Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Mar 25;11:1552. doi: 10.1038/s41467-020-15362-1

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 3 — a Schematic representation of the calyx of Held synapse with the cross-section revealing multiple swellings with active zones (AZ) and zoom-in delineating borders relevant for analysis. b Cartoon illustrating the topology of adjacent calyces as represented in c. c STORM images showing 16 targets of the same section acquired in 10 consecutive staining rounds (SR) using one or two spectral channels (Ch). d High registration precision (3–47 nm) of images shown in b achieved throughout all SRs. e Magnified images of cross-section of calyces (as depicted in a). Homer, bassoon, and WGA stainings confirm imaging plane-perpendicular orientation. f Presynaptic distribution of selected proteins analyzed and averaged over 9–15 calyces from 3 to 5 experiments. Calyx borders derived from WGA staining. g Distances of α-Tub peak, first and second F-Act peak (p1, p2), and VGlut1 peak to synaptic membrane differed significantly (**p ≤ 0.01; ***p ≤ 0.001, one-way ANOVA). h–j Differential protein distribution within regions containing or lacking AZs (see inset in h). h Synapse-proximal regions contain more synaptic vesicles (SVs) (VGlut1 staining) at AZ sites as compared with extra-AZ regions. i Less polymerized actin in AZ-proximal regions. j More MyoVa at AZ sites, correlating with VGlut1 profile. k, l Example of single AZs supports correlation between SV and MyoVa distribution and illustrates individuality of AZs. m Colocalization matrix showing Pearson’s r coefficients (exact values, see Supplementary Fig. 14) between all combinations of 14 analyzed proteins at AZ-proximal (upper matrix part) and AZ-free (lower part) calyx regions. Asterisks denote statistical significance for colocalization of selected protein pairs (*p ≤ 0.05; unpaired two-tailed Student’s t test). Scale bars correspond to 5 µm in c and 500 nm in e. Scatter plot g shows mean ± SD. Note that in addition to representative images in c and e another six and three experiments were carried out, respectively, with similar designs and outcomes. For exact sample numbers, see Supplementary Table 5. Source data, including exact p values for g and m, are provided as a Source Data file.