Actin in Sea Urchin Eggs: Regulation of the Cytoskeleton

海胆卵中的肌动蛋白：细胞骨架的调节

• 批准号: 8802178
• 负责人: Edward Bonder
• 金额: $ 7万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-01 至 1990-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8802178&HistoricalAwards=false
• 关键词: Actin; Sea; Urchin; Eggs; Regulation

Sea urchin eggs will be used as a system to examine the cellular mechanisms that regulate the actin cytoskeleton. Following fertilization, the egg undergoes definable changes in its actin cytoskeleton in almost perfect synchrony with a series of defined metabolic and ionic changes. The proposed experiments wil examine the in vitro effects of three calcium-sensitive actin binding proteins (egg spectrin, egg 95K, and egg 50K) on actin filament assembly and structure. Polyclonal antibodies monospecific for these proteins will be generated and used for indirect immunofluorescence to examine the spatio-temporal relationship between the actin binding proteins and actin within oocytes, eggs, and embryos. The antibodies will also be used to examine tissue specific expression of the actin binding proteins. If necessary, monoclonal antibodies will be generated to pursue this work. The long term goals of this project are to establish insight into how cells are able to maintain and modulate their actin cytoskeletons with such amazing precision. This is fundamentally important, because the microfilament systems of virtually all eucaryotic cell types are crucial to the proper cytoplasmic and membranous organization (and hence function) of the cells.

海胆卵将被用作检查调节肌动蛋白细胞骨架的细胞机制的系统。受精后，卵子的肌动蛋白细胞骨架发生了可定义的变化，与一系列定义的代谢和离子变化几乎完美同步。 拟议的实验将检查三种钙敏感性肌动蛋白结合蛋白（egg 血影蛋白、egg 95K 和egg 50K）对肌动蛋白丝组装和结构的体外影响。 将生成针对这些蛋白质的单特异性多克隆抗体，并将其用于间接免疫荧光，以检查卵母细胞、卵子和胚胎内肌动蛋白结合蛋白和肌动蛋白之间的时空关系。 这些抗体还将用于检查肌动蛋白结合蛋白的组织特异性表达。 如有必要，将产生单克隆抗体来开展这项工作。 该项目的长期目标是深入了解细胞如何能够以如此惊人的精度维持和调节其肌动蛋白细胞骨架。 这一点非常重要，因为几乎所有真核细胞类型的微丝系统对于细胞的正确细胞质和膜组织（以及功能）都至关重要。