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. 2022 Sep 7;13:5272. doi: 10.1038/s41467-022-33020-6

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© The Author(s) 2022

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. 7 — A Left, scheme of opsin-transfected vHip afferents in the NAc. Right, average PRQ image showing vHip glutamatergic innervation pattern. Orange lines starting from pixel 0 in each subregion were used for quantification (pixel = 50 µm²). B Left, glutamate afferents fluorescence (arb.u.) vs distance (pixels) quantifying vHip innervation across orange lines. Right, average glutamate afferents spatial fluorescence (arb.u.) in the accumbens core (AcbC; slashed bar) and accumbens shell (AcbSh; solid bar) (8 slices, 6 mice). One-sample t test, ^###P < 0.001; two-tailed unpaired t test, ***P < 0.001. C Left, scheme of astrocytic Ca²⁺ activity, measured by CaMPARI_GFAP red, in response to vHip afferent’s optostimulation. Right, average PRQ image showing astrocytic activation pattern in the NAc in basal and optostimulated conditions. Yellow lines starting from pixel 0 in each subregion were used for quantification (pixel = 50 µm²). D Left, CaMPARI_Red fluorescence (arb.u.) vs distance (pixels) quantifying astrocytic activation across yellow lines. Right, relative average CaMPARI_Red spatial fluorescence (arb.u.) in optostimulated condition in control, at AcbC (slashed bar) and AcbSh (solid bar) (8 pairs basal-stim slices, 6 mice), and in presence of MPEP (magenta bar; AcbC and AcbSh pooled together, 4 pairs basal-stim slices, 2 mice). One-sample t test, ^##P < 0.001; one-way ANOVA, Holm–Sidak test for multiple comparisons, *P = 0.02; **P = 0.002. E Masks of vHip glutamatergic afferents (red) and astrocyte activation area (yellow) defined by a k-mean clustering. In orange, the overlap area between the two. F Left, area (% from NAc) quantification of the spatial overlap (orange bar) between vHip afferents (red bar; P = 0.12) and active astrocytes (blue bar; **P = 0.003) (8 slices, 6 mice). One-way ANOVA, Holm–Sidak test for multiple comparisons. Right, bivariance index showing overlap distributions of AcbC and AcbSh. Note that AcbSh overlap area is embedded within the afferent’s area (Overlap/Glut. Affer. close to 1), indicating a direct interaction between vHip axons and astrocytes not present in AcbC. MANOVA test. Error bars express SEM. Source data are provided as a Source Data file.