Enhanced Antigen Retrieval of Amyloid β Immunohistochemistry: Re-evaluation of Amyloid β Pathology in Alzheimer Disease and Its Mouse Model
Enhanced Antigen Retrieval of Amyloid β Immunohistochemistry: Re-evaluation of Amyloid β Pathology in Alzheimer Disease and Its Mouse Model
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DOI：
10.1369/0022155412456379
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发表时间：
2012-10-01
影响因子：
3.2
通讯作者：
Kitamoto, Tetsuyuki
中科院分区：
文献类型：
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作者：
Kai, Hideaki;Shin, Ryong-Woon;Kitamoto, Tetsuyuki
老年斑是淀粉样β蛋白(Aβ)的细胞外沉积，是阿尔茨海默病(AD)的病理特征之一。作为Aβ沉积的标准免疫组织化学检测方法，抗Aβ免疫组织化学结合甲酸(FA)抗原修复(AR)已被广泛使用。在这里，我们提出了一种更有效的Aβ抗原AR。在AD及其小鼠模型的脑切片上，EDTA缓冲液中的高压灭菌加热或蛋白酶K消化加FA处理的双重组合增强了Aβ的免疫反应。进一步的三重组合：用蛋白酶K(P)消化，在EDTA缓冲液中高压灭菌加热(A)，以及FA处理(F)，当按顺序使用时，产生更强的免疫反应性。我们的PAF方法显著地显示了AD中各种形式的Aβ沉积，这些沉积以前没有被清楚地检测到，并在AD和小鼠模型中发现了许多微小尺寸的斑块。Aβ负荷的定量结果表明，PAF法的AR效应是FA法的1.86倍(在老年人脑中)和4.64倍(在小鼠脑中)。因此，PAF方法有可能成为迄今为止研究AD及其小鼠模型中Aβ病理的最灵敏的工具。(《组织化学细胞化学杂志》60：761-769,2012)
Senile plaques, extracellular deposits of amyloid beta peptide (A beta), are one of the pathological hallmarks of Alzheimer disease (AD). As the standard immunohistochemical detection method for A beta deposits, anti-A beta immunohistochemistry combined with antigen retrieval (AR) by formic acid (FA) has been generally used. Here, we present a more efficient AR for A beta antigen. On brain sections of AD and its mouse model, a double combination of either autoclave heating in EDTA buffer or digestion with proteinase K plus FA treatment reinforced A beta immunoreactivity. A further triple combination of digestion with proteinase K (P), autoclave heating in EDTA buffer (A), and FA treatment (F), when employed in this order, gave a more enhanced immunoreactivity. Our PAF method prominently visualized various forms of A beta deposits in AD that have not been clearly detected previously and revealed numerous minute-sized plaques both in AD and the mouse model. Quantification of A beta loads showed that the AR effect by the PAF method was 1.86-fold (in the aged human brain) and 4.64-fold (in the mouse brain) higher than that by the FA method. Thus, the PAF method could have the potential to be the most sensitive tool so far to study A beta pathology in AD and its mouse model. (J Histochem Cytochem 60:761-769, 2012)