Deposition pattern and subcellular distribution of disease-associated prion protein in cerebellar organotypic slice cultures infected with scrapie
Deposition pattern and subcellular distribution of disease-associated prion protein in cerebellar organotypic slice cultures infected with scrapie
复制标题
感染瘙痒病的小脑器官切片培养物中疾病相关朊病毒蛋白的沉积模式和亚细胞分布
作者：
Hanna Wolf;André Hossinger;Andrea Fehlinger;Sven Büttner;Valerie L. Sim;Debbie McKenzie;I. Vorberg
器官型小脑切片代表了一个合适的模型，在复杂的细胞环境中表征和操纵朊病毒复制。器官型切片概括了朊病毒的病理学，并且在没有血脑屏障的情况下适用于药物测试。到目前为止，疾病特异性朊蛋白在器官型切片中的细胞和亚细胞分布尚不清楚。在这里，我们报告的同时检测疾病特异性朊蛋白和中枢神经系统标志物在野生型小鼠小脑切片感染小鼠适应朊病毒株22 L。切片中的疾病特异性朊病毒蛋白分布与体内分布非常相似，主要在分子层和浦肯野细胞层中显示颗粒状斑点样沉积。双重免疫染色发现异常朊病毒蛋白在神经胶质细胞和相关的神经元，星形胶质细胞和小胶质细胞，但在浦肯野细胞的情况。建立的协议，同时免疫组织化学检测疾病特异性朊病毒蛋白和细胞标志物，使详细分析朊病毒复制和药物疗效的体外模型的中枢神经系统。
Organotypic cerebellar slices represent a suitable model for characterizing and manipulating prion replication in complex cell environments. Organotypic slices recapitulate prion pathology and are amenable to drug testing in the absence of a blood-brain-barrier. So far, the cellular and subcellular distribution of disease-specific prion protein in organotypic slices is unclear. Here we report the simultaneous detection of disease-specific prion protein and central nervous system markers in wild-type mouse cerebellar slices infected with mouse-adapted prion strain 22L. The disease-specific prion protein distribution profile in slices closely resembles that in vivo, demonstrating granular spot like deposition predominately in the molecular and Purkinje cell layers. Double immunostaining identified abnormal prion protein in the neuropil and associated with neurons, astrocytes and microglia, but absence in Purkinje cells. The established protocol for the simultaneous immunohistochemical detection of disease-specific prion protein and cellular markers enables detailed analysis of prion replication and drug efficacy in an ex vivo model of the central nervous system.