Stargazin modulates native AMPA receptor functional properties by two distinct mechanisms

AMPA receptors play a central role in basal excitatory synaptic transmission as well as synaptic maturation and plasticity. The transmembrane AMPA receptor regulatory protein (TARP) stargazin (gamma 2) serves multiple roles in trafficking and stabilizing synaptic AMPA receptors and may be incorporated as an auxiliary subunit. We wanted to determine whether stargazin altered channel function of neuronal AMPA receptors. Transfection of cultured hippocampal neurons with stargazin produced two distinct effects on AMPA receptor functional properties: a sixfold reduction in glutamate-evoked desensitization and a twofold increase in the relative size of responses to the partial agonist kainate. Kinetic and dose-response analyses suggest that the effect of stargazin on glutamate desensitization results from an allosteric interaction that destabilizes the desensitized state of the receptor and that potentiation of kainate responses reflects increased efficacy rather than a change in affinity. These functional effects were also observed in human embryonic kidney 293 cells transfected with various heteromeric and homomeric AMPA receptors, with distinct subunit-dependent effects on glutamate desensitization, kainate efficacy, and trafficking. Two regions of stargazin mediate its functional effects: the C-terminal intracellular domain seems to be more important for effects on glutamate-evoked desensitization and receptor trafficking, whereas the first extracellular domain makes a larger contribution to effects on kainate efficacy. These data indicate that TARPs are involved both in trafficking and direct modulation of channel function and, as auxiliary subunits of neuronal AMPA receptors, must be considered in the functional heterogeneity of neuronal AMPA receptors.