Reiner Anton, Del Mar Nobel, Deng Yun-Ping, Meade Christopher A, Sun Zhiqiang, Goldowitz Daniel
Department of Anatomy & Neurobiology, College of Medicine, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
J Comp Neurol. 2007 Dec 20;505(6):603-29. doi: 10.1002/cne.21515.
The R6/2 mouse possesses mutant exon 1 of human Hdh, and R6/2 mice with 150 CAG repeats show neurological abnormalities by 10 weeks and die by 15 weeks. Few brain abnormalities, however, are evident at death, other than widespread ubiquitinated neuronal intranuclear inclusions (NIIs). We constructed R6/2t+/t- <--> wildtype (WT) chimeric mice to prolong survival of R6/2 cells and determine if neuronal death and/or neuronal injury become evident with longer survival. ROSA26 mice (which bear a lacZ transgene) were used as WT to distinguish between R6/2 and WT neurons. Chimeric mice consisting partly of R6/2 cells lived longer than pure R6/2 mice (up to 10 months), with the survival proportional to the R6/2 contribution. Genotypically R6/2 cells formed NIIs in the chimeras, and these NIIs grew only slightly larger than in 12-week pure R6/2 mice, even after 10 months. Additionally, neuropil aggregates formed near R6/2 neurons in chimeric mice older than 15 weeks. Thus, R6/2 neurons could survive well beyond 15 weeks in chimeras. Moreover, little neuronal degeneration was evident in either cortex or striatum by routine histological stains. Nonetheless, striatal shrinkage and ventricular enlargement occurred, and striatal projection neuron markers characteristically reduced in Huntington's disease were diminished. Consistent with such abnormalities, cortex and striatum in chimeras showed increased astrocytic glial fibrillary acidic protein. These results suggest that while cortical and striatal neurons can survive nearly a year with nuclear and extranuclear aggregates of mutant huntingtin, such lengthy survival does reveal cortical and striatal abnormality brought on by the truncated mutant protein.
R6/2小鼠携带人Hdh基因的突变外显子1,具有150个CAG重复序列的R6/2小鼠在10周时出现神经学异常,并在15周时死亡。然而,除了广泛存在的泛素化神经元核内包涵体(NIIs)之外,死亡时几乎没有明显的脑部异常。我们构建了R6/2t+/t- <--> 野生型(WT)嵌合小鼠,以延长R6/2细胞的存活时间,并确定随着存活时间延长,神经元死亡和/或神经元损伤是否会变得明显。ROSA26小鼠(携带lacZ转基因)用作野生型,以区分R6/2和野生型神经元。部分由R6/2细胞组成的嵌合小鼠比纯R6/2小鼠存活时间更长(长达10个月),其存活时间与R6/2细胞的占比成正比。从基因型上看,R6/2细胞在嵌合体中形成了NIIs,即使在10个月后,这些NIIs的大小也仅比12周龄的纯R6/2小鼠略大一点。此外,在超过15周龄的嵌合小鼠中,R6/2神经元附近形成了神经纤维聚集物。因此,R6/2神经元在嵌合体中可以存活超过15周。此外,通过常规组织学染色,在皮质或纹状体中几乎没有明显的神经元变性。尽管如此,纹状体萎缩和脑室扩大仍会发生,并且亨廷顿病中特征性减少的纹状体投射神经元标志物也会减少。与这些异常情况一致,嵌合体中的皮质和纹状体显示星形胶质细胞的胶质纤维酸性蛋白增加。这些结果表明,虽然皮质和纹状体神经元可以携带突变型亨廷顿蛋白的核内和核外聚集体存活近一年,但如此长的存活时间确实揭示了截短的突变蛋白所导致的皮质和纹状体异常。