New conserved candidate for egg activation and early embryogenesis in Drosophila.

果蝇卵激活和早期胚胎发生的新保守候选者。

• 批准号: 8531438
• 负责人: Mariana Federica Wolfner
• 金额: $ 23.21万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8531438
• 关键词: Animal Model; Binding; Biological Models; Calcineurin; Calcium; Calcium Signaling; Cell Cycle; Cells; Chemicals; Development; Developmental Biology; Drosophila genus; Embryo; Embryonic Development; Event; Female; Fertility; Fertilization; Foundations; Future; Genes; Genetic Models; Genetic Transcription; Goals; Infertility; Intracytoplasmic Sperm Injections; Mammals; Mediating; Meiosis; Methods; Mitosis; Mitotic; Modification; Molecular; Molecular Genetics; Oocytes; Oogenesis; Orthologous Gene; Outcome; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Play; Poly(A)+ RNA; Post-Translational Protein Processing; Procedures; Process; Protein Dephosphorylation; Proteins; Proteomics; RNA Interference; Regulation; Regulatory Pathway; Research; Role; Series; Set protein; Source; Staging; Testing; Time; Translating; Vertebrates; Western Blotting; Work; Zinc; base; critical period; design; egg; embryonic protein; fly; novel strategies; public health relevance; research study; screening; text searching; tool

DESCRIPTION (provided by applicant): The transition from egg to embryo is a critical time in development. Since this process is essential for fertility, a better understanding of its mechanisms is important in developmental biology and will also help to optimize ART procedures. It has thus far been very difficult to identify the critical proteins needed for this transition, in part because it is too rapid to involve changes in gene transcription. Instead, numerous lines of evidence argue that this rapid and comprehensive change in cell state is mostly accomplished by post-translational modification of existing proteins. We are focusing on proteins whose phosphorylation state is modulated during egg activation, reasoning that these proteins constitute an enriched pool of molecules important for the egg-to-embryo transition. We propose to use germline-specific RNAi to screen among 258 conserved proteins that our proteomics screens have found to be phospho-modulated during egg activation in Drosophila. By straightforward phenotypic screening that takes advantage of the latest strains and methods for this model organism, we expect to identify the genes that regulate the transition from a mature oocyte to an "activated" egg that can begin development, as well as those genes that initiate its embryogenesis. We will prioritize the order of screening, giving precedence to genes likely to encode upstream regulators of this transition, and to one class of downstream effectors: those likely to regulate the cell cycle as it transitions from meiosis to mitosis. Once we have identified these new players, we will use Western blotting and phosph-proteomics to begin to order their actions. The goal of the proposed research is to discover the critical genes that carry out the important developmental transition from egg to embryo. These genes will permit future studies to dissect in detail the molecular interactions and pathways that carry out egg activation and initiate embryogenesis.

描述（由申请人提供）：从卵子到胚胎的过渡是发育的关键时期。由于这一过程对生育至关重要，因此更好地了解其机制在发育生物学中很重要，也将有助于优化ART程序。到目前为止，很难确定这种转变所需的关键蛋白质，部分原因是它太快了，无法涉及基因转录的变化。相反，许多证据表明，这种快速而全面的细胞状态变化主要是通过对现有蛋白质的翻译后修饰来完成的。我们关注的是在卵子激活过程中磷酸化状态被调节的蛋白质，原因是这些蛋白质构成了一个丰富的分子池，对卵子到胚胎的转变很重要。我们建议使用种系特异性RNAi筛选258个保守蛋白，我们的蛋白质组学筛选发现在果蝇卵激活过程中被磷酸化调节。通过利用最新的菌株和方法对这种模式生物进行直接的表型筛选，我们期望确定调节从成熟卵母细胞到可以开始发育的“激活”卵的转变的基因，以及那些启动其胚胎发生的基因。我们将优先考虑筛选的顺序，优先考虑可能编码这一转变的上游调节因子的基因，以及一类下游效应因子：那些可能调节细胞周期从减数分裂到有丝分裂的基因。一旦我们确定了这些新的参与者，我们将使用Western blotting和磷酸化蛋白质组学开始对它们的行为进行排序。这项研究的目标是发现携带这种疾病的关键基因