Presynaptic alpha(2)delta subunits are key organizers of glutamatergic synapses

Schoepf CL, Ablinger C, Geisler SM, Stanika R, Campiglio M, Kaufmann WA, Nimmervoll B, Schlick B, Brockhaus J, Missler M, Shigemoto R, Obermair GJ

Abstract

In nerve cells the genes encoding for alpha(2)delta subunits of voltage-gated calcium channels have been linked to synaptic functions and neurological disease. Here we show that alpha(2)delta subunits are essential for the formation and organization of glutamatergic synapses. Using a cellular alpha(2)delta subunit triple-knockout/knockdown model, we demonstrate a failure in presynaptic differentiation evidenced by defective presynaptic calcium channel clustering and calcium influx, smaller presynaptic active zones, and a strongly reduced accumulation of presynaptic vesicle-associated proteins (synapsin and vGLUT). The presynaptic defect is associated with the downscaling of postsynaptic AMPA receptors and the postsynaptic density. The role of alpha(2)delta isoforms as synaptic organizers is highly redundant, as each individual alpha(2)delta isoform can rescue presynaptic calcium channel trafficking and expression of synaptic proteins. Moreover, alpha(2)delta-2 and alpha(2)delta-3 with mutated metal ion-dependent adhesion sites can fully rescue presynaptic synapsin expression but only partially calcium channel trafficking, suggesting that the regulatory role of alpha(2)delta subunits is independent from its role as a calcium channel subunit. Our findings influence the current view on excitatory synapse formation. First, our study suggests that postsynaptic differentiation is secondary to presynaptic differentiation. Second, the dependence of presynaptic differentiation on alpha(2)delta implicates alpha(2)delta subunits as potential nucleation points for the organization of synapses. Finally, our results suggest that alpha(2)delta subunits act as trans-synaptic organizers of glutamatergic synapses, thereby aligning the synaptic active zone with the postsynaptic density.