Prion Pathogenesis Is Faithfully Reproduced in Cerebellar Organotypic Slice Cultures
Prion Pathogenesis Is Faithfully Reproduced in Cerebellar Organotypic Slice Cultures
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DOI：
10.1371/journal.ppat.1002985
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发表时间：
2012-11-01
期刊：
影响因子：
6.7
通讯作者：
Aguzzi, Adriano
中科院分区：
文献类型：
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作者：
Falsig, Jeppe;Sonati, Tiziana;Aguzzi, Adriano
朊病毒在体内引起神经变性，但朊病毒感染的培养细胞不显示细胞毒性。这阻碍了朊病毒诱导神经变性的机制研究。在这里，我们报告说，朊病毒感染的培养器官型小脑切片（COCS）经历了进行性海绵状神经变性密切复制朊病毒疾病，三种不同的朊病毒株引起三种不同的朊病毒蛋白沉积模式。当PrP从神经元中遗传去除时，神经变性不会发生，并且全面的药理学筛选表明，已知拮抗朊病毒复制的化合物可以消除神经变性。朊病毒感染的COCS和小鼠导致增强的胞衬蛋白裂解，表明钙蛋白酶或半胱天冬酶的发病机制的参与。因此，神经毒性和胞衬蛋白裂解被钙蛋白酶抑制剂阻止，但不能被半胱天冬酶抑制剂阻止，而朊病毒复制则不受阻碍。因此钙蛋白酶抑制剂可以解除朊病毒复制与其神经毒性后遗症的耦合。这些数据验证了COCS作为一个强大的模型系统，忠实地再现了朊病毒感染的大多数形态学标志。COCS对药理学操作的精致可及性有助于认识钙蛋白酶在神经毒性中的作用，并显着扩展了严格解剖朊病毒发病机制所需的工具。
Prions cause neurodegeneration in vivo, yet prion-infected cultured cells do not show cytotoxicity. This has hampered mechanistic studies of prion-induced neurodegeneration. Here we report that prion-infected cultured organotypic cerebellar slices (COCS) experienced progressive spongiform neurodegeneration closely reproducing prion disease, with three different prion strains giving rise to three distinct patterns of prion protein deposition. Neurodegeneration did not occur when PrP was genetically removed from neurons, and a comprehensive pharmacological screen indicated that neurodegeneration was abrogated by compounds known to antagonize prion replication. Prion infection of COCS and mice led to enhanced fodrin cleavage, suggesting the involvement of calpains or caspases in pathogenesis. Accordingly, neurotoxicity and fodrin cleavage were prevented by calpain inhibitors but not by caspase inhibitors, whereas prion replication proceeded unimpeded. Hence calpain inhibition can uncouple prion replication from its neurotoxic sequelae. These data validate COCS as a powerful model system that faithfully reproduces most morphological hallmarks of prion infections. The exquisite accessibility of COCS to pharmacological manipulations was instrumental in recognizing the role of calpains in neurotoxicity, and significantly extends the collection of tools necessary for rigorously dissecting prion pathogenesis.