Alteration of GABAergic synapses and gephyrin clusters in the thalamic reticular nucleus of GABA_A receptor α3 subunit-null mice

Multiple GABA_A-receptor subtypes are assembled from α, β and γ subunit variants. GABA_A receptors containing the α3 subunit represent a minor population with a restricted distribution in the CNS. In addition, they predominate in monoaminergic neurons and in the nucleus reticularis thalami (nRT), suggesting a role in the regulation of cortical function and sleep. Mice with a targeted deletion of the α3 subunit gene (α3^0/0) are viable and exhibit a subtle behavioural phenotype possibly related to dopaminergic hyperfunction. Here, we investigated immunohistochemically the consequences of the loss of α3 subunit for maturation of GABA_A receptors and formation of GABAergic synapses in the nRT. Throughout postnatal development, the regional distribution of the α1, α2, or α5 subunit was unaltered in α3 ^0/0 mice and the prominent α3 subunit staining of nRT neurons in wildtype mice was not replaced. Subcellularly, as seen by double immunofluorescence, the α3 and γ2 subunit were clustered at postsynaptic sites in the nRT of adult wildtype mice along with the scaffolding protein gephyrin. In α3^0/0 mice, γ2 subunit clustering was disrupted and gephyrin formed large aggregates localized at the cell surface, but unrelated to postsynaptic sites, indicating that nRT neurons lack postsynaptic GABA_A receptors in mutant mice. Furthermore, GABAergic terminals were enlarged and reduced in number, suggesting a partial deficit of GABAergic synapses. Therefore, GABA_A receptors are required for gephyrin clustering and long-term synapse maintenance. The absence of GABA _A-mediated transmission in the nRT may have a significant impact on the function of the thalamo-cortical loop of α3^0/0 mice.