Grad Leslie I, Pokrishevsky Edward, Cashman Neil R
Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, F137, 2211 Wesbrook Mall, Vancouver, BC, Canada, V6T 285.
Methods Mol Biol. 2017;1658:357-367. doi: 10.1007/978-1-4939-7244-9_24.
The prion hypothesis has extended to the fatal motor neuron disease, amyotrophic lateral sclerosis (ALS), as a means to explain the spatiotemporal spread of pathology from one or more focal points through the neuroaxis. About 20% of inheritable cases of ALS are due to mutation in the gene encoding the Cu/Zn superoxide dismutase (SOD1), causing the protein to misfold and form neurotoxic aggregates. Mutant SOD1 has been shown to impart its misfold onto natively folded wild-type SOD1 in living cells. Furthermore, misfolded wild-type SOD1 can itself induce further rounds of propagated SOD1 misfolding. Finally, this prion-like mechanism of propagated SOD1 misfolding can be transmitted from cell to cell in human cell culture. Here, we describe a protocol for the induction of wild-type SOD1 misfolding inside living cells and its subsequent transmission from cell to cell in a prion-like fashion.
朊病毒假说已扩展至致命性运动神经元疾病——肌萎缩侧索硬化症（ALS），作为一种解释病理学从一个或多个病灶点通过神经轴进行时空传播的方式。约20%的可遗传ALS病例是由于编码铜/锌超氧化物歧化酶（SOD1）的基因突变，导致该蛋白质错误折叠并形成神经毒性聚集体。已证明突变型SOD1会在活细胞中将其错误折叠传递给天然折叠的野生型SOD1。此外，错误折叠的野生型SOD1本身可诱导进一步轮次的SOD1错误折叠传播。最后，这种SOD1错误折叠传播的朊病毒样机制可在人类细胞培养中在细胞间传递。在此，我们描述了一种在活细胞内诱导野生型SOD1错误折叠及其随后以朊病毒样方式在细胞间传递的方案。
