RNA LOCALIZATION IN XENOPUS OOCYTES

非洲爪蟾卵母细胞中的 RNA 定位

• 批准号: 2855615
• 负责人: Kimberly L. Mowry
• 金额: $ 10万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2855615
• 关键词: DNA binding protein; DNA footprinting; RNA binding protein; SDS polyacrylamide gel electrophoresis; Xenopus oocyte; affinity chromatography; biological signal transduction; cell type; cellular polarity; crosslink; cytoskeleton; early embryonic stage; gel mobility shift assay; genetic regulatory element; histogenesis; laboratory rabbit; messenger RNA; microinjections; polymerase chain reaction; protein sequence; radiotracer; site directed mutagenesis; ultraviolet radiation

Establishment of polarity in the egg can be described as the earliest step in embryonic patterning. A single cell, the fertilized egg, contains the information that will ultimately specify the entire organism. As the egg divides during the early stages of development, different cells acquire different fates. One mechanism to account for this difference in developmental potential is the sequestering of molecules in the egg that could specify such potential. It is a long-standing proposal that localization of maternal factors in eggs can provide the basis for pattern in the early embryo, and while there is evidence for the existence of localized cytoplasmic determinants in many systems, in no case is the localization process understood. This research proposal addresses this problem by investigating the localization of specific maternal mRNAs in the frog egg. The aim of the proposed research is to understand the means by which RNA is localized to specified regions within a cell. The Xenopus oocyte provides an excellent model system in which to study this problem. Chief among the experimental advantages in this system are a rapid injection assay for localization, and ease in obtaining large amounts of material for biochemical studies. Three lines of investigation are proposed to study RNA localization: (1) Experiments are proposed to identify and isolate components of the localization apparatus. The biochemistry of RNA localization is not currently understood. By addressing this issue, new information should be gained concerning the molecular apparatus involved in moving macromolecules to defined regions of the oocyte cytoplasm. (2) To address certain mechanistic questions, experiments are proposed to probe the role of the cytoskeleton in RNA localization. (3) In order to allow understanding of how particular RNAs are recognized as requiring localization, experiments are proposed to characterize the RNA signal for localization. These experiments may provide insights not only into how developmental signals are localized, but also the role this may play in cell type determination. Currently, it is not understood how developmental signals are localized to determine cell type during early development. The experiments proposed here address that problem, and may contribute towards understanding how developmental programs are disrupted in certain diseases. To understand the means by which cellular processes can go awry, it will first be necessary to gain a basic understanding of normal development and differentiation.

鸡蛋极性的建立可以说是最早的一步 在胚胎模式中。 单个细胞，即受精卵，含有 最终将指定整个有机体的信息。 就像鸡蛋一样 在发育的早期阶段分裂，不同的细胞获得 不同的命运。 解释这种差异的一种机制 发育潜力是卵子中分子的隔离 可以具体说明这种潜力。 这是一项由来已久的提议 母体因素在卵中的定位可以为模式提供基础 在早期胚胎中，虽然有证据表明存在 许多系统中的局部细胞质决定因素，在任何情况下都不是 了解本地化过程。 本研究提案解决了这个问题 通过研究特定母体 mRNA 的定位来解决问题 青蛙蛋。 拟议研究的目的是了解 RNA 的作用方式 定位于细胞内的指定区域。 爪蟾卵母细胞 提供了一个优秀的模型系统来研究这个问题。 首席 该系统的实验优势之一是快速注射 定位分析，并轻松获得大量材料 用于生化研究。 建议开展三项调查 研究 RNA 定位：（1）建议进行实验来识别和 隔离定位装置的组件。 RNA的生物化学 目前尚不了解本地化。 通过解决这个问题，新 应获得有关所涉及的分子装置的信息 将大分子移动到卵母细胞细胞质的特定区域。 (2) 为了解决某些机械问题，建议进行实验 探究细胞骨架在 RNA 定位中的作用。 (3) 为了 允许了解特定 RNA 是如何被识别为需要的 定位，建议进行实验来表征 RNA 信号 本土化。 这些实验不仅可以提供关于如何 发育信号是局部的，但它可能发挥的作用 细胞类型测定。 目前，尚不清楚发育信号如何定位到 确定早期发育期间的细胞类型。 提出的实验 这里解决这个问题，并可能有助于理解如何 某些疾病会扰乱发育程序。 要了解 细胞过程可能出错的方式，首先是 获得正常发育的基本了解所必需的 差异化。