Fig. 6

Rescue of Negr1 expression ameliorates behavioral and synaptic deficits induced by the gain of AP1AR-DT in mice. A–E rAAVs carrying lncRNA-AP1AR-DT (OE), Negr1(for rescue of Negr1 expression, RE), or empty rAAV vectors (Ctl) were cloned into the rAAV expression vector and injected into the mPFC regions of C57BL/6 wild-type mice (A), and their expression levels were examined by immunofluorescence analysis of brain slices (B) or by qRT-PCR analysis of AP1AR-DT (C) and the Negr1 mRNA (D) levels and Western blotting analysis of Negr1 protein levels using an anti-mCherry antibody for recombinant Negr1 and an anti-Negr1 antibody for endogenous Negr1(E). The time line for each behavior test is also indicated. F The reduced spine densities in AP1AR-DT OE mice (OE) neurons were rescued in Negr1 rescued mice (RE). n = 35 neurons from Ctl mice, n = 36 neurons from OE mice, and n = 48 neurons from RE mice. G–K Rescue of Negr1 expression in AP1AR-DT OE mice restored the immobile times of TST (n = 15 Ctl mice, n = 15 OE mice, and n = 13 RE mice, G) and FST (n = 15 Ctl mice, n = 15 OE mice, and n = 15 RE mice, H), time spent in the central area of the OFT (n = 13 Ctl mice, n = 13 OE mice, and n = 14 RE mice, I–J), and time spent in the open arms of the EPM (n = 11 Ctl mice, n = 11 OE mice, and n = 11 RE mice, K). L Rescue of Negr1 expression in AP1AR-DT OE mice ameliorates the impaired sEPSCs induced by AP1AR-DT_OE. Representative recording traces and cumulative probability plots of sEPSCs showing that the decreased sEPSC frequency in AP1AR-DT_OE mice (OE) was increased when the Negr1 expression was restored (RE) in mPFC neurons. n = 13 cells from four Ctl mice, n = 18 cells from three OE mice, and n = 12 cells from three RE mice. All data represent the means ± SEMs. A two-tailed t-test was used for comparisons between the two indicated groups (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. ns, nonsignificant)