FIBROBLAST

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们已经基于边缘的形态学和运动学以及存在的蛋白质定义了在细胞边缘发生的许多不同类型的运动性（Dubin-Thaler，提交;（Dobereiner et al.，2005））。 由于前缘通常非常薄（<200 nm），因此应该可以在冷冻电镜中观察前缘处复合物的分子水平形态。 许多复合物已在体外可视化，因此，它可能是可能的，以确定在体内的复合物，这将使一个更好地了解不同类型的运动在分子水平上。 特别是，我们建议准备胞质体（细胞质的去核部分）和纤连蛋白表面上的传播特性。 然后，当我们在光学显微镜下记录细胞质时，细胞质就会扩散。 这将使我们能够确定正在发生的运动类型。 在适当的时候，样品将用甲醛固定，随后将在非常薄的玻璃状冰层中冷冻。 或者，允许样品扩散一段规定的时间，然后冷冻而不固定。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have defined a number of different types of motility that occur at the edges of cells based upon the morphology and kinematics of the edge as well as the proteins present (Dubin-Thaler, submitted; (Dobereiner et al., 2005)). Because the leading edge is typically very thin (<200 nm), it should be possible to visualize the molecular-level morphology of the complexes at the leading edge in the cryo-em. Many of the complexes have been visualized in vitro; therefore, it may be possible to identify the in vivo complexes, which will enable a much better understanding of the different types of motility at the molecular level. In particular, we propose to prepare cytoplasts (enucleated portions of cytoplasm) and characterize their spreading on fibronectin surfaces. Cytoplasts would then be spread as we recorded them in the light microscope. This would enable us to define the type of motility that was occurring. At the appropriate time, the sample would be fixed with formaldehyde and would subsequently be frozen in a very thin layer of vitreous ice. Alternatively, the samples would be allowed to spread for a defined period and then frozen without fixation.