Quercetin antagonism of GABAAρ1 receptors is prevented by ascorbic acid through a redox-independent mechanism

• Published in: European journal of pharmacology Vol. 714; no. 1-3; pp. 274 - 280
• Main Authors: Calero, Cecilia I., Beltrán González, Andrea N., Gasulla, Javier, Alvarez, Silvia, Evelson, Pablo, Calvo, Daniel J.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15-08-2013
• Subjects: Allosteric modulator; Allosteric Regulation - drug effects; Animals; Ascorbic acid; Ascorbic Acid - pharmacology; binding sites; Dose-Response Relationship, Drug; Flavonoid; frogs; GABA receptor; GABA-A Receptor Antagonists - pharmacology; gamma-aminobutyric acid; Histidine - metabolism; Humans; ion channels; mutants; oocytes; pharmacology; Quercetin; Quercetin - antagonists & inhibitors; Quercetin - pharmacology; receptors; Receptors, GABA-A - chemistry; Receptors, GABA-A - metabolism; site-directed mutagenesis; thiols; Ascorbic acid; NEM; Quercetin; GABA; GABA receptor; Allosteric modulator; Flavonoid; N-ethylmaleimide; γ-aminobutyric acid
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2013.07.044

Quercetin is a natural flavonoid widely distributed in plants that acts as a neuroprotective agent and modulates the activity of different synaptic receptors and ion channels, including the ionotropic GABA receptors. GABAAρ1 receptors were shown to be antagonized by quercetin, but the mechanisms underlying these antagonistic actions are still unknown. We have analyzed here if the antagonistic action produced by quercetin on GABAAρ1 receptors was related to its redox activity or due to alternative mechanism/s.
Homomeric GABAAρ1 receptors were expressed in frog oocytes and GABA-evoked responses electrophysiologically recorded. Quercetin effects on GABAAρ1 receptors were examined in the absence or presence of ascorbic acid. Chemical protection of cysteines by selective sulfhydryl reagents and site directed mutagenesis experiments were also used to determine ρ1 subunit residues involved in quercetin actions.
Quercetin antagonized GABAAρ1 receptor responses in a dose-dependent, fast and reversible manner. Quercetin inhibition was prevented in the presence of ascorbic acid, but not by thiol reagents that modify the extracellular Cys-loop of these receptors. H141, an aminoacidic residue located near to the ρ1 subunit GABA binding site, was involved in the allosteric modulation of GABAAρ1 receptors by several agents including ascorbic acid. Quercetin similarly antagonized GABA-evoked responses mediated by mutant H141DGABAAρ1 and wild-type receptors, but prevention exerted by ascorbic acid on quercetin effects was impaired in mutant receptors. Taken together the present results suggest that quercetin antagonistic actions on GABAAρ1 receptors are mediated through a redox-independent allosteric mechanism.