Alzheimer's disease (AD) is characterized by toxic effects of amyloid β42 (Aβ42). Depending on the conditions, Aβ42 forms aggregates of distinct shapes and sizes. The most toxic are oligomers that bind to the cell membrane, where ganglioside GM1, sphingomyelin and cholesterol organized in lipid rafts are important interacting partners of Aβ42. Interaction of Aβ42 with clusters of GM1 induces shape changes and aggregation of amyloid, leading to cytotoxic effects.
The aim of the project is to monitor the effects of Aβ42 on endocytosis, lysosomal function, ROS production and other cellular processes in dependence on the lipid composition of the membrane. As a model system, the SH-SY5Y cell line and human neural stem cells (hNSCs) will be exploited. The use of hNSCs is a novelty in the study of AD which allows manipulating the composition of the membrane on cells with a neuronal phenotype. In particular, the effect of GM1 and cholesterol will be investigated. Due to the possible neuroprotective effect of docosahexaenoic acid, the impact of its action on Aβ42-induced toxicity will be analyzed. We also plan to use primary cultures from the mouse or rat hippocampus. For these investigations, we will use tissue culture techniques, different biochemical and proteomic analyses (SDS-PAGE, immunoblotting, determination of lipids), fluorescence microscopy, flow cytometry, centrifugation, etc.
Five relevant publications of the research group:
Rudajev V. and Novotny J. The Role of Lipid Environment in Ganglioside GM1-Induced Amyloid β Aggregation. Membranes (Basel). 2020 Sep 9;10(9):226.
Dolejší E, Liraz O, Rudajev V, Zimčík P, Doležal V and Michaelson DM. Apolipoprotein E4 reduces evoked hippocampal acetylcholine release in adult mice. J. Neurochem. 2016 Feb;136(3):503-9.
Janickova H, Rudajev V, Dolejsi E, Koivisto H, Jakubik J, Tanila H, El-Fakahany EE and Dolezal V. Lipid-Based Diets Improve Muscarinic Neurotransmission in the Hippocampus of Transgenic APPswe/PS1dE9 Mice. Curr. Alzheimer Res. 2015;12(10):923-31.
Janíčková H, Rudajev V, Zimčík P, Jakubík J, Tanila H, El-Fakahany EE and Doležal V. Uncoupling of M1 muscarinic receptor/G-protein interaction by amyloid β(1-42). Neuropharmacology. 2013 Apr;67:272-83.
Machová E, Rudajev V, Smycková H, Koivisto H, Tanila H and Dolezal V. Functional cholinergic damage develops with amyloid accumulation in young adult APPswe/PS1dE9 transgenic mice. Neurobiol. Dis. 2010 Apr;38(1):27-35.Apply to the project