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  • The Effect of DOPAL on the Viability, Reactivity and Bioenergetic Function of a Rat Astrocyte Cell Line

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  • DOPAL, a toxic, catecholaldehyde, dopamine metabolite, is hypothesized to induce neurodegeneration in Parkinson’s disease. However, the effect of DOPAL on astrocyte function is less well characterized. Astrocytes are often referred to as a ‘double-edged sword’ in Parkinson’s disease due to their conflicting protective and pathological effects on neurons.

    This project aimed to assess if exposure to DOPAL impaired the viability and bioenergetic function and induced reactivity of the Neu7 rat astrocyte cell line in vitro.

    alamarBlue and Picogreen assays, respectively, evaluated astrocyte metabolic activity and proliferation to determine the effect of DOPAL on Neu7 viability. To investigate if DOPAL induces astrocyte reactivity, hydrogen peroxide and nitrite production by Neu7 cells was determined. This was followed by immunohistochemical analysis of GFAP and AQP-4 expression by Neu7 cells (markers of astrocyte reactivity). Changes to astrocyte morphology in response to DOPAL treatment were evaluated using phalloidin staining. Bioenergetic Function was assessed using extracellular flux analysis with the Seahorse XFp analyser.

    Following an overnight treatment with DOPAL at concentrations of 500µM and 1000µM, astrocyte metabolic activity, proliferation and bioenergetic function significantly decreased compared to vehicle treated cells (p<0.05). Astrocytic hydrogen peroxide and nitrite production was significantly higher following overnight treatment with DOPAL at 500µM and 1000µM (p<0.05). 500µM of DOPAL altered astrocytic AQP-4 and GFAP expression levels and strongly decreased the fluorescence intensity of phalloidin- stained cells. Therefore, our results indicate that astrocyte viability was reduced and astrocyte reactivity was induced following DOPAL treatment.

    This is the first in vitro study showing that DOPAL is directly toxic to astrocytes. Astrocytes also acquire neurotoxic functions such as reactive oxygen species production and nitrite production in response to DOPAL treatment, which may contribute to neurodegeneration. This study provides a base for future research to elucidate the mechanism of DOPAL induced astrocyte toxicity in Parkinson’s disease.


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