α-Synuclein Seeding Assay Using RT-QuIC.
α-Synuclein Seeding Assay Using RT-QuIC.
复制标题
使用 RT-QuIC 进行 α-突触核蛋白接种测定。
DOI：
10.1007/978-1-0716-1495-2_1
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Hattori Nobutaka
中科院分区：
文献类型：
--
作者：
Okuzumi Ayami;Hatano Taku;Fukuhara Takeshi;Ueno Shinichi;Nukina Nobuyuki;Imai Yuzuru;Hattori Nobutaka
突触核蛋白病是与α-突触核蛋白（αSyn）错误折叠和聚集相关的神经退行性疾病。它们包括帕金森病、路易体痴呆和多系统萎缩。在每种疾病中，已经提出αSyn的聚集体代表αSyn的不同构象菌株，导致自繁殖并在细胞间传播。α - syn聚集体是通过种子聚合机制生长的。在此之前，已经通过实时震动诱导转化（RT-QuIC）实验揭示了朊病毒蛋白聚集过程中种子转化的机制。通过加入荧光染料硫黄素-t进一步改进，可以实时监测动力学变化，并对极低水平的种子聚集体进行高灵敏度检测。在诊断应用中，据报道，αSyn RT-QuIC的特异性在82%到100%之间，而其敏感性在70%到100%之间，这是基于在多个不同实验室进行的一项研究。此外，αSyn RT-QuIC方法可用于研究不同αSyn菌株在突触核蛋白病中的生化特性。在本文中，我们描述了αSyn RT-QuIC检测的详细协议。
Synucleinopathies are neurodegenerative diseases that are associated with the misfolding and aggregation of α-synuclein (αSyn). They include Parkinson’s disease, dementia with Lewy bodies, and multiple system atrophy. In each disease, it has been proposed that aggregates of αSyn represent different conformational strains of αSyn, leading to self-propagation and spreading from cell to cell. It has been considered that αSyn aggregates grow by seeded polymerization mechanisms. Previously, the mechanism of seed conversion in prion protein aggregation has been exploited by real-time quaking-induced conversion (RT-QuIC) assay. It was further refined by incorporating the fluorescent dye thioflavin-T, which enabled the real-time monitoring of kinetic changes with a highly sensitive detection of seed aggregates present at an extremely low level. In an application for diagnostics, it has been reported that αSyn RT-QuIC exhibits specificity between 82% and 100%, while its sensitivity varies between 70% and 100%, on the basis of a study in which this assay was performed at multiple different laboratories. Furthermore, it has been suggested that the αSyn RT-QuIC method can be applied to study the biochemical characteristics of different αSyn strains among synucleinopathies. In this article, we describe the detailed protocols for αSyn RT-QuIC assays.
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DOI：
10.1007/978-1-4939-8820-4_2
发表时间：
2019-01-01
期刊：
PROTEIN MISFOLDING DISEASES
影响因子：
--
作者：
Saijo, Eri;Groveman, Bradley R.;Caughey, Byron
通讯作者：
Caughey, Byron
影响因子：
6.7
作者：
Wilham JM;Orrú CD;Bessen RA;Atarashi R;Sano K;Race B;Meade-White KD;Taubner LM;Timmes A;Caughey B
通讯作者：
Caughey B
朊病毒疾病的生物学和遗传学。
影响因子：
10.5
作者：
S. Prusiner
通讯作者：
S. Prusiner
DOI：
10.1016/s0368-1742(61)80013-1
发表时间：
1961-01-01
期刊：
JOURNAL OF COMPARATIVE PATHOLOGY AND THERAPEUTICS
影响因子：
--
作者：
PATTISON, IH;MILLSON, GC
通讯作者：
MILLSON, GC