Assessment of in vitro dopamine-neuroblastoma cell interactions with a bioelectric biosensor: perspective for a novel in vitro functional assay for dopamine agonist/antagonist activity

Publication date: 1 August 2017
Source:Talanta, Volume 170
Author(s): Theofylaktos Apostolou, Georgia Moschopoulou, Evdokia Kolotourou, Spyridon Kintzios
Current receptor-binding assays for dopamine do not measure the in vitro whole cellular response against dopamine or potential agonist/antagonist molecules. We herewith report the development of a novel functional assay concept for studying the in vitro interaction of the neurotransmitter dopamine with neural cells bearing dopamine receptors. The concept is based on the ultra-rapid measurement of changes in the electric properties of cultured N2a mouse neuroblastoma cells (corresponding to cumulative changes of the cell membrane potential). A close relationship between cumulative cell membrane potential and dopamine concentration was observed. Membrane depolarization was observed at nanomolar dopamine concentrations, while hyperpolarization was associated with micromolar ones. Treatment with the dopamine D2-receptor antagonist eticlopride resulted to a concentration-dependent membrane depolarization. Treatment with sodium chloride caused considerable weakening of the dopamine-associated hyperpolarization effect. The observed bioelectric response to dopamine was highly inversely correlated with the pattern of dopamine release-uptake balance by N2a cells, as determined with cyclic voltammetry. The bioelectric approach was also used to evaluate the dopaminergic activity of chaste tree (Vitex agnus-castus) extracts. The novel assay concept offers promising perspectives for the development of advanced companion diagnostics system for the high throughput, fast functional characterization of neurotransmitter agonists and antagonists.

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