Colocalization of gephyrin and GABAA‐receptor subunits in the rat retina

Abstract

Gephyrin is a protein that copurifies with the glycine receptor (GlyR) and is required for the clustering of GlyRs at postsynaptic sites. Previously, it was thought that antibody mAb 7a, directed against gephyrin, was a specific marker for GlyR. However, there is evidence that gephyrin can also be found at nonglycinergic synapses. Here, immunocytochemistry was applied to show this directly for the rat retina. Both gephyrin and different Subunits of the γ‐aminobutyric acid (GABA)_A receptor were localized to discrete puncta in the inner plexiform layer, and these puncta were shown by electron microscopy to represent synaptic sites. Double immunocytochemistry revealed that GABA_A receptors and GlyRs are not colocalized. However, gephyrin and different subunits of GABA_A receptors were found to occur at the same synapses. The amount of colocalization varied with the GABA_A receptor subunit composition and was most extensive for the α2 subunit, less for the α3 subunit, and minimal for the αl subunit. The gephyrin present at GABAergic synapses of the retina might also be involved with clustering of receptors at the postsynaptic sites. Hence, localization of gephyrin can no longer be considered as a unique marker of glycinergic synapses.