Viral expression of ALS-linked ubiquilin-2 mutants causes inclusion pathology and behavioral deficits in mice.
Viral expression of ALS-linked ubiquilin-2 mutants causes inclusion pathology and behavioral deficits in mice.
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DOI：
10.1186/s13024-015-0026-7
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发表时间：
2015-07-08
影响因子：
15.1
通讯作者：
McFarland NR
中科院分区：
文献类型：
--
作者：
Ceballos-Diaz C;Rosario AM;Park HJ;Chakrabarty P;Sacino A;Cruz PE;Siemienski Z;Lara N;Moran C;Ravelo N;Golde TE;McFarland NR
UBQLN 2突变最近与家族性肌萎缩侧索硬化症（ALS）和ALS痴呆相关。UBQLN 2编码泛素-2，其是泛素样蛋白家族的成员，其促进将泛素化蛋白递送至蛋白酶体进行降解。为了研究ubiquilin-2在ALS中的潜在作用，我们使用重组腺相关病毒（rAAV）载体在原代神经胶质细胞培养物和发育中的新生小鼠大脑中表达UBQLN 2和三种已鉴定的ALS相关突变体（P497 H，P497 S和P506 T）。在原代培养中，rAAV 2/8介导的UBQLN 2突变体的表达导致包涵体和不溶性聚集体。在出生后第0天用表达野生型或突变型UBQLN 2的rAAV 2/8对FVB小鼠进行脑室内注射，导致脑中广泛、持续的泛素-2表达。与野生型相比，突变体UBQLN 2表达诱导了显著的病理学变化，在周围神经元中具有大的神经元、细胞质内含物和泛素-2阳性聚集体。泛素-2包涵体与泛素、p62/SQSTM、视神经磷酸酶共定位，偶尔与TDP-43共定位，但α-突触核蛋白、神经丝、tau和FUS阴性。突变体UBLQN 2表达也导致硫磺素-S阳性包涵体/聚集体。表达突变形式UBQLN 2 β的小鼠在3-4个月时发展出运动表型，包括非特异性抱握和旋转杆缺陷。这些发现表明，UBQLN 2突变体（P497 H，P497 S和P506 T）诱导蛋白质病并导致行为缺陷，支持“毒性”功能获得，这可能有助于ALS病理学。这些数据还证实，我们的rAAV模型可用于快速评估各种UBQLN 2突变的病理后果，并提供了一个灵活的系统，以进一步询问泛素在神经变性中的分子机制。
UBQLN2 mutations have recently been associated with familial forms of amyotrophic lateral sclerosis (ALS) and ALS-dementia. UBQLN2 encodes for ubiquilin-2, a member of the ubiquitin-like protein family which facilitates delivery of ubiquitinated proteins to the proteasome for degradation. To study the potential role of ubiquilin-2 in ALS, we used recombinant adeno-associated viral (rAAV) vectors to express UBQLN2 and three of the identified ALS-linked mutants (P497H, P497S, and P506T) in primary neuroglial cultures and in developing neonatal mouse brains. In primary cultures rAAV2/8-mediated expression of UBQLN2 mutants resulted in inclusion bodies and insoluble aggregates. Intracerebroventricular injection of FVB mice at post-natal day 0 with rAAV2/8 expressing wild type or mutant UBQLN2 resulted in widespread, sustained expression of ubiquilin-2 in brain. In contrast to wild type, mutant UBQLN2 expression induced significant pathology with large neuronal, cytoplasmic inclusions and ubiquilin-2-positive aggregates in surrounding neuropil. Ubiquilin-2 inclusions co-localized with ubiquitin, p62/SQSTM, optineurin, and occasionally TDP-43, but were negative for α-synuclein, neurofilament, tau, and FUS. Mutant UBLQN2 expression also resulted in Thioflavin-S-positive inclusions/aggregates. Mice expressing mutant forms of UBQLN2 variably developed a motor phenotype at 3–4 months, including nonspecific clasping and rotarod deficits. These findings demonstrate that UBQLN2 mutants (P497H, P497S, and P506T) induce proteinopathy and cause behavioral deficits, supporting a “toxic” gain-of-function, which may contribute to ALS pathology. These data establish also that our rAAV model can be used to rapidly assess the pathological consequences of various UBQLN2 mutations and provides an agile system to further interrogate the molecular mechanisms of ubiquilins in neurodegeneration.
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影响因子：
15.1
作者：
Sacino AN;Thomas MA;Ceballos-Diaz C;Cruz PE;Rosario AM;Lewis J;Giasson BI;Golde TE
通讯作者：
Golde TE
影响因子：
12.7
作者：
Wu Q;Liu M;Huang C;Liu X;Huang B;Li N;Zhou H;Xia XG
通讯作者：
Xia XG
DOI：
10.1016/s0167-4889(01)00164-1
发表时间：
2002-01-30
影响因子：
5.1
作者：
Wu, SL;Mikhailov, A;Avruch, J
通讯作者：
Avruch, J
DOI：
10.1016/j.bbrc.2008.02.086
发表时间：
2008-05-02
影响因子：
3.1
作者：
Kim, Tae-Yeon;Kim, Eunmin;Yoon, Jong-Bok
通讯作者：
Yoon, Jong-Bok
影响因子：
3.5
作者：
Rothenberg, Cara;Srinivasan, Deepa;Monteiro, Mervyn J.
通讯作者：
Monteiro, Mervyn J.