FIBROBLAST

成纤维细胞

• 批准号: 7953799
• 负责人: MICHAEL Patrick SHEETZ
• 金额: $ 0.87万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7953799
• 关键词: Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Electron Microscopy; Fibroblasts; Fibronectins; Formaldehyde; Freezing; Funding; Grant; Ice; In Vitro; Institution; Light Microscope; Molecular; Morphology; Proteins; Research; Research Personnel; Resources; Sampling; Source; Surface; Time; United States National Institutes of Health; base; cell motility; in vivo; kinematics; macromolecule; sample fixation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我们已经定义了基于边缘的形态和运动学以及存在的蛋白质（Dubin-thaler，提交的；（Dobereiner等，2005）），在细胞边缘发生了多种不同类型的运动。 由于前缘通常非常薄（<200 nm），因此应该有可能可视化冷冻EM中前缘的复合物的分子水平形态。 许多复合物已在体外可视化；因此，可能可以识别体内复合物，这将使人们能够更好地了解分子水平的不同类型的运动。 特别是，我们建议制备细胞体（细胞质的柠檬核部分），并表征它们在纤连蛋白表面上的扩散。 然后，当我们将其记录在光学显微镜中时，细胞体将扩散。 这将使我们能够定义发生的运动类型。 在适当的时间，样品将用甲醛固定，随后将在非常薄的玻璃冰中冷冻。 或者，将允许样品在定义的时期扩散，然后冷冻而无需固定。