Nanoscale distribution of β-secretase in the functional zones of excitatory postsynapse using STED microscopy
(A and B) Overlay of STED images of postsynaptic density marker PSD95 and a marker for endocytic zone Dynamin (green) with β-secretase (Magenta). The black contour indicates the automated detection of neuronal processes. Inset 1–6 indicate a gallery of synapses where the black contour within inset represents automatically detected regions for confocal marker for postsynapse and endocytic zone (black). Black in the overlay images represents the overlap between the corresponding green and magenta images. The scale bars in B represent 7 μm and inset corresponds to 1.4 μm.
(i and iv) Comparison of RSP and RSE for quantifying colocalization of β-secretase for functional zones of an excitatory postsynapse. Data are represented as mean ± SEM. Significance was determined by unpaired two-tailed Student's t test. ∗p ≤ 0.05, ∗∗p ≤ 0.01, and ∗∗∗p ≤ 0.001, ns p > 0.05.
(ii and iii) Indicate the distribution of length of all β-secretase nanodomains obtained by STED microscopy in post and perisynaptic compartments, respectively.
(v and vi) Indicate the distribution of intensity of all β-secretase nanodomains obtained by STED microscopy in post and perisynaptic compartments, respectively. n = 5669 puncta (post) and 3798 puncta (peri).