| First Author | Woerman AL | Year | 2015 |
| Journal | Proc Natl Acad Sci U S A | Volume | 112 |
| Issue | 35 | Pages | E4949-58 |
| PubMed ID | 26286986 | Mgi Jnum | J:226705 |
| Mgi Id | MGI:5698305 | Doi | 10.1073/pnas.1513426112 |
| Citation | Woerman AL, et al. (2015) Propagation of prions causing synucleinopathies in cultured cells. Proc Natl Acad Sci U S A 112(35):E4949-58 |
| abstractText | Increasingly, evidence argues that many neurodegenerative diseases, including progressive supranuclear palsy (PSP), are caused by prions, which are alternatively folded proteins undergoing self-propagation. In earlier studies, PSP prions were detected by infecting human embryonic kidney (HEK) cells expressing a tau fragment [TauRD(LM)] fused to yellow fluorescent protein (YFP). Here, we report on an improved bioassay using selective precipitation of tau prions from human PSP brain homogenates before infection of the HEK cells. Tau prions were measured by counting the number of cells with TauRD(LM)-YFP aggregates using confocal fluorescence microscopy. In parallel studies, we fused alpha-synuclein to YFP to bioassay alpha-synuclein prions in the brains of patients who died of multiple system atrophy (MSA). Previously, MSA prion detection required approximately 120 d for transmission into transgenic mice, whereas our cultured cell assay needed only 4 d. Variation in MSA prion levels in four different brain regions from three patients provided evidence for three different MSA prion strains. Attempts to demonstrate alpha-synuclein prions in brain homogenates from Parkinson's disease patients were unsuccessful, identifying an important biological difference between the two synucleinopathies. Partial purification of tau and alpha-synuclein prions facilitated measuring the levels of these protein pathogens in human brains. Our studies should facilitate investigations of the pathogenesis of both tau and alpha-synuclein prion disorders as well as help decipher the basic biology of those prions that attack the CNS. |
