title: Editor's Highlight: Evaluation of a Microelectrode Array-Based Assay for Neural Network Ontogeny Using Training Set Chemicals.
abstract: Thousands of compounds in the environment have not been characterized for developmental neurotoxicity (DNT) hazard. To address this issue, methods to screen compounds rapidly for DNT hazard evaluation are necessary and are being developed for key neurodevelopmental processes. In order to develop an assay for network formation, this study evaluated effects of a training set of chemicals on network ontogeny by measuring spontaneous electrical activity in neural networks grown on microelectrode arrays (MEAs). Rat (0-24 h old) primary cortical cells were plated in 48 well-MEA plates and exposed to 6 compounds: acetaminophen, bisindolylmaleimide-1 (Bis-1), domoic acid, mevastatin, sodium orthovanadate, and loperamide for a period of 12 days. Spontaneous network activity was recorded on days 2, 5, 7, 9, and 12 and viability was assessed using the Cell Titer Blue assay on day 12. Network activity (e.g. mean firing rate [MFR], burst rate [BR], etc), increased between days 5 and 12. Random Forest analysis indicated that across all compounds and times, temporal correlation of firing patterns (r), MFR, BR, number of active electrodes and % of spikes in a burst were the most influential parameters in separating control from treated wells. All compounds except acetaminophen (≤ 30 µM) caused concentration-related effects on one or more of these parameters. Domoic acid and sodium orthovanadate altered several of these parameters in the absence of cytotoxicity. Although cytotoxicity was observed with Bis1, mevastatin, and loperamide, some parameters were affected by these compounds at concentrations below those resulting in cytotoxicity. These results demonstrate that this assay may be suitable for screening of compounds for DNT hazard identification.
authors: NHEERL, US EPA, Research Triangle Park, NC, USA.
authors: Jasmine P
authors: JP
authors: Brown
authors: NHEERL, US EPA, Research Triangle Park, NC, USA.
authors: Diana
authors: D
authors: Hall
authors: NHEERL, US EPA, Research Triangle Park, NC, USA.
authors: Christopher L
authors: CL
authors: Frank
authors: NHEERL, US EPA, Research Triangle Park, NC, USA.
authors: Kathleen
authors: K
authors: Wallace
authors: NHEERL, US EPA, Research Triangle Park, NC, USA.
authors: William R
authors: WR
authors: Mundy
authors: NHEERL, US EPA, Research Triangle Park, NC, USA shafer.tim@epa.gov.
authors: Timothy J
authors: TJ
authors: Shafer
mesh_terms: D000082:Acetaminophen; D000818:Animals; D007211:Indoles; D007608:Kainic Acid / Q000031:analogs & derivatives; D008139:Loperamide; D008148:Lovastatin / Q000031:analogs & derivatives; D008301:Maleimides; D008839:Microelectrodes; D009415:Nerve Net / Q000187:drug effects*; D009474:Neurons / Q000187:drug effects*; D061251:Primary Cell Culture; D051381:Rats; D018675:Toxicity Tests / Q000379:methods*; D014638:Vanadates
chemical_list: D007211:Indoles; D008301:Maleimides; C012258:mevastatin; D000082:Acetaminophen; D014638:Vanadates; D008139:Loperamide; D008148:Lovastatin; C012301:domoic acid; C088060:bisindolylmaleimide; D007608:Kainic Acid
keywords: alternatives to animal testing; developmental; electrophysiology; in vitro and alternatives; neurotoxicity; neurotoxicology; screening.