Comparison of mHTT Antibodies in Huntington's Disease Mouse Models Reveal Specific Binding Profiles and Steady-State Ubiquitin Levels with Disease Development.
Comparison of mHTT Antibodies in Huntington's Disease Mouse Models Reveal Specific Binding Profiles and Steady-State Ubiquitin Levels with Disease Development.
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DOI：
10.1371/journal.pone.0155834
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Brooks SP
中科院分区：
文献类型：
--
作者：
Bayram-Weston Z;Jones L;Dunnett SB;Brooks SP
亨廷顿氏病（HD）的细胞病理特征是突变的亨廷顿蛋白（mHTT）聚集到包涵体中。本文比较了5种常用的mHTT抗体（S830、MW8、EM48、1C2、ubiquitin）对5种常用HD小鼠模型（R6/1、YAC128、HdhQ92、B6 HdhQ150、B6 x129/Ola HdhQ150）在2个年龄的敏感性，确定了各模型最敏感的抗体；mHTT抗体结合是否因聚集阶段不同而不同（弥漫性或直接包涵）；泛素作为泛素蛋白体系统在聚集过程中的作用与疾病的发展有关。即使模型仅受背景菌株（HdhQ150）的影响，也显示出抗体结合的独特特征。MW8在所有细胞系中检测直率的内含物高度敏感，而EM48、泛素和1C2在所有模型中表现出一致的染色，而与年龄或mHTT形式无关。其中MW8和S830最敏感，1C2最不敏感。无论年龄大小，每个模型的泛素水平都是稳定的。泛素在年轻的YAC128小鼠中特别敏感，直到12个月大的时候才显示出包涵体的缺失，这表明对弥漫性mHTT有很高的亲和力。数据表明，关于聚合量化的模型的概括可能是无效的，mHTT抗体结合是小鼠模型所特有的，对包涵体发育的变化很敏感。
Huntington’s disease (HD) cellular pathology is characterised by the aggregation of mutant huntingtin (mHTT) protein into inclusion bodies. The present paper compared the sensitivity of five widely used mHTT antibodies (S830; MW8; EM48; 1C2; ubiquitin) against mice from five commonly used HD mouse models (R6/1; YAC128; HdhQ92; B6 HdhQ150; B6 x129/Ola HdhQ150) at two ages to determine: the most sensitive antibodies for each model; whether mHTT antibody binding differed depending on aggregation stage (diffuse versus frank inclusion); the role of ubiquitin during aggregation as the ubiquitin proteosome system has been implicated in disease development. The models demonstrated unique profiles of antibody binding even when the models varied only by background strain (HdhQ150). MW8 was highly sensitive for detecting frank inclusions in all lines whereas EM48, ubiquitin and 1C2 demonstrated consistent staining in all models irrespective of age or form of mHTT. MW8 and S830 were the most sensitive antibodies with 1C2 the least. Ubiquitin levels were stable for each model regardless of age. Ubiquitin was particularly sensitive in young YAC128 mice that demonstrate an absence of inclusions until ~12 months of age suggesting high affinity to mHTT in its diffuse form. The data indicate that generalisations across models regarding the quantification of aggregations may not be valid and that mHTT antibody binding is unique to the mouse model and sensitive to changes in inclusion development.