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PrP fragment 106-126 is toxic to cerebral endothelial cells expressing PrP(C)
M A Deli1, S Sakaguchi, R Nakaoke
1Department of Pharmacology 1, School of Medicine, Nagasaki University, Japan.
Abstract:
A hydrophobic, fibrillogenic peptide fragment of human prion protein (PrP106-126) had in vitro toxicity to neurons expressing cellular prion protein (PrP(C)). In this study, we proved that primary cultures of mouse cerebral endothelial cells (MCEC) express PrP(C). Incubation of MCEC with PrP106-126 (25-200 microM) caused a dose-dependent toxicity assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase release, bis-benzimide staining for nuclear morphology, and trypan blue exclusion test. Pentosan polysulphate (50-100 microg/ml), a drug effective in scrapie prophylaxis, dose-dependently attenuated the injury. MCEC cultures from mice homogenous for the disrupted PrP gene were resistant to the toxicity of PrP106-126. In conclusion, cerebral endothelium expressing PrP(C) may be directly damaged during spongiform encephalopathies.
Insights
Prion protein fragment PrP106-126 damages mouse brain endothelial cells expressing cellular prion protein (PrP(C)). This damage, relevant to spongiform encephalopathies, was reduced by pentosan polysulphate.
Area of Science:
- Neuroscience
- Cell Biology
- Prion Disease Research
Background:
- Prion diseases involve abnormal prion protein formation.
- Cellular prion protein (PrP(C)) is implicated in neuroprotection and disease.
- The role of cerebral endothelium in prion disease pathogenesis is unclear.
Purpose of the Study:
- To investigate the expression of PrP(C) in mouse cerebral endothelial cells (MCEC).
- To determine the toxicity of a prion protein fragment (PrP106-126) to MCEC.
- To evaluate the protective effect of pentosan polysulphate against PrP106-126-induced MCEC toxicity.
Main Methods:
- Primary MCEC cultures were established and characterized for PrP(C) expression.
- MCEC were exposed to varying concentrations of PrP106-126.
- Cell viability was assessed using MTT assay, LDH release, nuclear morphology (bis-benzimide), and trypan blue exclusion.
- The effect of pentosan polysulphate was evaluated on MCEC exposed to PrP106-126.
- MCEC from PrP gene-knockout mice were used as a control.
Main Results:
- MCEC were confirmed to express PrP(C).
- PrP106-126 induced a dose-dependent toxicity in MCEC.
- Pentosan polysulphate significantly attenuated PrP106-126-induced MCEC injury.
- MCEC from PrP-deficient mice were resistant to PrP106-126 toxicity.
Conclusions:
- Cerebral endothelial cells express PrP(C) and are susceptible to damage by PrP106-126.
- This suggests that the cerebral endothelium may be a direct target during spongiform encephalopathies.
- Pentosan polysulphate demonstrates potential therapeutic benefit by protecting endothelial cells.