Identification of synaptic accumulation of prion proten in Creutzfeldt-Jakob disease.

在2年的支持下，我们建立了水解高压灭菌预处理，以增强朊蛋白（PrP）的免疫组织化学。这一预处理为我们提供了异常的PrP在克雅氏病（CJD）患者中枢神经系统突触结构和CJD感染小鼠淋巴网状系统滤泡树突状细胞中积累的证据。为了证实PrP的突触积累，我们进行了生物化学和免疫组织化学检查。用抗PrP和抗synaptophysin双免疫标记显示PrP和synaptophysin的共定位。亚细胞分离中，突触体部分异常PrP浓度最高。1991年和1992年报告了这些结果。接下来，我们对不同的痴呆症患者进行了DNA测序。发现5个新的PrP基因突变。这些突变仅在CJD或Gerstmann-Straussler综合征中检测到。在这些突变中，我们可以根据异常PrP积累的分布将其分为斑块型和突触型。突触型积累型患者临床病程短，临床症状严重，而斑块型患者临床病程长，痴呆进展缓慢。因此，PrP的一级结构直接影响朊病毒疾病的表型。这些结果已经或将在以下参考文献中报告。

During 2 years' support, we established the hydrolytic autoclaving pretreatment to enhance the prion protein (PrP) immunohistochemistry. This pretreatment provided us that abnormal PrP accumulated in the synaptic structures in the central nervous system in patients with Creutzfeldt-Jakob disease (CJD), and in follicular dendritic cells in the lymphoreticular system in CJD-infected mice. To confirm synaptic accumulation of PrP, we examined biochemically and immuno histochemically. Double immunolabelling with anti-PrP and anti-synaptophysin revealed the colocalization of PrP and synaptophysin. In subcellular fractionation, synaptosomal fraction had the highest concentration of abnormal PrP. These results were reported in 1991 and 1992. Next, we did DNA sequencing from various demented persons. We found out novel 5 mutations of PrP gene. These mutations were detected only in CJD or Gerstmann-Straussler syndrome. Among these mutations, we can classify the plaque type or the synaptic type according to the distribution of abnormal PrP accumulation. Patients with synaptic type accumulation have a short clinical course with serious clinical signs, while patients with the plaque type have a long clinical course with slowly progressive dementia. Therefore, the primary structure of PrP directly influences the phenotype of prion diseases. These results were or will be reported in the following references.