Fig. 6: Visualising the subsynaptic distribution of NTD-deleted receptors using 3D STORM imaging. | Nature Communications

Fig. 6: Visualising the subsynaptic distribution of NTD-deleted receptors using 3D STORM imaging.

From: AMPA receptor anchoring at CA1 synapses is determined by N-terminal ___domain and TARP γ8 interactions

Fig. 6

a Representative density plots of synaptic AMPAR clusters reconstructed as scatter plots, highlighting the nonuniform distribution of receptors (cold vs. hot colours corresponding to low vs. high densities). Observed organisation (top), versus random (bottom) re-distributions of measured localisations shown for comparison (Scale bars: 100 nm). b GluA1 ΔNTD receptors exhibited a small increase in nanocluster number per synapse compared to GluA1 full-length (numbers as median ± C.I., left, GluA1 full-length: 1 ± 1–1, GluA1 ΔNTD: 1.5 ± 1–2, p = 0.018, two-tailed Mann–Whitney test), whereas GluA2 nanocluster number was unaffected by NTD deletion (right, GluA2 full-length: 1 ± 1–1, GluA2 ΔNTD: 1 ± 1–1, p = 0.4895, two-tailed Mann–Whitney test). c Nanocluster diameters (longest axis) were increased for GluA1 ΔNTD (diameters (nm), left GluA1 full-length: 75.1 ± 68.0–80.6, GluA1 ΔNTD: 83.3 ± 74.1–90.8, p = 0.0459, two-tailed Mann–Whitney test) but unchanged for GluA2 ΔNTD receptors (diameters (nm), right GluA2 full-length: 83.7 ± 73.7-92.6, GluA2 ΔNTD: 78.1 ± 67.2-88.9, p = 0.2378, two-tailed Mann–Whitney test) relative to respective full-length controls. d GluA1 ΔNTD nanoclusters consisted of fewer localisations compared to GluA1 full-length receptors (left, GluA1 full-length: 42.0 ± 40.0–46.0, GluA1 ΔNTD: 38 ± 36.0–40.0, p = 0.0365, two-tailed Mann–Whitney test). The number of localisations per nanocluster was unchanged for GluA2 full-length vs ΔNTD (right, GluA2 full-length: 39.0 ± 37–41, GluA2 ΔNTD: 39.0 ± 36–44, p = 0.9189, two-tailed Mann–Whitney test). e NTD deletion of GluA1 (density (localisations/nanocluster), left, GluA1 full-length: 366166 ± 301447–436647, GluA1 ΔNTD: 235946 ± 209787-286802, p = 0.0005, two-tailed Mann–Whitney test) but not GluA2 (density (localisations/nanocluster), right GluA2 full-length: 211710 ± 179581–264751, GluA2 ΔNTD: 280631 ± 196833–370233, p = 0.1766, two-tailed Mann–Whitney test) reduced receptor densities within nanoclusters. Data shown as median ± lower-upper 95% CI, * indicates p < 0.05, ***p < 0.001 and ns specifies no significance. STORM data were obtained from 3 culture preparations for GluA1 (GluA1 full-length: 113 synapses/12 cells, GluA1 ΔNTD: 80 synapses/16 cells) and from four preparations for GluA2 (GluA2 full-length: 51 synapses/nine cells, GluA2 ΔNTD: 44 synapses/10 cells).

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