Interaction of insecticides with heteromeric GABA-gated chloride channels from zebrafish Danio rerio (Hamilton)

The ionotropic GABA_A receptor (GABA_AR) is the main fast inhibitory post-synaptic receptor and is also an important insecticidal target. Effect of insecticides on fish has attracted intensive attention. However, no systematic study on heteromeric zebrafish GABA_AR expressed in oocytes has been reported to date. In this study, the α1 subunit, the β2S subunit lacking 47 amino acid residues compared with the β2L subunit, and the γ2 subunit having five transmembrane domains were isolated from zebrafish Danio rerio. The responses of homomeric and heteromeric (α1, β2S and γ2) channels to agonists and GABA_AR-targeted compounds were detected with two-electrode voltage clamp. Dose-dependent responses were observed in heteromeric α1β2S, β2Sγ2, and α1β2Sγ2 GABAR channels with EC₅₀ values at 21.75, 6291, and 33.69 μM for GABA-induced current and 3.28, 155.5, and 3.79 mM for β-alanine-induced current, respectively. However, no response was induced by benzamidine in all GABAR channels. Abamectin, dieldrin, fluralaner and fipronil could strongly inhibited GABA-induced inward current ≥50% at 10^-6 M, while α-endosulfan, flufiprole and ethiprole only inhibited GABA-induced current <50%. This study has clarified the interaction of insecticides with the heteromeric GABA_AR channel, which could help us further explore the potential function and toxicological importance of GABA_ARs from D. rerio.