Membrane Dynamics Underlying Kv2.1 Cluster Formation

在这个项目中，PI将研究Kv2.1通道簇的调节，这些通道簇可能在大脑中发挥几种重要的生物学作用。目前，Kv2.1簇的调节和维持机制尚不清楚。该项目侧重于Kv2.1电压门控K+通道细胞表面动力学的生物物理学，特别强调活细胞中的细胞膜相互作用。这项研究的总体目标是提高我们对皮质细胞骨架功能性地形成扩散限制栅栏的机制的理解，该扩散限制栅栏选择性地限制Kv2.1通道的亚群。具体的研究目标是：（1）表征培养神经元的各种表面区域内的成簇和非成簇通道的动力学，（2）测量皮质细胞骨架和筏微区对Kv2.1通道动力学的影响，（3）构建显微镜以实现高速粒子跟踪和光镊，（4）确定Kv2.1簇在细胞表面形成的机制。这项实验工作将导致膜蛋白动力学的分子机制的独特见解。该项目的重点是研究Kv2.1的聚类调节，动力学和与细胞骨架的相互作用，这可能会导致通过增强神经保护方法改善急性缺血性卒中的治疗。该研究计划将通过建立用于教育目的的光镊装置与教育部分相结合。代表性不足的学生将在少数民族会议上和通过科罗拉多州立大学的少数民族妇女工程方案招募。该项目由国家科学基金会物理和分子细胞生物学部门联合赞助。

In this project the PIs will study the regulation of Kv2.1 channel clusters that may play several important biological roles in the brain. Currently, the mechanism by which clusters Kv2.1 are regulated and maintained is unknown. The project focuses on the biophysics of Kv2.1 voltage-gated K+ channel cell surface dynamics with particular emphasis on cytoskeleton-membrane interactions in live cells. The overall goal of the research is to improve our understanding of the mechanism by which the cortical cytoskeleton functionally forms a diffusion limiting fence that selectively corrals a sub-population of Kv2.1 channels. The specific research aims are: (1) characterize the dynamics of clustered and non-clustered channels within various surface regions of cultured neurons, (2) measure the influence of the cortical cytoskeleton and raft microdomains on Kv2.1 channel dynamics, (3) build a microscope to implement high-speed particle tracking and optical tweezers, and (4) determine the mechanism that forms Kv2.1 clusters on the cell surface. This experimental work will lead to unique insights in the molecular mechanism of membrane protein dynamics. The project focuses on investigating Kv2.1 clustering regulation, dynamics and interactions with the cytoskeleton which may lead to improved treatments for acute ischemic stroke through enhanced neuro-protective approaches. The research program will be integrated with an educational component by building an optical tweezers setup for educational purposes. Under-represented students will be recruited at minority meetings and through a Women & Minorities in Engineering Program at Colorado State University.The project is jointly sponsored by the Physics and the Molecular Cell Biology Divisions at NSF.