CNTF RECEPTOR ALPHA REGULATION AND FUNCTION

神经退行性疾病由多种有毒物质引起， 导致退化和死亡的创伤性和遗传性侮辱 神经细胞的。与之形成鲜明对比的是，成年脊髓运动神经元 不仅完全存活下来的创伤性损伤(轴突切断术) 其他神经元群体的不可逆转的损失，其轴突功能 让他们的目标恢复神经。更好地理解大体上 未知的、细胞和分子机制导致了这种情况 令人印象深刻的伤病反应显然有很大的潜力 在设计有效的治疗方法的斗争中受益 神经退行性疾病。我们假设，和几个独立的 一系列间接证据表明，CNTF受体a(CNTFRa) 对成体的生存和再生做出了重要贡献 脊神经切断后的脊髓运动神经元。然而，迄今为止的研究 尚未直接确定：1)地点、时间和程度 CNTFRa蛋白在这种损伤后表达，因此 潜在活性(特定目标1)；2)CNTFRa信号 转导事件是由病变诱导的(特定目标2)；以及3) (S)CNTFRa在动物的生存和再生中扮演什么角色 损伤神经元(特定目标3)。我们将使用坐骨神经损伤 模型和免疫组织化学定位损伤诱导的大鼠脑内病变 CNTFRa在亚细胞水平的表达 CNTFRa抗体，并通过原位证实我们的发现 杂交，Northern和Western blotting(特定目标1)。五花八门 病变诱导的“候选”CNTFRa信号转导事件 以免疫组织化学、原位杂交或 逆行运输程序，并被确认为CNTFRa依赖 通过功能阻断抑制体内CNTFRA功能 抗体、充当拮抗剂的突变CNTF和反义 DNA(特异性靶标2)。CNTFRa在生存、神经递质中的作用 损伤后的表型和轴突再生也将同样 通过测量以下因素对这些属性的影响确定 体内CNTFRa功能的抑制(特定目标3)。除了……之外 极大地扩展了目前对CNTFRa在 脊髓运动神经元的存活和再生，建议 实验将构成对成人的第一次直接活体检查 CNTFRa函数。

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.