Assessment of α-synuclein pathology:: A study of the BrainNet Europe consortium
Assessment of α-synuclein pathology:: A study of the BrainNet Europe consortium
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DOI：
10.1097/nen.0b013e3181633526
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发表时间：
2008-02-01
影响因子：
3.2
通讯作者：
Kretzschmar, Hans
中科院分区：
文献类型：
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作者：
Alafuzoff, Irina;Parkkinen, Laura;Kretzschmar, Hans
为了确定神经病理学家评估α-突触核蛋白免疫反应（α S-IR）结构的可靠性，来自BrainNet Europe 17个中心的28名评估人员使用组织微阵列（TMA）技术检查了α S-IR评估的当前方法和重现性。组织微阵列块由来自包含aS-IR结构的参与中心的样品构建。在每个中心对这些组织块的载玻片进行染色，并评估神经元核周包涵体、神经突和神经胶质细胞质包涵体。本研究分2个阶段进行。首先，用中心选择的抗体对TMA载玻片进行染色。在该阶段，59%的切片质量良好或可接受，使用的9种抗体中有4种表现一致。然而，α S-IR结构的解释和分类的差异导致实验室之间的结果不同。在第二阶段之前，神经病理学家参加了关于α S-IR结构评估的培训课程。根据第一阶段的结果，然后在第二阶段将使用指定抗原修复方法选择的抗体应用于TMA载玻片。当应用指定的染色和评价方法时，评价的所有26个随后染色的TMA切片均具有良好/可接受的质量，并且在评估是否存在特异性α S-IR结构方面实现了高度一致性。α S-IR神经元核周夹杂物的半定量评估产生的协议范围从49%到82%，最好的一致性在皮质核心样品。这些结果表明，严格的方法和二分法的评估（即确定是否存在α S-IR），应适用，半定量评估可以推荐仅用于皮质样本。此外，该研究表明，α S-IR结构的评分存在局限性。
To determine the reliability of assessment of alpha-synucleinimmunoreactive (alpha S-IR) structures by neuropathologists, 28 evaluators from 17 centers of BrainNet Europe examined current methods and reproducibility of alpha S-IR evaluation using a tissue microarray (TMA) technique. Tissue microarray blocks were constructed of samples from the participating centers that contained aS-IR structures. Slides from these blocks were stained in each center and assessed for neuronal perikaryal inclusions, neurites, and glial cytoplasmic inclusions. The study was performed in 2 phases. First, the TMA slides were stained with the antibody of the center's choice. In this phase, 59% of the sections were of good or acceptable quality, and 4 of 9 antibodies used performed consistently. Differences in interpretation and categorization of alpha S-IR structures, however, led to differing results between the laboratories. Prior to the second phase, the neuropathologists participated in a training session on the evaluation of alpha S-IR structures. Based on the results of the first phase, selected antibodies using designated antigen retrieval methods were then applied to TMA slides in the second phase. When the designated methods of both staining and evaluation were applied, all 26 subsequently stained TMA sections evaluated were of good/acceptable quality, and a high level of concordance in the assessment of the presence or absence of specific alpha S-IR structures was achieved. A semiquantitative assessment of alpha S-IR neuronal perikaryal inclusions yielded agreements ranging from 49% to 82%, with best concordance in cortical core samples. These results suggest that rigorous methodology and dichotomized assessment (i.e. determining the presence or absence of alpha S-IR) should be applied, and that semiquantitative assessment can be recommended only for the cortical samples. Moreover, the study demonstrates that there are limitations in the scoring of alpha S-IR structures.