APICAL IF MEDIATE POST-ISCHEMIA RECOVERY IN EPITHELIA

描述(摘自申请者的摘要)：缺血是一个主要原因 急性肾功能衰竭(ARF)，也是导致功能障碍的重要原因 其他上皮性器官，如肝脏和肠道，尤其是在 移植。上皮细胞的缺血性损伤具有复杂的多发性 效果。然而，人们对质膜极性丧失达成了共识 导致细胞骨架解体的主要原因之一是 上皮功能衰竭的病理生理机制。最近一次 这一领域的研究主要集中在对肌动蛋白细胞骨架的影响上， 尽管已有报道称，微管在 再灌流。申请人的实验室最近表明，根尖网络 中间丝在组织中起着重要的作用 顶极，通过维持F-肌动蛋白微绒毛核心和 微管的组织。在这方面，他刚刚报告说， 心尖IF网络维持微管组织中心的地位 (MTOC)约束它。初步数据表明，中频网络是 在ATP耗尽期间保持稳定，且在ATP耗尽期间MTOC从其上分离。 此外，利用CK8基因的转基因敲除，他已经证明 末端网确实是顶极所必需的。这些数据 为假设顶端IF网络是什么提供了基础 负责ATP耗尽后MTOC的重新本地化 IF相关蛋白的功能，或者直接通过结合发挥作用 细胞角蛋白。为了验证这一假设，具体目标是： 1)研究IF蛋白对细胞骨架再极化的特异性。 三磷酸腺苷消耗。 2)鉴定与微管极化恢复有关的蛋白质 三磷酸腺苷诱导的内皮细胞缺血性损伤及其分子变化 耗尽。 这个项目的长期目标是确定分子机制。 调节MTOC从中间纤维中分离，以确定可能的 替代治疗方法来阻断它们，从而保护上皮细胞 防止缺血损伤或促进其恢复。

DESCRIPTION (Adapted from the Applicant's Abstract): Ischemia is a major cause of acute renal failure (ARF), and also an important cause of dysfunction in other epithelial organs such as liver and intestine, especially after transplantation. The ischemic injury in epithelial cells has complex multiple effects. However, there is a consensus in that loss of plasma membrane polarity resulting from the disorganization of the cytoskeleton is one of the major pathophysiological mechanisms for epithelial function failure. Most recent research in this area has focused in the effects on the actin cytoskeleton, although it has been reported that microtubules are also disorganized during reperfusion. The applicant's lab has recently shown that the apical network of intermediate filaments plays a substantial role in the organization of the apical pole, by maintaining the stability of F-actin microvillus cores and the organization of microtubules. In this regard, he has just reported that the apical IF network maintains the position of Microtubule Organizing Centers (MTOC) that bind it. The preliminary data indicate that the IF network is stable during ATP depletion and that MTOC detach from it during ATP depletion. Furthermore, using a transgenic knock-out of the CK8 gene, he have demonstrated that the terminal web is indeed required for apical polarity. These data provide the foundation for a hypothesis where the apical IF network is responsible for the re-localization of MTOC after ATP depletion through the function of IF-associated proteins, or, alternatively, directly through binding to the cytokeratins. To test this hypothesis, the specific aims are: 1) Study the specificity of IF proteins to re-polarize the cytoskeleton after ATP depletion. 2) Characterize the proteins involved in regaining microtubule polarity after an ischemic injury in epithelia cells and their molecular changes during ATP depletion. The long term goals of this project are to identify the molecular mechanisms mediating the detachment of MTOC from intermediate filaments to define possible therapeutic alternatives to block them and, therefore, protect epithelial cells from ischemic injury or promote their recovery.