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扮演几个重要的生物学角色。目前，集群Kv2.1的调节和维持机制尚不清楚。该项目专注于Kv2.1电压门控K+通道细胞表面动力学的生物物理学，特别强调活细胞中细胞骨架-膜的相互作用。这项研究的总体目标是提高我们对皮质细胞骨架在功能上形成扩散限制屏障的机制的理解，该屏障选择性地围栏Kv2.1通道的亚群。具体的研究目标是：(1)表征培养神经元不同表面区内聚集和非聚集通道的动力学；(2)测量皮质细胞骨架和RAFT微域对Kv2.1通道动力学的影响；(3)建立显微镜以实现高速粒子跟踪和光钳；(4)确定细胞表面形成Kv2.1簇的机制。这项实验工作将对膜蛋白动力学的分子机制产生独特的见解。该项目的重点是研究Kv2.1的聚集调节、动力学以及与细胞骨架的相互作用，这可能会通过增强神经保护方法来改进急性缺血性中风的治疗。该研究计划将与一个教育组成部分相结合，建立一个用于教育目的的光学镊子装置。代表不足的学生将在少数族裔会议上招募，并通过科罗拉多州立大学的女性和少数族裔工程项目来招募。该项目由NSF的物理学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.