Sequestration of free cholesterol in cell membranes by prions correlates with cytoplasmic phospholipase A(2) activation

Bate, C; Tayebi, M; Williams, A

Abstract

Background: The transmissible spongiform encephalopathies (TSEs), otherwise known as the prion diseases, occur following the conversion of the normal cellular prion protein ( PrPC) to an alternatively folded isoform ( PrPSc). The accumulation of PrPSc within the brain leads to neurodegeneration through an unidentified mechanism. Since many neurodegenerative disorders including prion, Parkinson's and Alzheimer's diseases may be modified by cholesterol synthesis inhibitors, the effects of prion infection on the cholesterol balance within neuronal cells were examined. Results: We report the novel observation that prion infection altered the membrane composition and significantly increased total cholesterol levels in two neuronal cell lines (ScGTI and ScN2a cells). There was a significant correlation between the concentration of free cholesterol in ScGTI cells and the amounts of PrPSc. This increase was entirely a result of increased amounts of free cholesterol, as prion infection reduced the amounts of cholesterol esters in cells. These effects were reproduced in primary cortical neurons by the addition of partially purified PrPSc, but not by PrPC. Crucially, the effects of prion infection were not a result of increased cholesterol synthesis. Stimulating cholesterol synthesis via the addition of mevalonate, or adding exogenous cholesterol, had the opposite effect to prion infection on the cholesterol balance. It did not affect the amounts of free cholesterol within neurons; rather, it significantly increased the amounts of cholesterol esters. Immunoprecipitation studies have shown that cytoplasmic phospholipase A(2) (cPLA(2)) co-precipitated with PrPSc in ScGTI cells. Furthermore, prion infection greatly increased both the phosphorylation of cPLA(2) and prostaglandin E-2 production. Conclusion: Prion infection, or the addition of PrPSc, increased the free cholesterol content of cells, a process that could not be replicated by the stimulation of cholesterol synthesis. The presence of PrPSc increased solubilisation of free cholesterol in cell membranes and affected their function. It increased activation of the PLA(2) pathway, previously implicated in PrPSc formation and in PrPSc-mediated neurotoxicity. These observations suggest that the neuropathogenesis of prion diseases results from PrPSc altering cholesterol-sensitive processes. Furthermore, they raise the possibility that disturbances in membrane cholesterol are major triggering events in neurodegenerative diseases.

Citation

Bate, C., Tayebi, M., & Williams, A. Sequestration of free cholesterol in cell membranes by prions correlates with cytoplasmic phospholipase A(2) activation. BMC Biology, 6, https://doi.org/10.1186/1741-7007-6-8

• Journal Article Type: Article
• Deposit Date: Nov 11, 2014
• Journal: BMC Biology
• Electronic ISSN: 1741-7007
• Publisher: BioMed Central
• Volume: 6
• DOI: https://doi.org/10.1186/1741-7007-6-8
• Public URL: https://rvc-repository.worktribe.com/output/1428506