Supplement for ZetaSizer instrument

联系PD/PI：卡尔弗特，彼得·迪恩 项目摘要/摘要 这项建议的目的是确定光感受器蛋白的机制。 分隔化。视网膜光感受器是一种极化神经元，其主要功能是 包括接收和发送信号，被划分为离散亚蜂窝 域名。分区对于正常的光感受器活动和视力下降或 失明是由于蛋白质分离不当造成的。尽管它们很重要，但这些机制 光感受器或任何其他神经元中潜在的蛋白质区隔作用仍然很差。 明白了。理解区划的关键是生物物理特性 光感受器细胞质，蛋白质的生物物理性质 以及驱动蛋白质积累的力量，而不是显著的 浓度梯度，进入特定的隔间。我们发现了一个基本的 生物物理机制可能是蛋白质运输和电子分离的中心 活性细胞：光感受器产生的电场内带电蛋白质的运输 神经元活动。我们将这一新机制称为轴向动态电泳蛋白 运输(熟练)。我们将使用最先进的活细胞荧光成像工具 我们的实验室，强大的非洲爪哇转基因和基因编辑技术，以及尖端的 生物化学和细胞生物学方法，以实现以下目标： 目的1：定位杆状感光细胞轴向胞质电场EAX。 目的2：确定ADEPT对光感受器蛋白转运的影响 活的光感受器的区隔。 目的3：确定arrestin相互作用的位置及其对其分布的影响 以及活的杆状和圆锥体的动力学。 项目摘要/摘要第7页

Contact PD/PI: Calvert, Peter Deane PROJECT SUMMARY/ABSTRACT The objectives of this proposal are to determine the mechanisms of photoreceptor protein compartmentalization. Retinal photoreceptors are polarized neurons whose major functions, include receiving and transmitting signals, are compartmentalized into discrete subcellular domains. Compartmentalization is critical for normal photoreceptor activity and reduced vision or blindness result from improper segregation of proteins. Despite their importance, the mechanisms underlying protein compartmentalization in photoreceptors, or any other neuron, remain poorly understood. Essential for understanding compartmentalization are the biophysical properties of the photoreceptor cytoplasm, the biophysical properties of the proteins that are destined to be compartmentalized and the forces that drive accumulation of proteins, against significant concentration gradients, into the specific compartments. We have uncovered a fundamental biophysical mechanism that may be central to protein transport and segregation in all electrically active cells: transport of charged proteins within the electrical field generated by the photoreceptor neuronal activity. We call this novel mechanism axial dynamic electrophoretic protein transport (ADEPT). We will use state of the art live cell fluorescence imaging tools developed in our lab, powerful transgenic and gene editing techniques in Xenopus, and sophisticated biochemical and cell biological approaches to address the following aims: Aim 1: Map the axial cytoplasmic electric field, Eax, in rod photoreceptors. Aim 2: Determine the impact of ADEPT on photoreceptor protein transport and compartmentalization in living photoreceptors. Aim 3: Determine the locations and influence of Arrestin interactions on their distributions and dynamics in living rods and cones. Project Summary/Abstract Page 7