Objective: To investigate the neurotoxic properties of alpha-synuclein (α-Syn) aggregation in different buffers including PBS, SBF and Tris on altered cell lines, SHSY5Y, SK-N-MC and PC12. These buffers are routinely used in α-Syn fibrillation researches which imitating natural buffers. Also, all applied cells have been widely employed in studies regarding Parkinson’s disease (PD).

Background: The aggregation of α-Syn, a presynaptic protein present at high concentrations in neural cells of the central nervous system, has been demonstrated to play a key role in patients suffering from PD. In addition to PD, α-Syn aggregation is the cytopathological symbol of some other diseases known as synucleinopathies. These aggregations have the ability to transmit form cell-to-cell and are believed to have a critical role in neurodegeneration or can act as a secondary pattern in neural cell death.

Methods: Fibrillation of recombinant human α-Syn was carried out in individual buffers at a variety of concentrations for 24 h, and fibril formation was then assessed by ThT fluorescence intensity, fluorescence microscopy images, AFM images and Congo red absorbance. MTT, LDH and ROS measurements were then used to evaluate the cytotoxicity of the different α-Syn aggregates formed on the discrete noted cell lines.

Results: Results indicated that individual buffers and different concentrations of α-Syn have diverse effects on the rate of accumulation and aggregation of the protein. In addition, such aggregations have a different toxic influence on the three different cell lines.

Conclusions: These results indicate that precautions should be taken with regard to the type of buffer and most importantly the cell type, when carrying out studies that focus on mechanism or evaluate the effects of various pharmaceutical agents. The importance of this finding will become more evident when a drug is entered into clinical trials. Prior to this stage, it is the cell culture method that is important in assessing the effect of the drug. These results together highlight the effect of different buffers and cell lines in studying α-Syn role in PD that should be considered in preclinical investigations.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/diverse-effects-of-alpha-synuclein-fibrils-on-various-neuron-cell-lines-using-different-buffers/