US-United Kingdom Cooperative Research: Assembly of the Neuronal Cytoskeleton

该奖项将支持科罗拉多州立大学的James Bamburg博士和伦敦医学研究委员会细胞生物物理部的Dennis Bray博士之间的合作研究。在这个项目中，Bamburg博士和Bray博士将使用荧光和电子显微镜研究肌动蛋白组装到神经元皮质的位置。肌动蛋白细丝是神经元生长锥的主要细胞骨架成分，构成微棘的核心结构，即从生长锥延伸出来的伪足，产生定向生长所需的张力。Bamburg博士和Bray博士计划确定肌动蛋白和其他传输到轴突皮质的蛋白质之间可能的复合体。拟议的实验应该确定肌动蛋白亚单位是否在运输过程中在轴突中的细丝群体之间交换，以及轴突的皮质肌动蛋白层是否来自生长锥体或轴突的沉积。在项目过程中，班伯格博士将准备用于显微注射的蛋白质；布雷博士将准备针对荧光标记的抗体。微量注射研究将在两个实验室进行。拟议的研究非常适合这两个实验室之间的合作。Bray博士在神经元培养方法的开发、神经元细胞蛋白抗体的生产以及免疫细胞化学方法在神经元内其他蛋白质定位方面的应用方面一直是领导者。Bamburg博士在体外分离和鉴定与细胞骨架结构有关的蛋白质以及调节细胞骨架组件的组装方面一直很活跃。

This award will support collaborative research between Dr. James Bamburg, Colorado State University, and Dr. Dennis Bray, Cell Biophysics Unit, Medical Research Coucil, London. In this project Dr. Bamburg and Dr. Bray will investigate the sites of assembly of actin into the neuronal cortex using fluorescence and electron microscopy. Actin filaments are the major cytoskeletal element of the neuronal growth cone and comprise the core structure of microspikes, pseudopodia which extend from the growth cone and which generate the tension required for directed neurite growth. Drs. Bamburg and Bray plan to identify putative complexes between actin and other proteins in transit to the axonal cortex. The proposed experiments should determine if actin subunits exchange between the filament population in the axon during transport, and if the cortical actin layer of the axon arises from deposition at the growth cone or the shaft. During the course of the project, Dr. Bamburg will prepare proteins for microinjection; Dr. Bray will prepare antibodies against the fluorescent labels. The microinjections studies will be carried out in both laboratories. The proposed studies are well suited for collaboration between these two laboratories. Dr. Bray has been a leader in the development of neuronal culture methods, production of antibodies to neuronal cell proteins, and the application of immunocytochemical methods for other localization of proteins within neurons. Dr. Bamburg has been active in isolation and characterization in vitro of proteins ________ involved in cytoskeletal structures and in regulating assembly of cytoskeletal components.