LRRK2-mediated molecular and synaptic events in the striatum

项目摘要 帕金森病(Parkinson‘s Disease，PD)是第二常见的神经退行性疾病。基因突变 LRRK2与遗传性和散发性帕金森病有关。LRRK2富含刺状物质 背侧纹状体投射神经元(SPN)。然而，LRRK2在SPN中的生理作用仍然存在 难以捉摸。我们之前的观察表明致病的R1441C突变和异常之间存在联系 SPN中的PKA信号。然而，分析受到以下固有分辨率的限制 常规的生化方法和显微镜检查。在这项提案中，我们寻求提供更准确的 关于SPN中LRRK2突变的病理生理后果的信息。特别是，它是我们的 中心假设LRRK2R1441C突变导致SPN中的多巴胺能信号异常。由此产生的 纹状体功能障碍反过来又导致帕金森病的症状。为了实现这一目标，我们将研究 LRRK2R1441C突变改变多巴胺信号、皮质纹状体传递、皮质纹状体可塑性和 通过PKA信号异常调节SPN的兴奋性。此外，纹状体依赖运动的评估 结合药物操作的学习将提供对LRRK2R1441C突变的影响的洞察 在一个完整的动物环境中。结合标准的细胞、分子和电生理学 方法，我们的调查将利用尖端方法的组合，克服 到目前为止阻碍进展的障碍。这些包括，LRRK2突变小鼠，纹状体通路特异性 (CRE和记者)小鼠、病毒基因传递和超分辨率成像。成功地实现了 这些目标将极大地促进我们对帕金森病背后的机制的理解，在这样做的过程中， 推动未来帕金森病患者新疗法的开发。

Project Summary Parkinson's disease (PD) is the second most common neurodegenerative disease of aging. Mutations in LRRK2 are associated with both inherited and sporadic forms of PD. LRRK2 is highly enriched in spiny projection neurons (SPN) in the dorsal striatum. However, the physiological role of LRRK2 in SPNs remains elusive. Our previous observations suggest a linkage between the pathogenic R1441C mutation and aberrant PKA signaling in SPNs. However, the analyses were limited by the inherent resolution provided by conventional biochemical approaches and microscopy. In this proposal, we seek to provide more precise information about the pathophysiological consequences of LRRK2 mutations in SPNs. In particular, it is our central hypothesis that LRRK2R1441C mutation leads to aberrant dopaminergic signaling in SPNs. The resultant striatal dysfunction, in turn, contributes to the symptomatology of PD. To pursue this, we will examine if LRRK2R1441C mutation alters dopamine signaling, corticostriatal transmission, corticostriatal plasticity, and excitability in SPNs through PKA signaling dysregulation. Moreover, assessment of striatal-dependent motor learning along with pharmacological manipulations will provide insight on the effects of LRRK2R1441C mutation in a whole-animal setting. In conjunction with standard cellular, molecular, and electrophysiological approaches, our investigations will capitalize on a combination of cutting-edge approaches that overcome obstacles that have impeded progress to date. These include, LRRK2 mutant mice, striatal pathway-specific (Cre and reporter) mice, viral gene delivery, and super-resolution imaging. The successful achievement of these aims will significantly advance our understanding of the mechanisms underlying PD, and in doing so, will promote the development of new therapies for PD patients in the future.