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. 2022 Aug 11;14:968404. doi: 10.3389/fnsyn.2022.968404

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Copyright © 2022 Gore, Yurina, Yukevich-Mussomeli and Nahmani.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Clathrin-coated (CT) and uncoated spinules (no-CT) are distinct populations within CA1 SBBs. (A) Cartoon showing the percentage of SBBs containing at least one spinule from adjacent axons/boutons (AdjAx), postsynaptic spines (PSs), adjacent (no synapse with SBB) spines (AdjS), adjacent dendrites (AdjD), glia, and postsynaptic dendrites (PSd). Percentages reflect the ∼42% of SBBs with spinules from multiple sources. (B) Percentage of perforated PSDs in postsynaptic spines in this image volume. Upper left panel shows a FIB-SEM image of a postsynaptic spine (pink) with a perforated PSD (red) and its presynaptic SBB partner (purple). Note the CT spinule projecting just to the right of the PSD. Upper right panel shows 3D reconstruction of spine and PSD from FIB-SEM image; note this is the same spine as shown in Figures 1G₂,G₃. Lower left panel shows the percentages of spinule-bearing (SB) and non-spinule-bearing (non-SB) postsynaptic spines displaying perforated PSDs. Lower right panels show the percentages of spines projecting PSs spinules and AdjS spinules into SBBs that display perforated PSDs. (C) Percentage of spines with a spine apparatus in this image volume. Upper left shows a FIB-SEM image of the same postsynaptic spine (outlined in pink) as in panel (B), displaying a spine apparatus (cyan), with a synapse onto an SBB (purple). Upper right panel shows the 3D reconstruction of this spine and its spine apparatus. Lower left panel shows the percentages of SB and non-SB spines displaying a spine apparatus, while lower right panels show the percentages of spines projecting PSs and AdjS spinules containing a spine apparatus. (D) Spinule volume comparisons between no-CT (blue) and CT spinules (pink) (D₁), between CT spinules of AdjAx (green), AdjS (yellow), and PSs origins (magenta) (D₂), and between no-CT spinules of AdjAx, AdjS, and PSs origins (D₃). For all boxplots, center lines show medians, box limits indicate the 25th and 75th percentiles, whiskers extend to the 5th and 95th percentiles, width of boxes is proportional to the square root of the sample size, and raw data points are plotted as filled circles. (E) Comparisons of spinule penetration distance into their SBBs between no-CT and CT spinules (E₁), between CT spinules of AdjAx, AdjS, and PSs origins (E₂), and between no-CT spinules of AdjAx, AdjS, and PSs origins (E₃). (F) Comparisons of the percentage of SBB volume occupied by no-CT and CT spinules (F₁), CT spinules of AdjAx, AdjS, and PSs origins (F₂), and no-CT spinules of AdjAx, AdjS, and PSs origins (F₃). Scale bars (B,C) 0.5 μm; Scale cubes (B,C) 0.5 μm per side.