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. 2016 Jul 12;22(11):1576–1584. doi: 10.1038/mp.2016.103

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Copyright © 2016 Macmillan Publishers Limited

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/

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Increased dendritic spine density in mice with Atg7-deficient microglia. (a) Representative images of dendrites in the somatosensory 2 (S2) brain region from Atg7^fl/fl and Atg7^fl/fl;Lyz2-Cre mouse after Golgi-Cox staining. The number of dendritic spines in Atg7^fl/fl;Lyz2-Cre mice was significantly increased compared with control mice at postnatal day 15. n=150 segments for Atg7^fl/fl, n=60 segments for Atg7^fl/fl;Lyz2-Cre. Scale bar=2 μm. (b) PSD95 and synaptophysin immunoreactivity was more frequently co-localized with Iba-1 immunoreactivity in the brain of Atg7^fl/fl;Lyz2-Cre mice than that of Atg7^fl/fl mice. Representative images from each group are shown. Scale bar=20 μm. (c) Three-dimensional confocal image, constructed from serial confocal z-stack images, of Iba-1-positive microglia (red) from an Atg7^fl/fl;Lyz2-Cre mouse. High-magnitude images of PSD95-positive puncta (green) engulfed by Iba-1-positive microglia are shown in the lower panel. The number of PSD95-positive puncta per microglia (yellow) was significantly increased in Atg7^fl/fl;Lyz2-Cre mice compared with controls at postnatal day 12. n=40 cells for each group. (d) Western blot images of synaptic markers (PSD95 and SHANK3) and autophagy-related proteins (LC3-II and p62). Intensities of both PSD95 and SHANK3 were significantly higher in Atg7^fl/fl;Lyz2-Cre mice than in the controls. In Atg7^fl/fl;Lyz2-Cre mice, the LC3-II intensity was decreased and that of p62 is increased compared with Atg7^fl/fl mice. n=5 for each group. *P<0.05, **P<0.01, ***P<0.001 (two-tailed Student’s t-test).