Role of LRRK2 in idiopathic Parkinson's disease

LRRK 2突变导致家族性帕金森病（PD），在某些人群中，可能占所有帕金森病的40%。 案例3. LRRK 2基因座也包含“特发性”PD（iPD）的风险因子;然而，LRRK 2在PD中的作用是不确定的。 典型的iPD不清楚。此外，LRRK 2与其他与PD相关的基因和蛋白质的关系，如 α-突触核蛋白也相对未被探索。虽然突变型LRRK 2引起神经变性的机制 尚不完全确定，但一般认为致病突变可能与激酶 活性，至少在原位在完整的细胞。然而，理解内源性野生型 iPD中的LRRK 2缺乏实用的高分辨率测定其活化状态。 我们已经开发并验证了一对具有出色解剖分辨率的新型邻位连接测定法， 快速提供关于LRRK 2的激活状态、细胞定位和生理调节剂的信息。的 测定基于（i）S1292磷酸化和（ii）14-3-3从LRRK 2解离。使用这种和其他分析，我们 有初步证据表明：（i）LRRK 2在iPD的黑质纹状体神经元中被激活;（ii）亚致死浓度的LRRK 2在iPD的黑质纹状体神经元中被激活。 鱼藤酮激活LRRK 2;（iii）体内α-突触核蛋白的过表达激活黑质纹状体神经元中的LRRK 2;（iv） LRRK 2的激活是由氧化机制介导的。我们现在建议研究LRRK 2在iPD中的作用及其在iPD中的作用。 与α-突触核蛋白和线粒体的潜在生理相互作用。我们将研究以下问题：（1） 人iPD脑组织中LRRK 2的激活状态是什么？(2)内源性LRRK 2激活在多大程度上 在鱼藤酮鼠身上做模型(3)是的！α-突触核蛋白激活内源性WT LRRK 2？(4)氧化应激会激活 LRRK2？

Mutations in LRRK2 cause familial Parkinson disease (PD) and, in some populations, may account for up to 40% of all cases3. The LRRK2 gene locus also contains a risk factor for `idiopathic' PD (iPD); however, the role of LRRK2 in typical iPD is not clear. Furthermore, the relationship of LRRK2 to other genes and proteins associated with PD, such as α-synuclein, is also relatively unexplored. While the mechanism(s) by which mutant LRRK2 causes neurodegeneration are not entirely certain, it is generally believed that disease-causing mutations may be associated with increased kinase activity, at least in situ in intact cells. However, a critical barrier to understanding the role of endogenous, wildtype LRRK2 in iPD is the absence of a practical, high-resolution assay for its activation state. We have developed and validated a pair of novel proximity ligation assays with excellent anatomical resolution that can rapidly provide information regarding activation state, cellular localization and physiological regulators of LRRK2. The assay is based on (i) S1292 phosphorylation and (ii) dissociation of 14-3-3 from LRRK2. Using this and other assays, we have preliminary evidence that (i) LRRK2 is activated in nigrostriatal neurons in iPD; (ii) sublethal concentrations of rotenone activate LRRK2; (iii) overexpression of α-synuclein in vivo activates LRRK2 in nigrostriatal neurons; (iv) LRRK2 activation is mediated by oxidative mechanisms. We now propose to examine the role of LRRK2 in iPD and its potential physiological interactions with α-synuclein and mitochondria. We will investigate the following questions: (1) What is the activation state of LRRK2 in human iPD brain tissue? (2) To what extent can endogenous LRRK2 activation be modeled in the rotenone rat? (3) Does!α-synuclein activate endogenous WT LRRK2? (4) Does oxidative stress activate LRRK2?