Abstract
GABAA receptors mediate synaptic and extrasynaptic inhibition. Native receptors consist of alpha and beta subunits, which are required for function, and another "modulatory" subunit, for example, gamma, delta, or epsilon. Of these, the epsilon subunit has the most restricted distribution, confers resistance to neurosteroid and anesthetic modulation, and causes spontaneous channel opening. Little is known, however, about how epsilon affects receptor kinetics, which in turn shape responses to both ambient and synaptic GABA exposure. Here, we expressed human alpha2beta1, alpha2beta1gamma2, or alpha2beta1epsilon subunit combinations in human embryonic kidney 293 cells and used rapid solution exchange to study receptor kinetics in outside-out patches. The epsilon subunit greatly slowed deactivation and recovery after brief GABA pulses. During long, saturating GABA pulses, the rate of desensitization was slower for alpha2beta1epsilon and alpha2beta1gamma2 than for alpha2beta1. However, in alpha2beta1epsilon, the final extent of desensitization was large compared with that of alpha2beta1gamma2. Responses in alpha2beta1epsilon, but not the others, were often followed by an "overshoot" above the baseline, suggesting that a fraction of channels are spontaneously open and are transiently silenced by receptor activation and subsequent desensitization. The baseline current and associated noise were reduced by picrotoxin, revealing that epsilon-containing channels are open approximately 4% of the time in the absence of GABA. These results suggest that, if epsilon-containing receptors are expressed at synapses, the synaptic currents would be long-lasting but may rundown quickly under high-frequency activation. In addition, silencing of spontaneous openings by desensitization raises the possibility that tonic inhibition mediated by epsilon-containing receptors may be regulated by phasic inhibition.