CEK7 RAGS RECEPTOR LIGAND RELATIONSHIP IN RETINA AND TEC

这个建议调查了一个假定的排斥因子，RAGS， 内源性鸡顶盖蛋白被认为参与了 视网膜神经节细胞在发育过程中的指导作用。在轴突 Drescher已经证明，轴突可以避免生长成 RAGS标记细胞。候选人提议测试 RAGS的内源性受体是CEK 7，一种EPH受体酪氨酸 在鸡视网膜中显示激酶。实验结果很好- 有明确的概念基础。这些技术很容易 申办方可获得所有必要的试剂， 创造基于与其他物种的同源性， 该建议将创建非常有用的开发系统， 来研究轴突引导演示RAGS/CEK 7功能 相互作用将增加我们对抑制因子的知识 其阻止移植后视神经功能再生 神经损伤 该项目分为两个具体目标。首先，候选人 建议证明配体和 受体，并确定哪些受体亚型可以结合 破布。将使用RAGS和CEK 7的重组来源。激活 将通过自磷酸化来测量受体的活性。第二 目的是研究RAGS/CEK 7相互作用是否实际上是 用小鸡视网膜神经节细胞做实验以显示 由于排斥相互作用的特异性，CEK 7将被 发色团辅助激光灭活。CEK 7周转率 将确定激光灭活后的视网膜功能， 轴突引导可以在存在或不存在下研究轴突生长。 活性CEK 7。条纹分析将显示无活性CEK 7轴突是否 向RAGS细胞生长，并且活性CEK 7轴突避开RAGS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.