A novel, high-efficiency cellular model of fibrillar alpha-synuclein inclusions and the examination of mutations that inhibit amyloid formation.
A novel, high-efficiency cellular model of fibrillar alpha-synuclein inclusions and the examination of mutations that inhibit amyloid formation.
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DOI：
10.1111/j.1471-4159.2010.06592.x
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发表时间：
2010-04
影响因子：
4.7
通讯作者：
Giasson BI
中科院分区：
文献类型：
--
作者：
Waxman EA;Giasson BI
胞浆内α-突触核蛋白(α-SYN)淀粉样包涵体是帕金森病、路易体病痴呆和多系统萎缩的主要病理特征。这些聚集体的形成和抑制所涉及的机制是深入研究的领域。本研究描述了一种用于研究α-SYN聚集的新的细胞模型，该模型结合了成核依赖的聚集和磷酸钙沉淀的新函数。培养的细胞很容易被诱导形成大的、细胞质的α-SYN丝状聚集体，这些聚集体被过度磷酸化，通常是泛素化的，硫代黄素阳性。这些细胞聚集体在大多数转基因细胞中形成，并招募了大约一半内源性表达的α-SYN。使用这个系统，我们在体外检测了抑制α-SYN淀粉样蛋白形成的单点突变。三个突变(V66P、T72P和T75P)显著阻碍了该细胞模型中α-SYN的聚集。T75P突变体可以在体外消除野生型α-SYN的淀粉样蛋白形成，但不能阻止野生型α-SYN的细胞聚集体。这些研究表明，α-SYN形成细胞聚集体的倾向可能比单独的体外研究更明显。这种新的高效的α-SYN聚集的细胞模型是一个有价值的系统，可以用来进一步了解α-SYN聚集并为未来的治疗方法的产生创造条件。
Intracytoplasmic alpha-synuclein (α-syn) amyloidogenic inclusions are a major pathological feature of Parkinson's disease, dementia with Lewy body disease, and multiple systems atrophy. The mechanisms involved in the formation and inhibition of these aggregates are areas of intense investigation. The present study characterizes a novel cellular model for the study of α-syn aggregation, incorporating nucleation-dependent aggregation and a new function for calcium phosphate precipitation. Cultured cells were readily induced to develop large, cytoplasmic α-syn filamentous aggregates that were hyperphosphorylated, often ubiquitinated and thioflavin positive. These cellular aggregates formed in the majority of transfected cells and recruited approximately half of endogenously expressed α-syn. Using this system, we examined single-point mutations that inhibit α-syn amyloid formation in vitro. Three mutations (V66P, T72P, and T75P) significantly hindered α-syn aggregation in this cell model. The T75P mutant, which could abrogate amyloid formation of wild-type α-syn in vitro, did not prevent wild-type α-syn cellular aggregates. These studies suggest that the propensity of α-syn to form cellular aggregates may be more pronounced than in isolated in vitro studies. This novel high-efficiency cellular model of α-syn aggregation is a valuable system that may be used to further understand α-syn aggregation and allow for the generation of future therapeutics.
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影响因子：
82.9
作者：
Conway, KA;Harper, JD;Lansbury, PT
通讯作者：
Lansbury, PT
DOI：
10.1016/j.bbrc.2008.01.140
发表时间：
2008-04-11
影响因子：
3.1
作者：
Koo, Hyun-Jung;Lee, Hak-Joo;Im, Hana
通讯作者：
Im, Hana
DOI：
10.1083/jcb.143.7.1883
发表时间：
1998-12-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Johnston JA;Ward CL;Kopito RR
通讯作者：
Kopito RR
影响因子：
2.9
作者：
Conway, KA;Harper, JD;Lansbury, PT
通讯作者：
Lansbury, PT
影响因子：
4.7
作者：
Ahn, KJ;Paik, SR;Kim, J
通讯作者：
Kim, J