Counting NMDA Receptors at the Cell Surface

In the mammalian central nervous system, N-methyl-d-aspartate (NMDA) receptors mediate fast excitatory synaptic transmission. These receptors also play a key role in brain development, synaptic plasticity, and memory formation, and their dysregulation can cause neurological and/or psychiatric disorders. A variety of techniques have been applied to examine the molecular mechanisms that regulate the number of NMDA receptors at the cell surface. Here we provide a step-by-step guides for two commonly used techniques: a quantitative assay that measures both surface and total receptor pools, and a biotinylation assay that can rapidly evaluate the surface and total expression levels of receptor subunits. Among other applications, these methods can be used to measure the cell surface expression of recombinant receptors with genetic alterations and the effect of co-expressing additional proteins that may regulate subunit surface trafficking. These techniques can be used either individually or in parallel experiments, thus allowing researchers to independently validate observations regarding the regulation of NMDA receptor expression.
Source: Springer protocols feed by Neuroscience - Category: Neuroscience Source Type: news