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Information × Registration Number 0217U002344, 0115U004200 , R & D reports Title Modeling of molecular processes underlying Parkinson's disease in yeast cells and analyzing the impact of exogenous and endogenous factors on the processes of aggregation and degradation of human alpha-synuclein popup.stage_title Head Stasyk Oleh Volodymyrovych, Кандидат біологічних наук Registration Date 06-02-2017 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 For modeling processes inherent for neurons at Parkinson's disease, pGLG61-SNCA vector for expression of the genetic construct SNCA-GFP consisting of gene sequence of human alpha-synuclein (SNCA), fused ithw gene of green fluorescent protein (yEGFP), in Hansenula polymorpha yeast cells was constructed. Since the pathology of dopaminergic neurons of the substantia nigra in the midbrain at Parkinson's disease often develops on the background of overexpression of alpha-synuclein, the multi-copy integrants selected for resistance to Geneticin, and showed increased GFP fluorescence-labeled alpha-synuclein in the cytosol were used in our work. It is found that in a medium with 1% glucose (physiological yeast culture conditions), the proportion of dead cells of the strain NCYC 495 / SNCA was significantly higher compared to the wild type strain, indicating toxicity of heterologous alpha-synuclein gene expression. These results were confirmed by the kinetics of growth of the strains in liquid culture. To simulate the hypoglycemia conditions in the H. rolymorpha cells the medium with 0.1% glucose was used. It is found that the amount of NCYC 495 / SNCA cells in which the fluorescence-labeled alpha-synuclein was detected at the first 18 hours rapidly decreases at low glucose concentrations (0.1%) as compared to cells grown in media with a high concentrations of glucose (1%). The model yeast strains were more resistant to heterologous production of alpha-synuclein at the excess of glucose in the culture medium. It is established that alpha-synuclein toxicity for yeast model strains weakened by ions Mn2+; similarly toxic effect of Mn2+ ions in the environment is attenuated by the action of heterologous alpha-synuclein in the case when cells grown in media with low glucose (0.1%) concentration. The data show the effectiveness of the chosen model to study the effect of various exogenous factors and cytotoxicity of alpha-synuclein. Product Description popup.authors Бобак Ярослав Петрович Стасик Олег Володимирович Стасик Олена Георгіївна popup.nrat_date 2020-04-02 Close