Fig. 2.

Astrocyte-specific overexpression of HDAC7 leads to inflammation and anxiety-like behaviors in mice. AAV-GfaABC1D-HDAC7-eGFP or the empty vector was stereotaxically infused into the hippocampal CA1 subset of 2-month-old wildtype mice. After expression for 1-month, behavioral tests were performed. A Representative confocal image showing the co-expression of GFP, HDAC7, and GFAP in the dorsal hippocampal CA1 subset of 2-month-old mice. Scale bar, 50 μm. Quantification shows the relative GFAP expression level in GFP positive cells. N = 6 mice for each group, unpaired Student’s t test. B Astrocytic overexpression of HDAC7 increases mRNA levels of Il-1α, Il-1β, Il6, Tnfα, Gfap, and C3 analyzed by RT-qPCR in the hippocampus. N = 6 mice for each group, unpaired Student’s t test. C–K Astrocytic overexpression of HDAC7 activates NF-κB activity. C Representative western blots of HDAC7, GFAP, p-IKKα/β(S176/180), IKKα, IKKβ, p-NF-κB(S468), p-NF-κB(S536), NF-κB, and β-actin in the HDAC7-overexpressing and control hippocampal CA1 lysates. Quantification analysis of HDAC7 protein levels normalized to β-actin (D), GFAP protein levels normalized to β-actin (E), IKKα protein levels normalized to β-actin (F), IKKβ protein levels normalized to β-actin (G), p-IKKα/β(S176/180) normalized to IKKα (H), p-IKKα/β(S176/180) normalized to IKKβ (I), p-NF-κB(S468) normalized to total NF-κB (J), and p-NF-κB(S536) normalized to total NF-κB (K). N = 5 mice for each group, unpaired Student’s t test. L Representative immunofluorescence image showing enhanced nuclear NF-κB expression in HDAC7-overexpressing and control astrocytes. Scale bar, 10 μm. M Quantification of nuclear NF-κB fraction in GFP positive cells. N = 21–24 cells from 3 mice for each group, unpaired Student’s t test. N–R Overexpression of HDAC7 results in anxiety-like behaviors in the open field test. In this paradigm, the exploring performance in the center region (green frame area) is tested (N, left half panel). HDAC7-overexpressing mice show decreased center moving distance (O), center exploring time (P), and center entries (Q) in the open field test. The total moving distance remain unchanged (R). N = 8–11 mice, unpaired Student’s t test. N and S–U Overexpression of HDAC7 results in anxiety-like behaviors in the elevated plus maze. In this paradigm, the exploring performance in the open arm (green frame area) is tested (N, right half panel). HDAC7 mice show decreased open arm travel distance (S), open arm exploring time (T), and open arm entries (U) in the elevated plus maze. N = 8–11 mice, unpaired Student’s t test. Data were expressed as mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001