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Human neuronal signaling and communication assays to assess functional neurotoxicity

Human neuronal signaling and communication assays to assess functional neurotoxicity

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LOSER, Dominik, Jasmin SCHAEFER, Timm DANKER, Clemens MÖLLER, Markus BRÜLL, Ilinca SUCIU, Anna-Katharina ÜCKERT, Stefanie KLIMA, Marcel LEIST, Udo KRAUSHAAR, 2021. Human neuronal signaling and communication assays to assess functional neurotoxicity. In: Archives of Toxicology. Springer. 95(1), pp. 229-252. ISSN 0340-5761. eISSN 1432-0738. Available under: doi: 10.1007/s00204-020-02956-3

@article{Loser2021-01Human-52078, title={Human neuronal signaling and communication assays to assess functional neurotoxicity}, year={2021}, doi={10.1007/s00204-020-02956-3}, number={1}, volume={95}, issn={0340-5761}, journal={Archives of Toxicology}, pages={229--252}, author={Loser, Dominik and Schaefer, Jasmin and Danker, Timm and Möller, Clemens and Brüll, Markus and Suciu, Ilinca and Ückert, Anna-Katharina and Klima, Stefanie and Leist, Marcel and Kraushaar, Udo} }

2020-12-10T13:33:13Z Danker, Timm Loser, Dominik Loser, Dominik Schaefer, Jasmin Prediction of drug toxicity on the human nervous system still relies mainly on animal experiments. Here, we developed an alternative system allowing assessment of complex signaling in both individual human neurons and on the network level. The LUHMES cultures used for our approach can be cultured in 384-well plates with high reproducibility. We established here high-throughput quantification of free intracellular Ca<sup>2+</sup> concentrations [Ca<sup>2+</sup>]i as broadly applicable surrogate of neuronal activity and verified the main processes by patch clamp recordings. Initially, we characterized the expression pattern of many neuronal signaling components and selected the purinergic receptors to demonstrate the applicability of the [Ca<sup>2+</sup>]i signals for quantitative characterization of agonist and antagonist responses on classical ionotropic neurotransmitter receptors. This included receptor sub-typing and the characterization of the anti-parasitic drug suramin as modulator of the cellular response to ATP. To exemplify potential studies on ion channels, we characterized voltage-gated sodium channels and their inhibition by tetrodotoxin, saxitoxin and lidocaine, as well as their opening by the plant alkaloid veratridine and the food-relevant marine biotoxin ciguatoxin. Even broader applicability of [Ca<sup>2+</sup>]i quantification as an end point was demonstrated by measurements of dopamine transporter activity based on the membrane potential-changing activity of this neurotransmitter carrier. The substrates dopamine or amphetamine triggered [Ca<sup>2+</sup>]i oscillations that were synchronized over the entire culture dish. We identified compounds that modified these oscillations by interfering with various ion channels. Thus, this new test system allows multiple types of neuronal signaling, within and between cells, to be assessed, quantified and characterized for their potential disturbance. Kraushaar, Udo Klima, Stefanie Möller, Clemens Klima, Stefanie Suciu, Ilinca Human neuronal signaling and communication assays to assess functional neurotoxicity eng Leist, Marcel Brüll, Markus Attribution 4.0 International Ückert, Anna-Katharina Möller, Clemens Kraushaar, Udo Brüll, Markus Leist, Marcel 2021-01 Schaefer, Jasmin 2020-12-10T13:33:13Z Ückert, Anna-Katharina Suciu, Ilinca Danker, Timm

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