CEK7 RAGS RECEPTOR LIGAND RELATIONSHIP IN RETINA AND TEC

这项提案调查了一个推定的排斥因素，破布，和 内源性鸡顶盖蛋白被认为参与了 视网膜神经节细胞发育过程中的指导。在轴突期间 在体外生长，Drescher已经表明轴突可以避免生长到 抹布上标有细胞。候选人提议测试是否 RAGS的内源性受体是CEK7，一种EPH受体酪氨酸 鸡视网膜中有一种激活剂。实验进行得很好- 构思时有明确的概念基础。这些技术很容易掌握。 可供赞助商使用，并且所有必要的试剂都已 已创建。根据与其他物种的同源性，似乎很可能 这项建议将创建非常有用的发展系统，在其中 研究轴突引导。演示RAGS/CEK7功能 相互作用将增加我们对抑制因素的扩展知识 阻止视神经移植后的功能性再生 神经损伤。 该项目分为两个具体目标。首先，候选人 提出用来演示配体和 受体，并确定哪些受体亚型可以与 破布。将使用RAG和CEK7的重组来源。激活 受体的活性将通过自身磷酸化来测量。第二 AIM将调查RAGS/CEK7相互作用是否真的 使用真正的小鸡视网膜神经节细胞令人厌恶。以显示 排斥相互作用的特殊性，CEK7将被灭活 生色团辅助激光灭活。CEK7的流动率 将确定激光灭活后的视网膜测试 轴突引导可以研究轴突在存在或不存在时的生长 激活的CEK7。条纹分析将显示不活跃的CEK7轴突 朝向碎布细胞生长，活跃的CEK7轴突避开碎布轴突。

This proposal investigates a putative repulsive factor, RAGS, an endogenous chick tectal protein that is believed to be involved in the guidance of retinal ganglion cells during development. During axon growth in vitro, Drescher has shown the axons avoid growing into RAGS labeled cells. The candidate proposes to tested whether the endogenous receptor for RAGS is CEK7, an EPH receptor tyrosine kinase shown to be in chick retina. The experiments are well- conceived with a clear conceptual basis. The techniques are readily available to the sponsor and all necessary reagents have been created. Based on homologies with other species, it seems likely that this proposal will created very useful developmental system in which to study axon guidance. Demonstrating a RAGS/CEK7 functional interaction will add to our expanding knowledge of inhibitory factors that prevent functional regeneration of transplantation following optic nerve damage. The project is divided into two specific aims. First, the candidate proposed to demonstrate the interaction between the ligand and receptor, and to determine which of the receptor isoforms can bind to RAGS. Recombinant sources of RAGS and CEK7 will be used. Activation of the receptor will be measured by autophosphorylation. The second aim will investigate whether the RAGS/CEK7 interaction is actually repulsive using actual chick retinal ganglion cells. To show the specificity of a repulsive interaction, CEK7 will be inactivated by chromophore-assisted laser inactivation. The CEK7 turnover rate following laser inactivation will be determined so that tests of retinal axon guidance can study axon growth in the presence or absence of active CEK7. A stripe assay will show whether inactive CEK7 axons grow towards RAGS cells and active CEK7 axons avoid RAGS axons.