Pharmacological effects of the marine toxins, brevetoxin and saxitoxin, on murine frontal cortex neuronal networks

Abstract

Brevetoxins and saxitoxins (STXs), which are produced by marine dinoflagellates, are very potent neurotoxins targeting separate sites of the α subunit of voltage-dependent sodium channels (VDSCs). An attractive approach for marine toxin detection relies on pharmacological modulation of VDSCs expressed in cells or tissues. While these function-based cellular assays exhibit the required sensitivity, they are typically slow and have limited potential use for field applications. Cultured neuronal networks grown on substrate integrated microelectrode arrays (MEAs) have emerged as a robust and sensitive approach for environmental threat detection. The present work describes the rapid effects of brevetoxin-2 (PbTx-2) and STX on embryonic murine frontal cortex neuronal networks on MEAs. Network recording parameters such as mean spike rate, burst rate, burst duration, number of spikes per burst and spike amplitude were analyzed before and after exposure to the toxins. STX produced fast and reversible inhibition of all electrophysiological parameters with IC 50s ranging between 1.2 and 2.2 nM. Although PbTx-2 also caused inhibition of most of the network electrophysiological parameters, it produced an increase in burst duration at lower concentrations (EC 50=15±2 nM, n=4) followed by inhibition at higher ones (IC 50=63±4 nM, n=4). Exposure of frontal cortex networks to PbTx-2 and STX also caused differential effects on spike amplitude. This work demonstrates that cultured neuronal networks not only could be used for pharmacological characterization of marine toxins but they also provide a tool with unique properties for their detection.