Parkin localizes to the Lewy bodies of Parkinson disease and dementia with Lewy bodies
Parkin localizes to the Lewy bodies of Parkinson disease and dementia with Lewy bodies
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DOI：
10.1016/s0002-9440(10)61113-3
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发表时间：
2002-05-01
影响因子：
6
通讯作者：
Kosik, KS
中科院分区：
文献类型：
--
作者：
Schlossmacher, MG;Frosch, MP;Kosik, KS
α-突触核蛋白和parkin的突变会导致可遗传的帕金森病(PD)。我们假设神经元帕金，一种已知的E3泛素连接酶，促进路易小体(Lbs)的形成，这是帕金森病的病理标志。在这里，我们报告了在四种相关人类疾病的黑质切片中标记经典LBS的亲和纯化的Parkin抗体：散发性PD、遗传性Alphas连锁PD、LBS痴呆(DLB)和Lb阳性的Parkin相关PD。抗Parkin抗体还在DLB脑的内嗅皮层和扣带回皮质中检测到LBS，在散发性PD的交感神经节细胞中检测到Alphas包涵体。用共聚焦显微镜对DLB中脑切片进行双重标记显示，类似于90%的抗α反应LB也被针对氨基酸342到353的Parkin抗体检测到。相应地，Parkin蛋白，包括53-kd成熟异构体，存在于DLB皮质亲和力分离的LBS中。荧光、共振、能量转移和免疫电子显微镜显示AlpHas和parkin共同定位于脑干和皮质Lbs。在生化方面，Parkin似乎最丰富地存在于成年大鼠大脑的胞浆和突触后部分，但也存在于纯化的、富含Alpas的突触前元件中，这些元件还包含Parkin的E2结合伙伴UbcH7。我们的结论是，Parkin和UbcH7与Alphas一起存在于正常脑的亚细胞室中，并且Parkin经常与Alphas聚集体共存于PD和DLB的特征LB包涵体中。这些结果表明，在LB的形成过程中可能需要有功能的parkin蛋白。
Mutations in alpha-synuclein (alphaS) and parkin cause heritable forms of Parkinson disease (PD). We hypothesized that neuronal parkin, a known E3 ubiquitin ligase, facilitates the formation of Lewy bodies (LBs), a pathological hallmark of PD. Here, we report that affinity-purified parkin antibodies labeled classical LBs in substantia nigra sections from four related human disorders: sporadic PD, inherited alphaS-linked PD, dementia with LBs (DLB), and LB-positive, parkin-linked PD. Anti-parkin antibodies also detected LBs in entorhinal and cingulate cortices from DLB brain and alphaS inclusions in sympathetic gangliocytes from sporadic PD. Double labeling with confocal microscopy of DLB midbrain sections revealed that similar to90% of anti-alphaS-reactive LBs were also detected by a parkin antibody to amino acids 342 to 353. Accordingly, parkin proteins, including the 53-kd mature isoform, were present in affinity-isolated LBs from DLB cortex. Fluorescence resonance energy transfer and immunoelectron microscopy showed that alphaS and parkin co-localized within brainstem and cortical LBs. Biochemically, parkin appeared most enriched in cytosolic and postsynaptic fractions of adult rat brain, but also in purified, alphaS-rich presynaptic elements that additionally contained parkin's E2-binding partner, UbcH7. We conclude that parkin and UbcH7 are present with alphaS in subcellular compartments of normal brain and that parkin frequently co-localizes with alphaS aggregates in the characteristic LB inclusions of PD and DLB. These results suggest that functional parkin proteins may be required during LB formation.