Actin in Sea Urchin Eggs: Regulation of the Cytoskeleton

海胆卵将被用作一个系统， 调节肌动蛋白细胞骨架的机制。以下 受精后，卵子在其肌动蛋白中发生了可确定变化， 细胞骨架几乎与一系列定义的 代谢和离子变化。 拟议的实验将检查 三种钙敏感肌动蛋白结合蛋白（卵 血影蛋白、卵95K和卵50K）对肌动蛋白丝组装的影响， 结构 对这些蛋白质具有单特异性的多克隆抗体 产生并用于间接免疫荧光，以检查 肌动蛋白结合蛋白之间的时空关系 卵母细胞、卵子和胚胎内的肌动蛋白。 抗体也将 用于检测肌动蛋白结合的组织特异性表达 proteins. 如有必要，将产生单克隆抗体， 追求这项工作。 这个项目的长期目标是建立洞察如何 细胞能够维持和调节它们的肌动蛋白细胞骨架， 如此惊人的精确度 这一点至关重要，因为 几乎所有真核细胞类型的微丝系统都是 对适当的细胞质和膜组织至关重要（和 功能）。

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.