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.

卵中的极性建立可以描述为最早的步骤 在胚胎图案中。 一个细胞，受精卵，包含 最终将指定整个生物体的信息。 作为鸡蛋 在开发的早期阶段分裂，不同的细胞获取 不同的命运。 考虑到这种差异的一种机制 发育潜力是卵中分子的隔离， 可以指定这种潜力。 这是一项长期的提议 鸡蛋中母体因素的定位可以为模式提供基础 在早期的胚胎中，虽然有证据表明存在 在许多系统中 理解本地化过程。 这项研究建议解决了这一点 通过研究特定母体mRNA在 青蛙鸡蛋。 拟议研究的目的是了解RNA的手段 位于单元内的指定区域。 爪蟾卵母细胞 提供了一个出色的模型系统来研究此问题。 首席 在该系统中的实验优势中有快速注射 测定本地化，并轻松获得大量材料 用于生化研究。 提出了三条调查线 研究RNA定位：（1）提出了实验以识别和 隔离设备的分离成分。 RNA的生物化学 目前尚不了解本地化。 通过解决这个问题，新 应获得有关涉及的分子设备的信息 在将大分子转移到卵母细胞细胞质的定义区域时。 （2） 为了解决某些机械性问题，提出了实验 探测细胞骨架在RNA定位中的作用。 （3）为了 允许了解特定RNA的认可 本地化，提出了实验以表征RNA信号 本土化。 这些实验不仅可以为如何 发育信号是本地化的，但也可能在 细胞类型的确定。 目前，尚不了解发展信号如何本地化 在早期开发过程中确定细胞类型。 提出了实验 在这里解决这个问题，并可能有助于了解 发展计划在某些疾病中受到干扰。 理解 蜂窝过程可能会出现问题的方式，首先是 对正常发展的基本理解和 分化。