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

Fig. 3. Spatial nucleus accumbens (NAc) astrocytic response to medial prefrontal cortex (mPFC) afferents.

Fig. 3

A Left, scheme of opsin-transfected mPFC afferents in the NAc. Right, average PRQ image showing mPFC glutamatergic innervation pattern. Orange lines starting from pixel 0 in each subregion were used for quantification (pixel = 50 µm2). B Left, Glut. afferents fluorescence (arb.u.) vs distance (pixels) quantifying mPFC innervation across orange lines. Right, average glutamatergic afferents spatial fluorescence (arb.u.) in the accumbens core (AcbC; slashed bar) and accumbens shell (AcbSh; solid bar) (9 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 CaMPARIGFAP red, in response to mPFC 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, CaMPARIRed fluorescence (arb.u.) vs distance (pixels) quantifying astrocytic activation across yellow lines. Right, average CaMPARIRed spatial fluorescence (arb.u.) in optostimulated slices in control, at AcbC (slashed bar) and AcbSh (solid bar) (9 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 < 0.01; one-way ANOVA, Holm–Sidak test for multiple comparisons, *P < 0.05, **P < 0.01. For more statistic detail, see Supplementary Table 1. E Masks of mPFC 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 mPFC afferents (red bar; *P = 0.04) and active astrocytes (black bar; **P = 0.001) (9 slices, 6 mice). One-way ANOVA, Holm–Sidak test for multiple comparisons. Right, bivariance index showing overlap distributions of AcbC and AcbSh; indicates astrocytes interaction with mPFC afferents. Note that there is no difference between subregions. MANOVA, d = 0, P = 0.586. Error bars express SEM. Source data are provided as a Source Data file.