MICROTUBULES AND REGULATION OF FOCAL CONTACTS

焦点接触是细胞的主要结构特化 膜和皮层肌动蛋白细胞骨架负责粘附 培养细胞的基质;然而，很少有人知道， 控制其发育的细胞骨架机制。初步数据， 由莫斯科的小组用微管解聚药物获得 这导致了微管系统可能控制 通过对国家的管制， 肌动蛋白皮质。 反过来，皮层产生的收缩力 其作用于病灶细胞膜上的细胞粘附蛋白， 接触受到影响。 本项目的目标是研究 微管系统的发展重点接触。 具体目标 包括对震源区结构和动力学变化的分析， 干预后培养的成纤维细胞中的接触特异性改变 微管系统的结构和功能。 其中包括暴露 细胞对药物解聚或稳定微管的反应 注射微管运动蛋白驱动蛋白抗体。两 补充技术将用于评估焦点状态 接触：干涉反射显微镜，以揭示这些接触， 活细胞和免疫细胞化学方法来鉴定蛋白质 特异性定位于病灶接触和其他细胞骨架 proteins. 为了测试皮层收缩力的变化， 诱导的微管特异性试剂，并将这些变化 与那些焦点接触力测量方法的发展 将使用查佩尔山实验室中的。 计划中的实验将有助于更好地了解 细胞骨架控制细胞基质形成的机制 粘连 该项目将探讨改建的性质， 细胞膜-基质的关系，负责病理 肿瘤细胞的粘附性和运动性。

Focal contacts are the main structural specializations of the cell membrane and of cortical actin cytoskeleton responsible for the adhesion of cultured cells to the substratum; however, little is known about the cytoskeletal mechanisms controlling their development. Preliminary data, obtained by the group in Moscow, with microtubule-depolymerizing drugs resulted in the hypothesis that the microtubule system may control the organization of focal contacts via the regulation of the state of the actin cortex. In turn, the contractile forces developed by the cortex which act on the cytoskeleton-attached membrane proteins of the focal contacts are affected. The goal of this project is to study the role of microtubular system in the development of focal contacts. Specific aims include the analysis of the alterations of structure and dynamics of focal contacts in cultured fibroblasts after interventions specifically changing the structure and function of microtubular system. These include exposure of cells to the drugs depolymerizing or stabilizing microtubules as well as injection of antibody to microtubule motor protein, kinesin. Two complementary technologies will be used to assess the state of focal contacts: interference-reflection microscopy to reveal these contacts in living cells and immunocytochemical methods to identify proteins specifically localized in the focal contacts and for other cytoskeletal proteins. To test the alterations of contractile forces in the cortex induced by microtubule-specific agents and to correlate these alterations with those of focal contacts the methods of force measurements developed in the Chapel Hill laboratory will be used. The planned experiments will lead to better understanding of the mechanisms of cytoskeletal control of the formation of cell-matrix adhesions. The project will explore the nature of alterations in the cytoskeleton-membrane-matrix relationships responsible for pathological adhesion and motility of neoplastic cells.