Gephyrin selective intrabodies as a new strategy for studying inhibitory receptor clustering

摘要: The microtubule-binding protein gephyrin is known to play a pivotal role in targeting and clustering postsynaptic inhibitory receptors. Here, the Intracellular Antibodies Capture Technology (IATC) was used select two single-chain antibody fragments or intrabodies, which, fused nuclear localization signals (NLS), were able efficiently selectively remove from glycine receptor (GlyR) clusters. Co-transfection of NLS-tagged individual intrabodies with gephyrin-enhanced green fluorescent (EGFP) HEK 293 cells revealed partial relocalization aggregates onto nucleus perinuclear area. When expressed cultured neurons, these caused significant reduction number immunoreactive GlyR clusters, which associated decrease peak amplitude glycine-evoked whole cell currents as assessed electrophysiological experiments. Hampering function at posttranslational level may represent an attractive alternative for interfering more spatially localized manner.