Maturation dependent neuroprotection against alcohol

成熟依赖的酒精神经保护作用

基本信息

批准号：
7140310
负责人：
BAHRI KARACAY
金额：
$ 20.7万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-05 至 2008-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Fetal alcohol syndrome is an important cause of mental retardation. Exposure of the developing brain to alcohol can induce neuronal death, which contributes strongly to the learning deficits and neurological problems associated with FAS. Understanding the factors that influence neuronal vulnerability to alcohol-induced cell death in the developing brain is of considerable importance. We hypothesize that this maturation-dependent alcohol resistance is due to the acquisition of an inherent neuroprotective signaling pathway mediated by VPAC1, a G-protein-coupled receptor. This proposal will elucidate the function and molecular mechanisms of the VPAC1 signaling pathway, including one of its key downstream effector molecules, cAMP-Responsive Element Binding protein (CREB), in protecting developing neurons against alcohol-induced death. The first specific aim will assess the protective effect of the VPAC1 signaling pathway against alcohol toxicity in vitro and in vivo. We will trace the developmental expression of the pathway components and their downstream target neuroprotective genes and will determine whether the ontogeny of these genes coincides with the acquisition of time-dependent alcohol resistance. We will compare the seventy of alcohol-induced neuronal loss in Purkinje and granule cell cultures derived from mice with and without a functional VPAC1 receptor and will determine whether neurons lacking this receptor can be "rescued' from alcohol toxicity by the targeted delivery of VPAC1 carried by gene therapy vectors. We will determine whether the ectopic expression of VPAC 1 during the vulnerable period can prevent alcohol-induced death in cerebellar neurons. The second specific aim will explore the molecular mechanism by which the VPAC1 signaling pathway leads to neuronal protection. We will selectively silence VPAC1 and CREB using siRNA and shRNA technology and will assess the effect of this gene silencing on neuronal survival. We will determine whether specific silencing of VPAC1 or CREB can abolish the resistance of cerebellar granule and Purkinje neurons against alcohol-induced cell death during the resistant phase. We will determine whether silencing of VPAC1 or CREB affects the expression of neuroprotective and anti-apoptolic genes carrying Camp response elements in their promoters. These studies will provide vital information regarding the role and mechanism of an important G-protein-mediated signaling pathway in neuroprotection against alcohol. The reagents and results derived from these in vitro experiments will lay the foundation for the gene therapy studies planned for in vivo applications in the future.

描述（由申请人提供）：胎儿酒精综合征是智力低下的重要原因。发育中的大脑暴露于酒精会诱导神经元死亡，这对与FAS相关的学习缺陷和神经系统问题有很大贡献。了解影响神经元脆弱性在发育中的大脑中对酒精引起的细胞死亡的因素至关重要。我们假设这种成熟依赖性的酒精耐药性是由于获得了由G蛋白偶联受体VPAC1介导的固有神经保护信号通路。该建议将阐明VPAC1信号通路的功能和分子机制，包括其关键的下游效应分子之一，cAMP响应元件结合蛋白（CREB），在保护发育神经元免受酒精诱导的死亡中的发育中。第一个具体目的将评估VPAC1信号通路在体外和体内对酒精毒性的保护作用。我们将追踪途径成分及其下游靶神经保护基因的发育表达，并将确定这些基因的个体发育是否与获得时间依赖性酒精耐药性的获得相吻合。我们将比较伯爵和颗粒细胞培养物的七十个葡萄酒诱导的神经元丧失，这些细胞和颗粒细胞培养有或没有功能性VPAC1受体的小鼠，并将确定是否可以通过基因疗法在vlulapy dulapy vulapy vector中携带的VPAC递送的vpac1 cander canse vpace vectiC inder-nife vpac inder-nife vpac inder vecor-nife vecor-nife vpac inder vecopic inder-nife vpac1 cance dy vpac inder vpac infor vpac1 cyciic inder。我们是否可以从酒精毒性中“拯救”。小脑神经元的死亡。在抗性阶段，对酒精诱导的细胞死亡的神经元和CREB的沉默是否会影响启动子中cAMP反应元件的神经保护元素和抗寄生虫反应元件的表达。这些研究将提供有关重要的G蛋白介导的信号传导途径在针对酒精的神经保护中的作用和机制的重要信息。这些体外实验得出的试剂和结果将为未来计划用于体内应用的基因治疗研究奠定基础。