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. Author manuscript; available in PMC: 2018 Nov 2.
Published in final edited form as: Science. 2017 Sep 28;357(6358):1406–1411. doi: 10.1126/science.aan4994

Fig. 2. PVN OXT neuronal activity is enhanced during social interaction and is sufficient to induce social real-time CPP.

Fig. 2.

(A) Ai14 tdTomato reporter mice were injected with CAV2-Cre in the VTA and subjected to interaction with either a juvenile mouse (n = 5) or a toy mouse (n = 3). (B) Quantification of the percentage of tdTomato-positive cells, whichare known to project to the VTA, that were also c-fos–positive. (C and D) Diagrams showing the methods used to record GCaMP6m activity in PVN OXT neurons. (E) Immunohistological image showing fiber placement in close proximity to PVN OXT neurons (red) expressing GCaMP6m (green). Scale bar,100 μm. (F and G) Time course plots showing event-locked average z-scores of GCaMP6m transients in response to social interactions with a toy mouse (n = 16 events from an individual mouse) or with a juvenile mouse (n = 7 events from an individual mouse). Shaded areas, SEM. ΔF/F, normalized change in fluorescence. (H) Plot of average fluorescent signal for 1 s beforeand 1 s after the onset of a toy interaction event (n = 26 events from four mice) or juvenile interaction event (n = 25 events from six mice). (I) Diagram showing how AAVDJ-DIO-ChETA-eYFP was injected into the PVN of OXTiresCre mice with optical fibers implanted in the PVN. (J) Immunohistological images showing fiber placement and colocalization of eGFP (green) and OXT neurons (red) in the PVN. Scale bar, 100 μm. 3V, third ventricle. (K) Quantification of real-time CPP (RT-CPP) for eYFP-expressing (black; n = 4) and ChETA-expressing (red; n = 6) mice. (L) Operant nose-poking to receive 30-Hz light stimulation did not differ between eYFP- and ChETA-expressing mice (n = 8 for both groups). (M) Diagram showing the social RT-CPP (sRT-CPP) protocol. (N and O) Time spent in social chamber on day 1 (D1) subtracted from time spent on day 2 (D2) for eYFP- and ChETA-expressing mice, graphed in 5-min bins (n = 10 mice in each group). (P) Cumulative subtracted preference over the 15-min test period. Data shown are means ± SEM. Significance was calculated by means of unpaired t tests for across-group comparisons in (B), (H), and (K) and two-way analysis of variance (ANOVA) for (P). *P < 0.05; **P < 0.01; ***P < 0.001.