Actin in Sea Urchin Eggs: Regulation of the Cytoskeleton

海胆卵将被用作一个系统来研究调节肌动蛋白细胞骨架的细胞机制。受精后，卵子在其肌动蛋白细胞骨架上经历了可定义的变化，几乎与一系列确定的代谢和离子变化完全同步。本实验将检测三种钙敏感的肌动蛋白结合蛋白(鸡蛋血影蛋白、鸡蛋95K和鸡蛋50K)在体外对肌动蛋白细丝组装和结构的影响。将产生针对这些蛋白的单特异性多克隆抗体，并用于间接免疫荧光，以检测卵子、鸡蛋和胚胎中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测肌动蛋白结合蛋白的组织特异性表达。如果有必要，将产生克隆抗体来进行这项工作。该项目的长期目标是深入了解细胞如何能够如此精确地维持和调节其肌动蛋白细胞骨架。这是非常重要的，因为几乎所有真核细胞类型的微丝系统对细胞的适当的细胞质和膜组织(从而功能)是至关重要的。

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.