Silencing of spontaneous activity at α4β1/3δ GABA_A receptors in hippocampal granule cells reveals different ligand pharmacology.

ABSTRACT:

Background and purpose

The δ-subunit-containing GABA_A receptors, α₄ β₁ δ and α₄ β₃ δ, in dentate gyrus granule cells (DGGCs) are known to exhibit both spontaneous channel openings (i.e. constitutive activity) and agonist-induced current. The functional implications of spontaneous gating are unclear. In this study, we tested the hypothesis that constitutively active α₄ β_1/3 δ receptors limit agonist efficacy.

Experimental approach

Whole-cell electrophysiological recordings of adult male rat and mouse hippocampal DGGCs were used to characterize known agonists and antagonists at δ-subunit-containing GABA_A receptors. To separate constitutive and agonist-induced currents, different recording conditions were employed.

Key results

Recordings at either 24°C or 34°C, including the PKC autoinhibitory peptide (19-36) intracellularly, removed spontaneous gating by GABA_A receptors. In the absence of spontaneous gating, DGGCs responded to the α₄ β_1/3 δ orthosteric agonist Thio-THIP with a four-fold increased efficacy relative to recording conditions favouring constitutive activity. Surprisingly, the neutral antagonist gabazine was unable to antagonize the current by Thio-THIP. Furthermore, a current was elicited by gabazine alone only when the constitutive current was silenced (EC₅₀ 2.1 μM). The gabazine-induced current was inhibited by picrotoxin, potentiated by DS2, completely absent in δ^-/- mice and reduced in β₁ ^-/- mice, but could not be replicated in human α₄ β_1/3 δ receptors expressed heterologously in HEK cells.

Conclusion and implications

Kinase activity infers spontaneous gating in α₄ β_1/3 δ receptors in DGGCs. This significantly limits the efficacy of GABA_A agonists and has implications in pathologies involving aberrant excitability caused by phosphorylation (e.g. addiction and epilepsy). In such cases, the efficacy of δ-preferring GABA_A ligands may be reduced.

SUBMITTER: Dalby NO 

PROVIDER: S-EPMC7429488 | biostudies-literature |

REPOSITORIES: biostudies-literature