Immunohistochemical detection of the delayed formation of non-fibrillar large amyloid-β aggregates.
Immunohistochemical detection of the delayed formation of non-fibrillar large amyloid-β aggregates.
复制标题
免疫组织化学检测非纤维状大淀粉样蛋白-β 聚集体的延迟形成。
DOI：
10.1111/gtc.12332
复制
发表时间：
2016
期刊：
影响因子：
2.1
通讯作者：
Yoshimune K
中科院分区：
文献类型：
--
作者：
Ochiishi T;Itakura A;Liu L;Akatsu H;Kohno H;Nishimura M;Yoshimune K
由淀粉样β蛋白（Aβ）组成的老年斑的发生是阿尔茨海默病（AD）的主要神经病理学标志。我们先前开发并表征了单克隆抗体31 - 2和75 - 2，它们分别特异性结合直径大于220和50 nm的非纤维状Aβ1- 42聚集体。在此，我们报告了使用这些抗体来检测外源性Aβ1- 42在培养的大鼠海马神经元中的聚集。Aβ1-42转染后6 ~ 24 h，抗体75 - 2免疫标记几乎所有转染的神经元，而31 - 2阳性细胞仅限于部分转染的神经元，数量逐渐增加。F19 S/L34 P ‐突变体Aβ1-42的表达显示聚集趋势较低，导致对两种抗体的免疫反应性明显降低。我们还对AD患者的颞叶皮质进行了化学研究，发现31 - 2优先标记了大淀粉样斑块亚群的核心。发现31 - 2-免疫反应性斑块的相对数量与神经元缠结的Braak分期相关，但与淀粉样斑块无关。这些结果表明，在AD脑的培养神经元和淀粉样斑块中，31 - 2-反应性Aβ聚集体的形成具有延迟的时程。
The occurrence of senile plaques consisting of amyloid‐β protein (Aβ) is a major neuropathological hallmark of Alzheimer's disease (AD). We previously developed and characterized monoclonal antibodies 31‐2 and 75‐2 that specifically bind to nonfibrillar Aβ1–42aggregates with diameters of more than 220 and 50 nm, respectively. Here, we report the use of these antibodies to examine the aggregation of exogenous Aβ1–42in cultured rat hippocampal neurons. From 6 to 24 h after transfection of Aβ1–42, antibody 75‐2 immunolabeled almost all transfected neurons, whereas 31‐2‐positive cells were restricted to a part of the transfected neurons and gradually increased in number. Expression of the F19S/L34P‐mutant Aβ1–42, which showed less of a tendency to aggregate, resulted in clearly reduced immunoreactivity to both antibodies. We also immunohistochemically investigated the temporal cortices of patients with AD and found that 31‐2 preferentially labeled the cores of a subpopulation of large amyloid plaques. The relative number of 31‐2‐immunoreactive plaques was found to correlate with the Braak stages of neurofibrillary tangles, but not with that of amyloid plaques. These results suggest that 31‐2‐reactive Aβ aggregates develop with a delayed time course in cultured neurons and amyloid plaques of AD brains.