Allosteric Modulation of Retinal GABA Receptors by Ascorbic Acid

Abstract

Ionotropic GABA receptors (GABA_A and GABA_C) belong to the Cys-loop receptor family of ligand-gated ion channels. GABA_C receptors are highly expressed in the retina, mainly localized at the axon terminals of bipolar cells. Ascorbic acid, an endogenous redox agent, modulates the function of diverse proteins, and basal levels of ascorbic acid in the retina are very high. However, the effect of ascorbic acid on retinal GABA receptors has not been studied. Here we show that the function of GABA_C and GABA_A receptors is regulated by ascorbic acid. Patch-clamp recordings from bipolar cell terminals in goldfish retinal slices revealed that GABA_C receptor-mediated currents activated by tonic background levels of extracellular GABA, and GABA_C currents elicited by local GABA puffs, are both significantly enhanced by ascorbic acid. In addition, a significant rundown of GABA puff-evoked currents was observed in the absence of ascorbic acid. GABA-evoked Cl⁻ currents mediated by homomeric ρ₁ GABA_C receptors expressed in Xenopus laevis oocytes were also potentiated by ascorbic acid in a concentration-dependent, stereo-specific, reversible, and voltage-independent manner. Studies involving the chemical modification of sulfhydryl groups showed that the two Cys-loop cysteines and histidine 141, all located in the ρ₁ subunit extracellular domain, each play a key role in the modulation of GABA_C receptors by ascorbic acid. Additionally, we show that retinal GABA_A IPSCs and heterologously expressed GABA_A receptor currents are similarly augmented by ascorbic acid. Our results suggest that ascorbic acid may act as an endogenous agent capable of potentiating GABAergic neurotransmission in the CNS.