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. 2020 Sep 29;13:149. doi: 10.3389/fnmol.2020.00149

Figure 1.

Figure 1

VGLUT1 inclusions can be found in adult hippocampal microglia under homeostasis. (A) In immunohistochemistry, frequent overlaps between Iba1 and VGLUT1 signals can be detected in the brain cortex and dentate gyrus (DG). (B) Anti-VGLUT1 staining in cortex and dentate gyrus (DG) from both Vglut1+/+ (wild-type) and Vglut1−/− (knock-out) littermates. (C) Quantification of the VGLUT1-positive voxels per stack in cortex (CTX) and dentate gyrus (DG) of both Vglut1+/+ and Vglut1−/− mice (N = 3 mice per group, two sections per mouse; data from a single experiment). (D) Microglial cell at the CA1 hippocampus exhibiting a VGLUT1 inclusion overlapping with a process (yellow arrowhead). (E) 3D cell reconstruction of the same microglial cell displayed in panel (D). (F) Application of a clipping surface on the 3D reconstruction discloses the presence of a VGLUT1 inclusion within the microglial process (yellow arrowhead). (G) STED image of a microglial cell in the CA1 exhibiting a VGLUT1 inclusion (white arrowhead). (H) Orthogonal projection of the imaged in panel (J) showing the intracellular localization of the VGLUT1 staining. (I) 3D reconstruction of the cell in panel (J) showing the intracellular localization of the VGLUT1 staining. (J) A confocal image of a microglial cell body exhibiting VGLUT1 inclusion overlapping with a CD68+ phagosome (scale bar 5 μm). (K) Orthogonal projection of the inset area. Note the colocalization between VGLUT1 and CD68 staining (white spot in the centre of the crosshair). (L) 3D reconstruction of both CD68 (green) and VGLUT1 (blue) signals showing that the VGLUT1 inclusion is perfectly encased within the microglial phagosomes (white arrowhead).