Modulation of spontaneous and GABA-evoked tonic alpha4beta3delta and alpha4beta3gamma2L GABAA receptor currents by protein kinase A.

Protein kinase A (PKA) has been reported to regulate synaptic alphabetagamma gamma-aminobutyric acid type A (GABA(A)) receptor currents, but whether PKA regulates GABA(A) receptor peri- and extrasynaptic tonic currents is unknown. GABA(A) receptors containing alpha4 subunits are important in mediating tonic inhibition and exist as both alpha4betadelta and alpha4betagamma receptors in the brain. To mimic GABA-independent and GABA-dependent tonic currents, we transfected HEK 293T cells with alpha4beta3delta or alpha4beta3gamma2L subunits and recorded spontaneous currents in the absence of applied GABA and steady-state currents in the presence of 1 muM GABA. Both alpha4beta3delta and alpha4beta3gamma2L receptors displayed spontaneous currents, but PKA activation increased spontaneous alpha4beta3delta currents substantially more than spontaneous alpha4beta3gamma2L currents. The increase in spontaneous alpha4beta3delta currents was due to an increase in single-channel open frequency. In contrast, PKA activation did not alter steady-state tonic currents recorded in the presence of 1 muM GABA. We concluded that PKA had a GABA concentration-dependent effect on alpha4beta3delta and alpha4beta3gamma2L currents. In the absence of GABA, spontaneous alpha4beta3delta and, to a lesser extent, alpha4beta3gamma2L currents could provide a basal, tonic current that could be regulated by PKA. With increasing concentrations of extracellular GABA, however, tonic alpha4beta3delta and alpha4beta3gamma2L currents would become more GABA dependent and less PKA sensitive. Thus in brain regions with fluctuating extracellular GABA levels, the dynamic range of GABA-activated tonic currents would be set by PKA and the increase in tonic current produced by increasing GABA would be reduced by PKA-mediated phosphorylation. When ambient GABA reaches micromolar concentrations, PKA would have no effect on steady-state tonic currents.