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. 2022 Apr 28;13:2297. doi: 10.1038/s41467-022-29751-1

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© The Author(s) 2022, corrected publication 2023

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/.

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Fig. 2 — a Representative images of Tenm3 nanoclusters (red) and presynaptic Bassoon macroclusters (green) show that the majority of Tenm3 nanoclusters in the proximal CA1 region are localized close to presynaptic active zones. Left images show overviews; right images, zoom-ins, in the whole field (left) and zoon-in field (right) at proximal CA1. Mice were infected with AAVs at P0 and analyzed at P13 (arrows = synaptic Tenm3 nanoclusters co-localized with Bassoon macroclusters; asterisks = non-synaptic Tenm3 nanoclusters that are >80 nm separated from presynaptic Bassoon macroclusters). b The Tenm3 deletion suppresses synaptic Tenm3 nanoclusters in the proximal CA1 region. Summary graphs show the percentage of Tenm3 nanoclusters that are synaptic (top) or non-synaptic (bottom) in control-infected (black) or Cre-expressing AAVs Tenm3 cKO mice (red). c Representative images of individual synapses in the proximal CA1 containing Tenm3 nanoclusters in mice without (ΔCre, left) and with deletion of Tenm3 in the MEC (Cre, right). d–f Same as a–c, except that brain sections were analyzed from distal subiculum. g Tenm3 nanoclusters occupy approximately 6% of the presynaptic active zone. Summary graphs depict the radius of Tenm3 nanoclusters (left) and Bassoon macroclusters (right) in the proximal CA1 and distal subiculum. h Most (>90%) presynaptic active zones visualized as Bassoon macroclusters contain 1–2 Tenm3 nanoclusters in WT synapses in the proximal CA1 region. i Summary graph of the average number of nanoclusters per Bassoon-positive excitatory synapse in Tenm3 WT and neuron cKO from proximal CA1. j The minimum intercluster distance between Bassoon and Tenm3 clusters. k–l Same as i, j, except that brain sections were analyzed from distal subiculum. Bar graphs show means ± SEM; numbers of sections/mice are indicated in the bars. Statistical significance was assessed using two-sided Student’s t-test (*p < 0.05; **p < 0.01). For additional experiments and quantifications, see Supplementary Fig. 2. Scale bars: 10 µm (a, d, left panel), 1 µm (a, d, right panel) and 200 nm (c, f).