Fig. 3: Inverting the natural transcriptional control of ZFP189 in the PFC with HSV-ZFP189^VPR impairs social function in mice.

A Experimental timeline for viral delivery, micro-defeat, social interaction, and elevated plus maze testing. B Mice treated with HSV-ZFP189^VPR spend less time interacting with a CD1 social target mouse. Social interaction (SI) ratio quantifying the time test mice spent in interaction zone when social target mouse cage is empty relative to when the cage is occupied with novel CD1 mouse. Mice treated with HSV-GFP, HSV-ZFP189^NFD, or HSV-ZFP189^WT prefer to socialize (SI ratio > 1), whereas HSV-ZFP189^VPR mice do not (SI ratio < 1). One-way ANOVA followed by Bonferroni’s multiple comparisons test. *p-value < 0.05; n = 21 mice (HSV-GFP) n = 18 mice (HSV-ZFP189^NFD), n = 25 mice (HSV-ZFP189^WT), n = 28 mice (HSV-ZFP189^VPR). C No viral treatment condition affected locomotor behaviors in the SI test, measured in the first 2.5 min session when the target CD1 was absent. D No viral treatment condition affected time spent on the open arm of the elevated plus maze. One-way ANOVA followed by Bonferroni’s multiple comparisons test. p-value > 0.05. E Experimental timeline for three-chamber social interaction test. F In session II, when given the choice between interacting with an empty cage or a novel mouse, HSV-GFP, HSV-ZFP189^NFD, and HSV-ZFP189^WT treated mice spend significantly more time interacting with the novel mouse, whereas HSV-ZFP189^VPR treated mice do not. Unpaired two-tailed Student’s t-test. ****p-value < 0.0001, *p-value < 0.05, ns p-value > 0.05. G Subjecting these same animals to session III, wherein test mice are given a choice between interacting with the caged mouse from the previous round (Familiar) or a novel mouse, HSV-GFP, HSV-ZFP189^NFD, and HSV-ZFP189^WT treated mice again spend more time interacting with the novel mouse, whereas HSV-ZFP189^VPR treated mice do not. *p-value < 0.05, ns p-value > 0.05. n = 18 mice (HSV-GFP), n = 13 mice (HSV-ZFP189^NFD), n = 10 mice (HSV-ZFP189^WT), n = 11 mice (HSV-ZFP189^VPR). H Experimental timeline for tube test. For five days, social hierarchy was determined by once daily head-to-head tube tests for all animals within a five-mouse cage. In each cage, either the two most socially dominant or the two most subordinate mice were delivered HSV-ZFP189 TFs intra-PFC whereas the remaining cage-mates were delivered HSV-GFP. Mice recovered for one day. For the following five days, the tube tests were repeated and wins for the HSV-ZFP189 TF treated mice versus all HSV-GFP treated cage-mates were recorded. I Delivering HSV-ZFP189^VPR to the PFC of previously socially dominant mice decreases the probability of winning a tube test to a rate of random chance (dotted line). Two-way repeated-measures ANOVA comparing main effect of wins pre- vs. post-viral intervention. ***p-value < 0.0001. n = 4 cages with two HSV-ZFP189^VPR mice per cage. J Delivering HSV-ZFP189^VPR to the PFC of previously socially subordinate mice increases the number of wins per day to a 50% chance of winning against an opponent. Two-way repeated-measures ANOVA comparing main effect of wins pre- vs. post-viral intervention. **p-value < 0.005. n = 6 cages with two HSV-ZFP189^VPR mice per cage. K Delivering HSV-ZFP189^NFD to the PFC of socially dominant mice does not change their tube test wins from pre-surgery performance. Two-way repeated-measures ANOVA comparing main effect of wins pre- vs. post-viral intervention. ns p-value > 0.05. n = 4 cages with two HSV-ZFP189^NFD mice per cage. L Delivering HSV-ZFP189^NFD to the PFC of socially subordinate mice does not change their tube test wins from pre-surgery performance. Two-way repeated-measures ANOVA comparing main effect of wins pre- vs. post-viral intervention. ns p-value > 0.05. n = 6 cages with two HSV-ZFP189^NFD mice per cage.