Topic: surface expression

AIMS:Investigate whether the endoplasmic reticulum (ER) quality control machinery verifies the structure of ligand-binding domains (LBDs) of NMDARs or controls the functionality of NMDARs.METHODS:HEK293 cells were used to express wild-type (WT) or mutation forms of GluN1/GluN2A and GluN1/GluN2B receptors. Surface expressions were observed using fluorescence confocal microscopy. Electrophysiological properties of NMDARs were assessed via the whole-cell patch-clamp technique.RESULTS:Using alanine substitutions combined with microscopy and electrophysiology, we found that surface expression of Gl

Maintaining the delicate balance between excitatory and inhibitory synaptic activity is crucial for proper brain function. The endosomal trafficking complex, retromer, plays a pivotal role in synaptic function by regulating the post-endocytic sorting of a variety of integral proteins at both excitatory and inhibitory synapses. As a result, SNX27/retromer is emerging as a potential mediator of excitatory/inhibitory crosstalk. In surface proteomic screens from primary cultured neurons, we have shown that a variety of subunits of γ-aminobutyric acid receptors (GABAARs) exhibit reduced surface exp

AIMS: Investigate the impact of disrupting specific disulfide bonds in the GluN1 subunit on the early trafficking and functional properties of conventional NMDARs.METHODS:HEK293 cells were utilized to express wild-type (WT) or mutant forms of GluN1/GluN2A and GluN1/GluN2B receptors. Surface expression and colocalization of NMDAR with the Golgi apparatus were observed using fluorescence confocal microscopy. Electrophysiological properties of NMDARs were assessed via the whole-cell patch-clamp technique. In addition, NMDA-induced excitotoxicity in rat hippocampal neurons was examined specificall

AIMS: Investigate the impact of disrupting specific disulfide bonds in the GluN1 subunit on the early trafficking and functional properties of unconventional NMDARs.METHODS:HEK293 cells and rat hippocampal neurons were used to express wild-type (WT) or mutant forms of GluN1/GluN3A receptors. Surface expression and colocalization of NMDARs with the Golgi apparatus (GA) were observed using fluorescence confocal microscopy. Electrophysiological properties of NMDARs were assessed via patch-clamp technique.RESULTS:Disrupting disulfide bonds in the GluN1 subunit co-transfected with GluN3A subunit le

N-methyl-D-aspartate receptors (NMDARs) are a family of ionotropic glutamate receptors that mediate excitatory neurotransmission in the CNS. Native NMDARs are assembled as heterotetramers composed of two obligatory GluN1 and two GluN2A-D/GluN3A-B subunits at various combinations. The opening of the ion channel is a central feature of NMDAR function and is controlled by binding of two co-agonists (glutamate/glycine) to the agonist binding domains (ABD). In our previous work, using molecular dynamics simulations, we proposed functionally important residues in this process of channel opening, Glu