Construction and stability of photoreceptor outer segment discs

项目总结 这项建议的目的是确定光感受器盘膜的机制 详述。视盘膜是视觉的中心细胞器，吸收光子 随后就有了光转导。毁灭性的致盲疾病表现为有序的中断 外节段盘的堆叠，表明维持这种顺序对光感受器来说是必不可少的 健康和功能。椎间盘形成的一个关键因素是盘内的紧密并列 膜面，它们之间只有几个纳米的距离。这个同义词是非常有力的 并且因此产生必须克服的能量障碍以形成盘。查找 弄清这种能量障碍是如何克服的，对于理解杆和锥体外部是很重要的。 分段精细化和稳定性。我们将使用最先进的活细胞荧光成像工具 我们实验室开发的强大的非洲爪哇转基因和基因编辑技术 生物化学和细胞生物学方法，以实现以下目标： 目的1：确定视蛋白二聚体或更高阶低聚物在形成和维持中的作用 杆状和锥形感光器盘。 目的2：确定视蛋白N-糖基化在视杆和视锥的生成和维持中的作用 光感受器光盘。 目的3：确定膜电荷屏蔽对杆状和锥状结构的贡献 圆盘的形成和稳定性。

Project summary The objectives of this proposal are to determine the mechanisms of photoreceptor disc membrane elaboration. Disc membranes are the central organelles for vision where photons are absorbed and phototransduction ensues. Devastating blinding disease manifest as disruptions in orderly stacks of outer segment discs, showing that maintaining this order is essential to photoreceptor health and function. A key element to disc formation is the close juxtaposition of the intradiscal membrane faces, which are only a few nanometers apart. This apposition is highly energetically unfavorable and thus produces an energy barrier that must be overcome for discs to form. Finding out how this energy barrier is overcome is important for understanding rod and cone outer segment elaboration and stability. 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 biochemical and cell biological approaches to address the following aims: Aim 1: Determine the roles of opsin dimers or higher order oligomers in forming and maintaining rod and cone photoreceptor discs. Aim 2: Determine the roles of opsin N-glycosylation in generating and maintaining rod and cone photoreceptor discs. Aim 3: Determine the contributions of membrane charge shielding by electrolytes to rod and cone disc formation and stability.