Establishment and Expression of Polarity in Pelvetia Zygotes

Pelvetia 合子极性的建立和表达

• 批准号: 9207053
• 负责人: Darryl Kropf
• 金额: $ 32.6万
• 依托单位: University of Utah
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-10-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9207053&HistoricalAwards=false
• 关键词: Establishment; Expression; Polarity; Pelvetia; Zygotes

This work will investigate the mechanism(s) by which polarity is established and expressed in fertilized eggs of fucoid algae. Much of the work will focus on the role of the cytoskeleton. Three major objectives concerning the cytoskeleton will be pursued. (1) An in-depth characterization of the structure of microfilament and microtubule networks, and especially interactions between these networks. These experiments will rely on confocal microscopy. Preliminary investigations of actin-binding proteins and microtubule-associated proteins will be conducted using biochemical approaches and specific antibody probes. (2) Dynamic rearrangements of cortical microfilaments and endoplasmic microtubules will be investigated in real time by microinjecting fluorescent probes into living cells. Inhibitors will be use to investigate the function of cytoskeletal rearrangements in embryogenesis. (3) Mechanisms of intracellular transport will also be examined using microinjection. Two types of intracellular transport will be studied, pronuclear migration and secretory vesicle transport. The second part of the research will develop the use of protoplasts as an experimental tools in two lines of research. First, protoplasts of the rhizoid cell provide a unique opportunity to investigate the loss of polarity, and will be investigated as models of 'depolarization'. Second, molecular approaches to studying polarity will be developed beginning with production and initial characterization of a cDNA library from protoplasts.

这项工作将研究岩藻类藻类受精卵极性建立和表达的机制。 大部分工作将集中在细胞骨架的作用上。 将追求有关细胞骨架的三个主要目标。 （1）深入表征微丝和微管网络的结构，特别是这些网络之间的相互作用。 这些实验将依赖于共焦显微镜。 将使用生化方法和特异性抗体探针对肌动蛋白结合蛋白和微管相关蛋白进行初步研究。 （2）通过将荧光探针显微注射到活细胞中，实时研究皮质微丝和内质微管的动态重排。 抑制剂将用于研究胚胎发生中细胞骨架重排的功能。 (3) 还将使用显微注射来检查细胞内运输的机制。 将研究两种类型的细胞内运输：原核迁移和分泌囊泡运输。 该研究的第二部分将开发使用原生质体作为两个研究领域的实验工具。 首先，假根细胞的原生质体提供了研究极性丧失的独特机会，并将作为“去极化”模型进行研究。 其次，将从原生质体 cDNA 文库的产生和初步表征开始，开发研究极性的分子方法。