School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, People's Republic of China.
J Mol Neurosci. 2013 Nov;51(3):734-48. doi: 10.1007/s12031-013-0057-x. Epub 2013 Jul 6.
αB-crystallin is a member of the small heat shock protein family constitutively presenting in brains at a relatively low level. To address the alteration of αB-crystallin in prion disease, the αB-crystallin levels in the brains of scrapie agent 263 K-infected hamsters were analyzed. The levels of αB-crystallin were remarkably increased in the brains of 263 K-infected hamsters, showing a time-dependent manner along with incubation time. Immunohistochemical (IHC) and immunofluorescent (IFA) assays illustrated more αB-crystallin-positive signals in the regions of the cortex and thalamus containing severe astrogliosis. Double-stained IFA verified that the αB-crystallin signals colocalized with the enlarged glial fibrillary acidic protein-positive astrocytes, but not with neuronal nuclei-positive cells. IHC and IFA of the serial brain sections of infected hamsters showed no colocalization and correlation between PrP(Sc) deposits and αB-crystallin increase. Moreover, increased αB-crystallin deposits were observed in the brain sections of parietal lobe of a sporadic Creutzfeldt-Jakob disease (sCJD) case, parietal lobe and thalamus of a G114V genetic CJD case, and thalamus of a fatal family insomnia (FFI) case, but not in a parietal lobe of FFI where only very mild astrogliosis was addressed. Additionally, the molecular interaction between αB-crystallin and PrP was only observed in the reactions of recombinant proteins purified from Escherichia coli, but not either in that of brain homogenates or in that of the cultured cell lysates expressing human PrP and αB-crystallin. Our data indicate that brain αB-crystallin is abnormally upregulated in various prion diseases, which is coincidental with astrogliosis. Direct interaction between αB-crystallin and PrP seems not to be essential during the pathogenesis of prion infection.
αB-晶体蛋白是小热休克蛋白家族的成员,在大脑中以相对较低的水平组成型存在。为了研究朊病毒病中αB-晶体蛋白的变化,分析了感染 263 K 传染性剂的仓鼠脑中的αB-晶体蛋白水平。在感染 263 K 的仓鼠脑中,αB-晶体蛋白水平显着增加,并且随着孵育时间呈时间依赖性。免疫组织化学(IHC)和免疫荧光(IFA)检测表明,在含有严重星形胶质增生的皮质和丘脑区域中,αB-晶体蛋白阳性信号更多。双染 IFA 验证了αB-晶体蛋白信号与放大的神经胶质纤维酸性蛋白阳性星形胶质细胞共定位,但与神经元核阳性细胞不共定位。感染仓鼠的连续脑切片的 IHC 和 IFA 显示,PrP(Sc)沉积物与αB-晶体蛋白增加之间没有共定位和相关性。此外,在散发性克雅氏病(sCJD)病例的顶叶脑切片、G114V 遗传 CJD 病例的顶叶和丘脑以及致命家族性失眠症(FFI)病例的丘脑中观察到增加的αB-晶体蛋白沉积物,但在仅轻度星形胶质增生的 FFI 顶叶中没有观察到。此外,仅在从大肠杆菌中纯化的重组蛋白反应中观察到αB-晶体蛋白和 PrP 之间的分子相互作用,而在脑匀浆或表达人 PrP 和αB-晶体蛋白的培养细胞裂解物的反应中均未观察到。我们的数据表明,在各种朊病毒病中,脑αB-晶体蛋白异常上调,这与星形胶质增生一致。在朊病毒感染的发病机制中,αB-晶体蛋白与 PrP 之间的直接相互作用似乎不是必需的。