Conformational templating of α-synuclein aggregates in neuronal-glial cultures.
Conformational templating of α-synuclein aggregates in neuronal-glial cultures.
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DOI：
10.1186/1750-1326-8-17
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发表时间：
2013-05-28
影响因子：
15.1
通讯作者：
Golde TE
中科院分区：
文献类型：
--
作者：
Sacino AN;Thomas MA;Ceballos-Diaz C;Cruz PE;Rosario AM;Lewis J;Giasson BI;Golde TE
遗传学研究已经确立了α-突触核蛋白（αS）在帕金森病（PD）中的致病作用，并且以路易体（LB）和路易神经突（LN）蛋白包涵体形式存在的αS聚集体定义了PD的病理特征。最近的研究表明，细胞外αS聚集可诱导细胞内αS病变，这支持了αS病变可通过“朊病毒样”自模板机制传播的假设。本研究通过在原代混合神经元-胶质细胞培养中使用重组野生型和pd -连锁突变体（A53T和E46K） αS进行播种，研究了αS细胞内聚集体构象模板化的可能性。我们发现，在混合原代培养的神经元和星形胶质细胞中，野生型和A53T αS原纤维以种子火焰状包裹体为主；而结构独特的E46K原纤维种子点状，圆形内含物。值得注意的是，这些培养中种子包涵体形成的差异反映了表达A53T或E46K αS突变体的转基因小鼠的包涵病理学差异。我们进一步表明，包合物的形态主要是由所施种子决定的，而不是αS表达的形式。我们也提供了初步证据表明αS包涵病理可以在原代星形胶质细胞培养中传代。这些研究首次证实了αS在培养细胞中可以通过形态自模板机制发生聚集。
Genetic studies have established a causative role for α-synuclein (αS) in Parkinson’s disease (PD), and the presence of αS aggregates in the form of Lewy body (LB) and Lewy neurite (LN) protein inclusions are defining pathological features of PD. Recent data has established that extracellular αS aggregates can induce intracellular αS pathologies supporting the hypothesis that αS pathology can spread via a “prion-like” self-templating mechanism. Here we investigated the potential for conformational templating of αS intracellular aggregates by seeding using recombinant wild-type and PD-linked mutant (A53T and E46K) αS in primary mixed neuronal-glial cultures. We find that wild-type and A53T αS fibrils predominantly seed flame-like inclusions in both neurons and astrocytes of mixed primary cultures; whereas the structurally distinct E46K fibrils seed punctate, rounded inclusions. Notably, these differences in seeded inclusion formation in these cultures reflect differences in inclusion pathology seen in transgenic mice expressing the A53T or E46K αS mutants. We further show that the inclusion morphology is dictated primarily by the seed applied rather than the form of αS expressed. We also provide initial evidence that αS inclusion pathology can be passaged in primary astrocyte cultures. These studies establish for the first time that αS aggregation in cultured cells can occur by a morphological self-templating mechanism.
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影响因子：
82.9
作者：
Conway, KA;Harper, JD;Lansbury, PT
通讯作者：
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作者：
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