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

Fig. 2. Pathway-specific nucleus accumbens (NAc) astrocyte activity in response to medial prefrontal cortex (mPFC) afferents.

Fig. 2

A Scheme and representative slices showing opsin expression (ChrimsonR-tdTom) after AAV injection into the mPFC (top; scale bar = 1 mm) and their tdTom-expressing afferents to the NAc (bottom; scale bar = 500 µm). B Quantification of mPFC afferents fluorescence (arb.u.) in accumbens core (AcbC; slashed bar) and accumbens shell (AcbSh; solid bar) (11 infections, 6 mice). Two-tailed unpaired t test, *P = 0.02. C Scheme of NAc neuron’s electrophysiological recordings in response to mPFC afferent’s optostimulation. D Positive correlation between NAc neuron’s response (pA) and mPFC afferents’ density (ΔF/F0 in the area of register) (20 cells, 3 mice). E EPSC amplitude (pA) quantification (left) from AcbC (slashed bar; 8 cells, 4 mice) and AcbSh neurons (solid bar; 15 cells, 4 mice) and representative EPSCs traces (right) showing the synaptic neuronal strength triggered by mPFC in each subregion. Two-tailed unpaired t test Welch-corrected; **P = 0.02. F Scheme of astrocytic Ca2+ dynamics, monitored by real-time imaging of CaMPARIGFAP green fluorescence, in response to mPFC afferent’s optostimulation. Representative NAc slice (right) showing ChrimsonR-tdTom mPFC afferents (red) and CaMPARIGFAP astrocytes (green). Scale bar = 500 µm. G Heatmaps showing Ca2+ activity vs time (left; 150 ROIs, 3 slices) and representative Ca2+ traces (right), in response to mPFC optostimulation. H Proportion of responding ROIs (left) and relative Ca2+ spike frequency (right) induced by mPFC afferent’s optostimulation, at AcbC (slashed bar; 25 slices, 8 mice) and AcbSh (solid bar; 19 slices, 8 mice). Subsets of those ROIs were registered in the presence of the selective mGluR5 antagonist (MPEP, 50 µM) (magenta bars; AcbC and AcbSh pooled together, 4 slices, 2 mice). One-sample t test, #P < 0.05; ##P < 0.01; one-way ANOVA, Holm–Sidak test for multiple comparisons, P > 0.05. For more statistical detail, see Supplementary Table 1. Error bars express SEM. Source data are provided as a Source Data file.