12 well microelectrode array mea plates

To measure neuronal firing activity, ARCOs at 60 DIV were plated onto 12-well microelectrode array (MEA) plates (Axion Biosystems) precoated with Matrigel and maintained in BrainPhys neuronal culture media (Stemcell Technologies). Data of spontaneous neuronal firing were collected for 120 s in 37°C at 2 weeks after plating using the default neural activity settings from the manufacturer (Axion Biosystems Maestro Axis software version 2.4.2.13) (Wang et al., 2019 ). Mean firing rate was measured by counting spikes from active electrodes in each well per minute. Spike and burst detection were further analyzed and plotted by Neural Metric Tool version 2.2.3.

Neuronal network field electrical spike activity (ESA), an aggregate measure of extracellular field potentials spontaneously produced by the neuronal networks, was recorded using NGCs cultured on 12-well microelectrode array (MEA) plates (Axion BioSystems, Atlanta, GA), as detailed in the Supporting Information and previously described.^{35 (link)} Briefly, the ESA was measured from 7 DIV NGCs with the acute addition of the vehicle or the indicated concentrations of TBP using the Axis software (Version 2.4, Axion BioSystems), with specific settings and data analysis software described in the Supporting Information.