RNA LOCALIZATION IN XENOPUS OOCYTES

非洲爪蟾卵母细胞中的 RNA 定位

• 批准号: 2203032
• 负责人: Kimberly L. Mowry
• 金额: $ 16.75万
• 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/2203032
• 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.

在卵子中建立极性可以被描述为最早的步骤 在胚胎模式中。一个单一的细胞，受精卵，含有 最终确定整个生物体的信息。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是 发育潜力是卵子中隔离的分子， 可以具体说明这样的潜力。这是一项由来已久的建议 母性因子在鸡蛋中的定位可以为模式提供依据 在早期胚胎中，虽然有证据表明存在 在许多系统中定位的细胞质决定因素，在任何情况下都不是 了解本地化流程。这项研究提案解决了这个问题。 通过研究特定的母体mRNAs在 青蛙蛋。 这项拟议的研究的目的是了解核糖核酸是如何 被本地化到单元格内的指定区域。非洲爪哇的卵母细胞 为研究这一问题提供了一个很好的模型系统。族长 该系统的实验优势之一是注射速度快 分析定位，并易于获得大量材料 用于生化研究。提出了三条调查路线，以 研究RNA定位：(1)提出了识别和 隔离定位设备的部件。核糖核酸的生物化学 本地化目前还不清楚。通过解决这个问题，新的 应该获得关于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，建议进行以下实验 探讨细胞骨架在RNA定位中的作用。(3)为了 允许了解特定的RNA如何被识别为需要 为了定位，建议进行实验来表征RNA信号 本地化。这些实验可能不仅提供了对如何 发育信号是局部性的，但它在其中可能发挥的作用 细胞类型确定。 目前还不清楚发育信号是如何定位到 在发育早期确定细胞类型。建议进行的实验 这里解决了这个问题，并可能有助于理解如何 在某些疾病中，发育计划会受到干扰。要理解 细胞过程可能出错的手段，首先将是 有必要对正常发展和 差异化。