Synaptic density quantifications were performed as described previously [17] using our in-house Synapse Counter plugin for ImageJ (freely available at https://github.com/SynPuCo/SynapseCounter). Briefly, the structurally complete glutamatergic synapses were identified by the spatial overlap of presynaptic marker VGLUT1 and postsynaptic markers PSD95. For each condition, 56 micrographs were obtained (n = 4) using the Carl Zeiss LSM 710 confocal microscope using the alpha Plan-Apochromat × 100 oil objective. The densities of synaptic puncta were quantified in 66.5 × 66.5 μm regions containing a single neuron perikaryon.
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