Role of the primary cillium in glomerular podocyte cells

描述(由申请人提供)：本研究方案的长期目标是确定足细胞中初级纤毛的作用。在蛋白尿肾病的发展过程中，足细胞的外观会发生改变，例如假性囊肿和足突消失。肾集合管细胞已被证明在体外通过顶端质膜的初级纤毛感知液体流动。由于足细胞在解剖学上位于高液体流动的区域，一种假设是足细胞的初级纤毛感知液体流动。这一主要假设被整合为四个具体目标。第一个目标将使用标准成像技术来确定足细胞纤毛的长度、频率和微管排列。第二个目标将解决足细胞可以通过监测细胞内钙离子的变化来感知液体流动的假设。细胞内钙离子将使用标准的灌流设备和显微镜进行监测。第三个目标将使用初级纤毛的生物物理和生化操作来确定初级纤毛是否是一个流量感受器。第四个目标是确定哪些细胞内和细胞外的钙通道，如多囊蛋白和瞬时受体电位通道(TRPC)，参与了流动诱导的反应。CA++。

DESCRIPTION (provided by applicant): The long term objective of this research proposal is to determine the role of the primary cilium in podocytes. In the progression of proteinuric renal diseases, there are alterations in podocyte appearance, e.g. pseudocysts and foot process effacement. Renal collecting duct cells have now been shown to sense fluid flow by the apical plasma membrane's primary cilium in vitro. Since podocytes are anatomically located in an area of high fluid flow, one hypothesis would be that the podocyte's primary cilium senses fluid flow. This major hypothesis is integrated into four specific aims. The first aim will employ standard imaging techniques to determine the podocyte cilia in terms of length, frequency, and microtubular arrangement. The second aim will address the hypothesis that podocytes can sense fluid flow by monitoring changes in intracellular Ca++. Intracellular calcium will be monitored using a standard perfusion apparatus and microscope. The third aim will use biophysical and biochemical manipulation of the primary cilium to determine if the primary cilium is a flow sensing organelle. The fourth aim will identify which intracellular and extracellular calcium channels are involved, e.g. polycystins and transient receptor potential channels (TRPC), in the flow induced response. Ca++.