Recruitment of Plasma Membrane GABA-A Receptors by Submembranous Gephyrin/Collybistin Clusters

It has been shown that subunit composition is the main determinant of the synaptic or extrasynaptic localization of GABA_A receptors (GABA_ARs). Synaptic and extrasynaptic GABA_ARs are involved in phasic and tonic inhibition, respectively. It has been proposed that synaptic GABA_ARs bind to the postsynaptic gephyrin/collybistin (Geph/CB) lattice, but not the typically extrasynaptic GABA_ARs. Nevertheless, there are no studies of the direct binding of various types of GABA_ARs with the submembranous Geph/CB lattice in the absence of other synaptic proteins, some of which are known to interact with GABA_ARs. We have reconstituted GABA_ARs of various subunit compositions, together with the Geph/CB scaffold, in HEK293 cells, and have investigated the recruitment of surface GABA_ARs by submembranous Geph/CB clusters. Results show that the typically synaptic α1β3γ2 GABA_ARs were trapped by submembranous Geph/CB clusters. The α5β3γ2 GABA_ARs, which are both synaptic and extrasynaptic, were also trapped by Geph/CB clusters. Extrasynaptic α4β3δ GABA_ARs consistently showed little or no trapping by the Geph/CB clusters. However, the extrasynaptic α6β3δ, α1β3, α6β3 (and less α4β3) GABA_ARs were highly trapped by the Geph/CB clusters. AMPA and NMDA glutamate receptors were not trapped. The results suggest: (I) in the absence of other synaptic molecules, the Geph/CB lattice has the capacity to trap not only synaptic but also several typically extrasynaptic GABA_ARs; (II) the Geph/CB lattice is important but does not play a decisive role in the synaptic localization of GABA_ARs; and (III) in neurons there must be mechanisms preventing the trapping of several typically extrasynaptic GABA_ARs by the postsynaptic Geph/CB lattice.