Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease.
Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease.
复制标题
从R6/2的大脑和亨廷顿氏病小鼠模型中捕获的相同的低聚物和原纤维结构。
DOI：
10.1093/hmg/ddp467
复制
发表时间：
2010-01-01
影响因子：
3.5
通讯作者：
Bates GP
中科院分区：
文献类型：
--
作者：
Sathasivam K;Lane A;Legleiter J;Warley A;Woodman B;Finkbeiner S;Paganetti P;Muchowski PJ;Wilson S;Bates GP
亨廷顿病(HD)是一种起病晚的神经退行性疾病，其神经病理特征是神经纤维团和核包涵体的存在。然而，HD患者或HD小鼠模型大脑中存在的聚集体结构的轮廓以及特定聚集体结构在疾病发病机制中的相对贡献尚不清楚。我们已经使用Seprion配体建立了一种基于酶联免疫吸附试验(ELISA)的高灵敏方法，用于定量HD小鼠模型组织中聚集的聚谷氨酰胺。结合电子显微镜、原子力显微镜(AFM)和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)对配体分离的聚集体结构进行了研究。我们发现，从R6/2和HdhQ150敲入小鼠的脑中提取的低聚物、原纤维和纤维聚集体非常相似。使用AFM，我们确定从两个小鼠模型的脑中分离出的纳米球形低聚物的尺寸与从重组Huntingtin外显子1蛋白产生的尺寸相同。最后，检测外显子1 Htt表位的抗体可以不同地识别SDS-PAGE凝胶上的配基捕获物质。Seprion-Ligand ELISA提供了一种具有良好统计能力的分析方法，可用于临床前药效治疗试验或评估潜在治疗靶点的遗传操作对总负荷的影响。这一点，加上识别HD小鼠组织中聚集物种的光谱的能力，将有助于我们理解这些结构如何与HD的发病机制相关，以及它们的形成是否可以被操纵以获得治疗益处。
Huntington's disease (HD) is a late-onset neurodegenerative disorder that is characterized neuropathologically by the presence of neuropil aggregates and nuclear inclusions. However, the profile of aggregate structures that are present in the brains of HD patients or of HD mouse models and the relative contribution of specific aggregate structures to disease pathogenesis is unknown. We have used the Seprion ligand to develop a highly sensitive enzyme-linked immunosorbent assay (ELISA)-based method for quantifying aggregated polyglutamine in tissues from HD mouse models. We used a combination of electron microscopy, atomic force microscopy (AFM) and sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) to investigate the aggregate structures isolated by the ligand. We found that the oligomeric, proto-fibrillar and fibrillar aggregates extracted from the brains of R6/2 and HdhQ150 knock-in mice were remarkably similar. Using AFM, we determined that the nanometre globular oligomers isolated from the brains of both mouse models have dimensions identical to those generated from recombinant huntingtin exon 1 proteins. Finally, antibodies that detect exon 1 Htt epitopes differentially recognize the ligand-captured material on SDS–PAGE gels. The Seprion-ligand ELISA provides an assay with good statistical power for use in preclinical pharmacodynamic therapeutic trials or to assess the effects of the genetic manipulation of potential therapeutic targets on aggregate load. This, together with the ability to identify a spectrum of aggregate species in HD mouse tissues, will contribute to our understanding of how these structures relate to the pathogenesis of HD and whether their formation can be manipulated for therapeutic benefit.
登录
查看更多内容
影响因子：
5.3
作者：
Díaz-Hernández, M;Moreno-Herrero, F;Lucas, JJ
通讯作者：
Lucas, JJ
影响因子：
16.8
作者：
Li, Pingwei;Huey-Tubman, Kathryn E.;Bjorkman, Pamela J.
通讯作者：
Bjorkman, Pamela J.
影响因子：
3.8
作者：
Ko, J;Ou, S;Patterson, PH
通讯作者：
Patterson, PH
影响因子：
4.8
作者：
Legleiter, Justin;Lotz, Gregor P.;Muchowski, Paul J.
通讯作者：
Muchowski, Paul J.
影响因子：
2.9
作者：
Chromy, BA;Nowak, RJ;Klein, WL
通讯作者：
Klein, WL