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

Fig. 5. Spatial nucleus accumbens (NAc) astrocytic response to basolateral amygdala (Amyg) afferents.

Fig. 5

A Left, scheme of opsin-transfected Amyg afferents in the NAc. Right, average PRQ image showing Amyg 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 Amyg 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 = 0.017, ##P = 0.0021; two-tailed unpaired t test, *P = 0.04. C Left, scheme of astrocytic Ca2+ activity, measured by CaMPARIGFAP red, in response to Amyg 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 condition vs basal values 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 = 0.0261, ##P = 0.0054; One-way ANOVA, Holm–Sidak test for multiple comparisons, **P = 0.007. E Masks of Amyg 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 Amyg afferents (red bar; **P = 0.001) and active astrocytes (green bar; **P = 0.003) (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 Amyg afferents. Note that there is no difference between subregions. MANOVA, d = 0, P = 0.482. Error bars express SEM. Source data are provided as a Source Data file.