A series of N-terminal epitope tagged Hdh knock-in alleles expressing normal and mutant huntingtin: their application to understanding the effect of increasing the length of normal Huntingtin's polyglutamine stretch on CAG140 mouse model pathogenesis.
A series of N-terminal epitope tagged Hdh knock-in alleles expressing normal and mutant huntingtin: their application to understanding the effect of increasing the length of normal Huntingtin's polyglutamine stretch on CAG140 mouse model pathogenesis.
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一系列的N末端表位标记了表达正常和突变体亨廷顿的HDH敲入等位基因：它们用于理解增加亨廷顿丁素多谷氨酰胺伸展长度对CAG140小鼠模型发病机理的效果的应用。
DOI：
10.1186/1756-6606-5-28
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发表时间：
2012-08-14
期刊：
影响因子：
3.6
通讯作者：
Zeitlin SO
中科院分区：
文献类型：
--
作者：
Zheng S;Ghitani N;Blackburn JS;Liu JP;Zeitlin SO
亨廷顿氏病（HD）是一种常染色体显性遗传的神经退行性疾病，其由亨廷顿蛋白（htt）内的多聚谷氨酰胺（polyQ）延伸的扩增引起，亨廷顿蛋白（htt）是HD基因的蛋白产物。虽然体外研究表明，突变体htt可以通过将野生型htt隔离到不溶性蛋白质聚集体中而以潜在的显性负性方式起作用，但正常htt polyQ延伸的长度和相邻的富含脯氨酸的区域（PRR）在调节HD小鼠模型发病机制中的作用目前尚不清楚。我们描述了一系列敲入HD小鼠模型的产生和表征，所述小鼠模型表达编码具有不同polyQ长度的N-末端血凝素（HA）或3xFlag表位标记的全长htt的小鼠HD基因（Hdh）的版本。（HA 7 Q-、3xFlag 7 Q-、3xFlag 20 Q-和3xFlag 140 Q-htt）和用人PRR取代相邻小鼠PRR（3xFlag 20 Q-和3xFlag 140 Q-htt）。使用免疫共沉淀和免疫组织化学分析，我们检测到可溶性全长正常7 Q-htt和突变体（140 Q）htt之间没有显著的相互作用，但我们确实观察到突变体htt聚集体中表位标记的正常htt的N末端片段。当编码正常小鼠htt的polyQ伸展和PRR的序列在也表达140 Q-htt的小鼠中被非致病性人序列替换时，纹状体内的聚集灶和htt包涵体的平均大小增加，沿着纹状体脂褐质和神经胶质增生的增加。在小鼠中，可溶性全长正常和突变htt主要是单体。在杂合基因敲入HD小鼠模型中，用正常人序列取代正常小鼠polyQ和PRR可加重某些神经病理表型。
Huntington’s disease (HD) is an autosomal dominant neurodegenerative disease that is caused by the expansion of a polyglutamine (polyQ) stretch within Huntingtin (htt), the protein product of the HD gene. Although studies in vitro have suggested that the mutant htt can act in a potentially dominant negative fashion by sequestering wild-type htt into insoluble protein aggregates, the role of the length of the normal htt polyQ stretch, and the adjacent proline-rich region (PRR) in modulating HD mouse model pathogenesis is currently unknown. We describe the generation and characterization of a series of knock-in HD mouse models that express versions of the mouse HD gene (Hdh) encoding N-terminal hemaglutinin (HA) or 3xFlag epitope tagged full-length htt with different polyQ lengths (HA7Q-, 3xFlag7Q-, 3xFlag20Q-, and 3xFlag140Q-htt) and substitution of the adjacent mouse PRR with the human PRR (3xFlag20Q- and 3xFlag140Q-htt). Using co-immunoprecipitation and immunohistochemistry analyses, we detect no significant interaction between soluble full-length normal 7Q- htt and mutant (140Q) htt, but we do observe N-terminal fragments of epitope-tagged normal htt in mutant htt aggregates. When the sequences encoding normal mouse htt’s polyQ stretch and PRR are replaced with non-pathogenic human sequence in mice also expressing 140Q-htt, aggregation foci within the striatum, and the mean size of htt inclusions are increased, along with an increase in striatal lipofuscin and gliosis. In mice, soluble full-length normal and mutant htt are predominantly monomeric. In heterozygous knock-in HD mouse models, substituting the normal mouse polyQ and PRR with normal human sequence can exacerbate some neuropathological phenotypes.
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影响因子：
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