CNTF RECEPTOR ALPHA REGULATION AND FUNCTION

神经退行性疾病是由多种毒性， 导致退化和死亡的创伤和遗传损伤 的神经元。 与此形成鲜明对比的是，成年脊髓运动神经元群体 不仅完全存活创伤性损伤（轴突切断术）， 在其他神经元群体中，它们的轴突功能上 重新激活他们的目标 更好地理解大部分 未知的，细胞和分子机制负责这一点 令人印象深刻的伤害反应显然将具有很大的潜力 在设计有效治疗方法的斗争中受益， 神经退行性疾病 我们假设，几个独立的 一系列间接证据表明，CNTF受体a（CNTFRa） 对成虫的存活和再生有着至关重要的作用 脊髓运动神经元轴突切断术。 然而，迄今为止的研究表明， 我没有直接确定：1）在哪里，何时以及在多大程度上 CNTFRa蛋白在这样的损伤后表达，并且因此被称为 潜在活性（具体目标1）; 2）CNTFRa信号 转导事件由损伤诱导（特异性目标2）;以及3） CNTFRa在植物的存活和再生中起什么作用？ 损伤神经元（具体目标3）。 我们将利用坐骨神经损伤 模型和免疫化学地图病变引起的变化， CNTFRa表达在亚细胞水平的分辨率与我们的抗- CNTFRa抗体，并通过原位杂交证实了我们的发现。 杂交、北方和西方印迹（特异性目的1）。 各种 损伤诱导的“候选”CNTFRa信号转导事件将被 用免疫组织化学、原位杂交或 逆行转运程序，并确定为CNTFRa依赖性 通过功能阻断抑制体内CNTFRA功能 抗体，作为拮抗剂的突变CNTF，和反义 DNA（特定目标2）。 CNTFRa在生存中的作用，神经递质 表型和损伤后的轴突再生类似地将 通过测量这些属性如何受到 抑制体内CNTFR α功能（具体目标3）。 除了 大大扩展了目前对CNTFRa在以下方面作用的理解： 脊髓运动神经元的存活和再生， 实验将构成第一个直接在体内检查成人 CNTFRa功能。

Neurodegenerative disorders result from a wide variety of toxic, traumatic and genetic insults which lead to the degeneration and death of neurons. In sharp contrast, adult spinal motor neuron populations not only completely survive traumatic injury (axotomy) that produces irreversible loss in other neuronal populations, their axons functionally reinnervate their targets. A better understanding of the largely unknown, cellular and molecular mechanisms responsible for this impressive injury response obviously would be of great potential benefit in the struggle to design effective treatments for neurodegenerative disorders. We hypothesize, and several independent lines of indirect evidence suggest, that CNTF receptor a (CNTFRa) makes a critical contribution to the survival and regeneration of adult spinal motor neurons following axotomy. However, studies to date have not directly determined: 1) where, when and to what extent CNTFRa protein is expressed following such a lesion and is therefore potentially active (Specific Aim 1); 2) what CNTFRa signal transduction events are induced by the lesion (Specific Aim 2); and 3) what role(s) CNTFRa plays in the survival and regeneration of the injured neurons (Specific Aim 3). We will use the sciatic nerve lesion model and immunohistochemically map lesion-induced changes in CNTFRa expression at a subcellular level of resolution with our anti- CNTFRa antibodies and confirm our findings through in situ hybridization, northern and western blots (Specific Aim 1). Various lesion-induced, "candidate" CNTFRa signal transduction events will be characterized with immunohistochemistry, in situ hybridization or retrograde transport procedures and identified as CNTFRa dependent by inhibition of in vivo CNTFRA function with function blocking antibodies, a mutant CNTF that acts as an antagonist, and antisense DNA (Specific Aim 2). CNTFRa's role in survival, neurotransmitter phenotype and axonal regeneration following lesion similarly will be determined by measuring how these properties are influenced by inhibition of in vivo CNTFRa function (Specific aim 3). In addition to greatly expanding the current understanding of CNTFRa's role in spinal motor neuron survival and regeneration, the proposed experiments will constitute the first direct in vivo examination of adult CNTFRa function.