GABAA Receptor Binding and Ion Channel Function in Primary Neuronal Cultures for Neuropharmacology/Neurotoxicity Testing

Abstract

GABAA receptor (GABAAR) constitutes the main inhibitory receptor of the central nervous system. Due to the wide distribution and activity of its main neurotransmitter agonist, the γ-aminobutyric acid (GABA), its pharmacology has been thoroughly studied, given rise to the development of numerous drugs and of neuroactive compounds, some of the latest inducing neurotoxic effects. In this chapter, we describe methods for studying the interaction of chemical agents with the GABAAR and the effects they produce on its function, by using mice cortical and cerebellar granule neurons that have been grown in vitro (cultured neurons). The methods described here include the evaluation of the binding of different agents such as GABA agonists, GABA antagonists or allosteric modulators of the receptor, and the assessment of the receptor functionality by analyzing the Cl− flux induced by GABA.