Methods; Modeling; Molecular Genetics; Mutation; Nature; Oocytes; Organ; Orthologous Gene; Outcome; Ovary; Pathway interactions; Polycystic Ovary Syndrome; Population; Pregnancy; Production; Reproductive Health; Resources; Role; Sex Bias; Signal Transduction; Signaling Molecule; Specific qualifier value; Structure of primordial sex cell; Suggestion; Supporting Cell; Syndrome; System; Temperature; Testicular Dysgenesis Syndrome; Testing; Testis; Testosterone; Transgenic Organisms; Uterus; Vertebrates; Woman; Work; Y Chromosome; Zebrafish; experimental study; extracellular; gene function; gene repair; genetic sex determination; in utero; innovation; insight; male; men; mutant; neuronal cell body; novel; nutrition; public health relevance; reference genome; reproductive; sex; sex determination; sry Genes; tool; transcriptome; transcriptomics; whole genome

DESCRIPTION (provided by applicant) Normal human gonad development can be adversely affected in utero by unknown genetic and environmental factors leading to testis dysgeneration syndrome and polycystic ovarian syndrome. In mammals, the Y-chromosome gene SRY initiates an incompletely understood genetic network that directs the gonad to become a testis, while competing genes direct ovary development. Non-mammalian vertebrates use similar down-stream networks, but primary sex determination (SD) genes differ. Additionally, in some species, environmental factors such as temperature or nutrition bias SD. Our broad objective is to learn how genetic factors interact with the environment to tip SD balance. Work focuses on zebrafish, a genetically tractable vertebrate in which environmental or genetic factors can nudge gonads to an ovary or testis fate. Zebrafish gonads develop from bipotential organs in which a few primordial germ cells (PGCs) become oocytes in all juveniles. In some individuals oocytes die and gonads become testes; in others oocytes survive and gonads become ovaries. A key question is the nature of the genetic cascade that causes oocytes to die in some juveniles and survive in others, and how environmental factors alter this cascade. Results suggest the hypothesis that the native SD locus (which we discovered in native strains but is gone from domesticated lab strains) interacts with environmental factors (likely variable nutrition and growth rates) to alter juvenile oocyte survival, which in turn changes the strength of an oocyte-derived signal acting on thegonadal soma to maintain estrogen production, thereby preserving or not preserving oocytes and gonad fate. The aims are: Aim 1 is to learn the molecular genetic basis of the native zebrafish SD locus. Aim 2 is to understand the transcriptomic cascade during SD using: a) a native strain genotyped for genetic sex, b) an environmentally sensitive lab strain that lacks the native SD gene, and c) fish lacking PGCs to identify PGC- derived factors and their downstream targets. Aim 3 is to understand mechanisms of gonadal soma/germ cell reciprocal signaling by learning the roles of the related TGFss signalers Gdf9, Amh, and Gsdf (gonadal soma derived factor), and to discover how the female-specifying signals Wnt4 and Foxl2 act in gonadal development by using induced mutations, many of which we have in hand. Innovation of proposed work includes the identification and unique use of zebrafish possessing and genotyped for the native SD locus; our generally applicable RAD-sex method for identifying sex loci in different populations; the unique mutant and transgenic resources that we make available; and our focus on gonadal soma/germ cell reciprocal signaling in zebrafish sex determination. Outcomes and impact of this work are the potential for a fuller understanding of the molecular genetic nature of the vertebrate sex-balance mechanism, which will increase our appreciation of possible mechanisms underlying increases in human reproductive diseases in developed countries.

描述（由申请人提供）正常的人性性腺发育可能会受到未知的遗传和环境因素的不利影响，导致睾丸不体一代综合征和多囊卵巢综合征。在哺乳动物中，Y染色体基因启动了一个不完全理解的遗传网络，该网络指导性腺成为睾丸，同时竞争基因直接卵巢发育。非哺乳动物脊椎动物使用类似的下游网络，但原发性确定（SD）基因不同。此外，在某些物种中，环境因素（例如温度或营养偏置）。我们的广泛目标是了解遗传因素如何与环境相互作用以使SD平衡点亮。工作重点是斑马鱼，斑马鱼是一种可遗传障碍的脊椎动物，其中环境或遗传因素可以将性腺推向卵巢或睾丸命运。斑马鱼的性腺是由双性器官发展出来的，其中几个原始生殖细胞（PGC）成为所有少年的卵母细胞。在某些人中，卵母细胞死亡，性腺变成睾丸。在其他人中，卵母细胞生存，性腺成为卵巢。一个关键的问题是遗传级联的性质，导致卵母细胞在一些少年中死亡并在其他少年中生存，以及环境因素如何改变这种级联。结果表明，假设是，天然SD基因座（我们在本地菌株中发现，但来自驯化的实验室菌株）与环境因素（可能可变的营养和生长速率）相互作用，以改变少年卵母细胞的存活，从而改变了卵母细胞在雌激素上的卵母细胞信号的强度，而不是维持雌激素，或者维持雌激素的生产。目的是：目标1是学习天然斑马鱼SD基因座的分子遗传基础。 AIM 2是使用：a）用于遗传性别的天然菌株基因分型，b）缺乏天然SD基因的环境敏感的实验室菌株，c）缺乏PGC来识别PGC衍生因子及其下游靶标的FISH。目的3是通过学习相关的TGFSS信号器GDF9，AMH和GSDF（Gonadal Soma衍生的因子）的作用，了解性腺体体/生殖细胞倒数信号传导的机制，并通过使用指定的突变，通过使用指定的MENTINCE，在手段上使用戈纳达尔2在手段上，发现女性指定的信号Wnt4和Foxl2在手段中如何进行。拟议工作的创新包括对本地SD基因座的斑马鱼具有识别和独特的使用；我们通常适用的RAD-SEX方法，用于识别不同人群中的性基因座；我们提供的独特突变和转基因资源；我们的重点是斑马鱼性别确定中的性腺体体/生殖细胞相互信号传导。这项工作的结果和影响是对脊椎动物性增强机制的分子遗传性质的潜在潜力，这将增加我们对发达国家人类生殖疾病潜在机制的欣赏。