title: Multiparametric characterisation of neuronal network activity for in vitro agrochemical neurotoxicity assessment.
abstract: The last few decades have seen the marketing of hundreds of new pesticide products with a forecasted expansion of the global agrochemical industry. As several pesticides directly target nervous tissue as their mechanism of toxicity, alternative methods to routine in vivo animal testing, such as the Multi Electrode Array (MEAs)-based approach, have been proposed as an in vitro tool to perform sensitive, quick and low cost neuro-toxicological screening. Here, we examined the effects of a training set of eleven active substances known to have neuronal or non-neuronal targets, contained in the most commonly used agrochemicals, on the spontaneous electrical activity of cortical neuronal networks grown on MEAs. A multiparametric characterisation of neuronal network firing and bursting was performed with the aim of investigating how this can contribute to the efficient evaluation of in vitro chemical-induced neurotoxicity. The analysis of MFR, MBR, MBD, MISI_B and % Spikes_B parameters identified four different groups of chemicals: one wherein only inhibition is observed (chlorpyrifos, deltamethrin, orysastrobin, dimoxystrobin); a second one in which all parameters, except the MISI_B, are inhibited (carbaryl, quinmerac); a third in which increases at low chemical concentration are followed by decreases at high concentration, with exception of MISI_B that only decreased (fipronil); a fourth in which no effects are observed (paraquat, glyphosate, imidacloprid, mepiquat). The overall results demonstrated that the multiparametric description of the neuronal networks activity makes MEA-based screening platform an accurate and consistent tool for the evaluation of the toxic potential of chemicals. In particular, among the bursting parameters the MISI_B was the best that correlates with potency and may help to better define chemical toxicity when MFR is affected only at relatively high concentration.
authors.affiliation: ETT Spa, via Sestri 37, 16154 Genoa, Italy; Institute of Biophysics, National Research Council, via De Marini 6, 16149 Genoa, Italy. Electronic address: susanna.alloisio@ettsolutions.com.
authors.forename: Susanna
authors.initials: S
authors.lastname: Alloisio
authors.affiliation: Institute of Biophysics, National Research Council, via De Marini 6, 16149 Genoa, Italy. Electronic address: nobile@ge.ibf.cnr.it.
authors.forename: Mario
authors.initials: M
authors.lastname: Nobile
authors.affiliation: ETT Spa, via Sestri 37, 16154 Genoa, Italy; Institute of Biophysics, National Research Council, via De Marini 6, 16149 Genoa, Italy. Electronic address: antonio.novellino@ettsolutions.com.
authors.forename: Antonio
authors.initials: A
authors.lastname: Novellino
mesh_terms: D000200:Action Potentials; D016573:Agrochemicals / Q000145:classification / Q000633:toxicity*; D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects* / Q000196:embryology / Q000503:physiopathology; D004305:Dose-Response Relationship, Drug; D005865:Gestational Age; D057166:High-Throughput Screening Assays; D006801:Humans; D009415:Nerve Net / Q000187:drug effects* / Q000503:physiopathology; D020258:Neurotoxicity Syndromes / Q000209:etiology* / Q000503:physiopathology; D017208:Rats, Wistar; D018570:Risk Assessment; D012815:Signal Processing, Computer-Assisted; D013997:Time Factors; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D016573:Agrochemicals
keywords: In vitro neurotoxicity; Micro electrode array; Neuronal bursting behaviour; Pesticides; Spontaneous electrical activity