N-cadherin has a spectrum of localization patterns at the synaptic cleft.

<p>(<b>A</b>) N-cadherin is localized between the pre- and post-synaptic compartments, represented by vGlut1 and PSD95 respectively. (<b>B</b>) N-cadherin is localized at or adjacent to the active zone, represented by bassoon. <i>Arrows</i>, N-cadherin puncta associated with synapses; <i>arrowheads</i>, N-cadherin puncta not associated with synapses. (<b>C</b>) N-cadherin localization at synapses varies from punctate, often flanking one side of the synapse, to uniform along the synaptic cleft. (<b>D</b>) Classification of N-cadherin localization patterns relative to bassoon into five categories (single puncta, double puncta, cleft with puncta, cleft, and round(bassoon)/puncta). Representative images and schematics of the five different N-cadherin localization patterns. (<b>E</b>) Percentage of synapses (mean±s.e.m) in 17-20 DIV hippocampal neurons in each N-cadherin pattern category. n=7 experiments, ≥77 synapses per experiment, 650 synapses total. p<0.0001, one-way ANOVA, Tukey’s post-test (p<0.01 for single puncta vs. double puncta; p<0.01 for single puncta vs. cleft; p>0.05 for single puncta vs. cleft with puncta; p<0.001 for single puncta vs. round/puncta). (<b>F</b>) Percentage of synapses (mean±s.e.m) in each N-cadherin pattern category in 11 DIV hippocampal neurons compared to matched cultures at 17-20 DIV. N-cadherin at synapses from neurons at 11 DIV is distributed more evenly along the synaptic cleft and less as puncta compared to synapses from neurons at 17-20 DIV. Two-way ANOVA with matched values and Bonferroni post-test. n=2 experiments, ≥103 synapses per experiment.