Identify Sex Determination Genes By ENU Mutagenesis

通过 ENU 诱变鉴定性别决定基因

• 批准号: 6575039
• 负责人: James Larry JAMESON
• 金额: $ 50.87万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-27 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6575039
• 关键词: animal breeding; biotechnology; cooperative study; cryopreservation; cytogenetics; ethanol; gene expression; genetic mapping; genetic screening; genotype; gonad disorder; gonads; in situ hybridization; information dissemination; laboratory mouse; mammalian embryology; microarray technology; mutant; online computer; phenotype; polymerase chain reaction; reproductive development; sex chromosomes; sex determination; western blottings

DESCRIPTION (provided by applicant): Sex determination in mammals is governed by a series of genetic switches that influence cell fate and differentiation during critical periods of development. Remarkably, the primordial fetal gonad is bipotential and the precursor gonadal cells can develop along either a male (testis) or female (ovary) pathway, depending on which genes are expressed. The main goal of this project is to identify key genes that regulate gonadal development and phenotypic sex. To this end, the investigators propose to screen mice for sex-reversal, manifest as XY phenotypic females or as XX phenotypic males, taking advantage of the existing Northwestern University Genome Wide ENU Mutagenesis Center. The specific aims of this project are: Aim 1: To screen progeny of ENU-mutagenized mice for sex reversal. Mice will be characterized genetically for the presence or absence of the Y-chromosomal Sry gone. At age 21 days, they will be classified as phenotypically male or female. This screen will include at least 10,000 mice per year. Aim 2: To characterize the gonadal phenotype of mice with sex-reversal. Morphologic, hormonal, histologic, and developmental analyses will be used to characterize the gonadal (testis and ovary) defects associated with sexreversal. Functional analyses will assess spermatogenesis in males or ovulation in females. These initial studies will be followed by more detailed characterizations of altered gene and protein expression using in situ hybridization, quantitative RT-PCR, immunohistology, and western blot studies. Microarray analyses will be used to identify altered genetic pathways, particularly during key stages of gonadal development. Aim 3: To perpetuate germline transmission of mutations associated with sex-reversal and gonadal dysgenesis and to map the genetic locus of the defect. Aim 4: To act as a national resource for mouse mutants by providing access to phenotypic screening analyses "online". In addition to listing identified phenotypes online, putative mutants that are heritable will be cryopreserved and made available to the scientific community directly from the Center and through arrangements with an established national distribution center. Gonadal development provides an excellent opportunity to identify genes involved in differential organogenesis. In addition to providing new information about basic mechanisms that regulate gonad development, these studies are also likely to enhance our understanding of gonadal dysgenesis and infertility in humans.

描述（由申请人提供）：哺乳动物的性别决定由一系列基因开关控制，这些基因开关影响发育关键时期的细胞命运和分化。 值得注意的是，原始胎儿性腺是双能的，前体性腺细胞可以沿着男性（睾丸）或女性（卵巢）途径发育，具体取决于表达的基因。 该项目的主要目标是确定调节性腺发育和表型性别的关键基因。 为此，研究人员建议利用现有的西北大学全基因组 ENU 诱变中心，筛选小鼠的性别逆转，表现为 XY 表型雌性或 XX 表型雄性。 该项目的具体目标是： 目标 1：筛选 ENU 诱变小鼠的后代进行性逆转。 将对小鼠进行基因表征，以确定是否存在 Y 染色体 Sry 缺失。 21 天时，它们的表型将被分为男性或女性。 该屏幕每年至少包含 10,000 只小鼠。 目标 2：表征性逆转小鼠的性腺表型。 形态学、激素、组织学和发育分析将用于表征与性逆转相关的性腺（睾丸和卵巢）缺陷。 功能分析将评估男性的精子发生或女性的排卵。 这些初步研究之后将使用原位杂交、定量 RT-PCR、免疫组织学和蛋白质印迹研究对改变的基因和蛋白质表达进行更详细的表征。 微阵列分析将用于识别改变的遗传途径，特别是在性腺发育的关键阶段。 目标 3：使与性逆转和性腺发育不全相关的突变的种系传播永久存在，并绘制缺陷的遗传位点图谱。 目标 4：通过提供“在线”表型筛选分析，成为小鼠突变体的国家资源。 除了在线列出已识别的表型外，假定的可遗传突变体还将被冷冻保存，并通过中心并通过与已建立的国家分销中心的安排直接提供给科学界。 性腺发育为鉴定参与差异器官发生的基因提供了绝佳的机会。 除了提供有关调节性腺发育的基本机制的新信息外，这些研究还可能增强我们对人类性腺发育不全和不育症的理解。