Characterization of HTT Inclusion Size, Location, and Timing in the zQ175 Mouse Model of Huntington's Disease: An In Vivo High-Content Imaging Study
Characterization of HTT Inclusion Size, Location, and Timing in the zQ175 Mouse Model of Huntington's Disease: An In Vivo High-Content Imaging Study
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DOI：
10.1371/journal.pone.0123527
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发表时间：
2015-04-10
期刊：
影响因子：
3.7
通讯作者：
Kwak, Seung
中科院分区：
文献类型：
--
作者：
Carty, Nikisha;Berson, Nadege;Kwak, Seung
亨廷顿病（Huntington's disease，HD）是一种由亨廷顿基因CAG三核苷酸重复扩增引起的常染色体显性遗传性神经退行性疾病。HD的主要病理学标志包括突变亨廷顿蛋白（mHTT）的内含物，主要在尾状核中的神经元的损失，以及多个脑区域的萎缩。然而，早期序列的组织学事件，表现在区域和细胞特异性的方式还没有得到很好的表征。在这里，我们使用高含量的组织学方法来精确地监测HTT表达的变化，并在最近表征的zQ 175基因敲入小鼠系中表征mHTT蛋白包涵体的沉积动力学。我们对2-12月龄小鼠组织切片中的单个皮质和纹状体细胞进行了自动化多参数定量分析，并证实了该模型中mHTT内含物年龄相关增加的生化报告。我们还发现了不同的区域和次区域的动态包含数量，大小和分布与亚细胞分辨率。我们使用zQ 175小鼠纹状体中总HTT的病毒介导的抑制作为治疗相关但异质性转导策略的一个例子，以证明该平台在细胞基础上定量评估靶点参与和结果的成功应用。
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the huntingtin gene. Major pathological hallmarks of HD include inclusions of mutant huntingtin (mHTT) protein, loss of neurons predominantly in the caudate nucleus, and atrophy of multiple brain regions. However, the early sequence of histological events that manifest in region-and cell-specific manner has not been well characterized. Here we use a high-content histological approach to precisely monitor changes in HTT expression and characterize deposition dynamics of mHTT protein inclusion bodies in the recently characterized zQ175 knock-in mouse line. We carried out an automated multi-parameter quantitative analysis of individual cortical and striatal cells in tissue slices from mice aged 2-12 months and confirmed biochemical reports of an age-associated increase in mHTT inclusions in this model. We also found distinct regional and subregional dynamics for inclusion number, size and distribution with subcellular resolution. We used viral-mediated suppression of total HTT in the striatum of zQ175 mice as an example of a therapeutically-relevant but heterogeneously transducing strategy to demonstrate successful application of this platform to quantitatively assess target engagement and outcome on a cellular basis.