Article Figures & Data
Figures
- Figure 1.
Cdk5-ZFPs bidirectionally regulate Cdk5 expression and modify histones in vivo. A, Cdk5-ZFP1 and Cdk5-ZFP2 target the murine Cdk5 promoter at unique 18 bp recognition sites. The TSS and motifs for transcription factors NFIL6 and AP2 are also shown. B, qRT-PCR results show that Cdk5-ZFP1 fused to the transcriptional activation domain p65 increases Cdk5 mRNA expression when transfected into N2a cells, while Cdk5-ZFP2-p65 has no effect. N2a data are normalized to mock-transfected cells (n = 4,4,4). HSV-mediated delivery of Cdk5-ZFP1-p65 to NAc also increases Cdk5 mRNA expression in vivo. NAc data are normalized to HSV-p65, which lacks a DNA binding domain (n = 6,5). C, Cdk5-ZFP1 and Cdk5-ZFP2 fused to the transcriptional repression domain G9a have no effect on Cdk5 mRNA expression in transfected N2a cells (n = 4,4,4), while HSV-mediated delivery of Cdk5-ZFP2-G9a to NAc represses Cdk5 expression in vivo (n = 5). NAc data are normalized to HSV-G9a, which lacks a DNA binding domain (n = 4). D, qChIP for H3K9/14ac shows that HSV-Cdk5-ZFP1-p65 enriches histone acetylation at the Cdk5 promoter in NAc (n = 9,7). E, qChIP for H3K9me2 shows that HSV-Cdk5-ZFP2-G9a enriches histone methylation at the Cdk5 promoter in NAc (n = 5,5). F, qRT-PCR shows no regulation of putative off-target genes in NAc infected with either Cdk5-ZFP1-p65 or Cdk5-ZFP2-G9a (n = 5 ZFP, 5 control for each gene).
- Figure 2.
Epigenetic remodeling by Cdk5-ZFPs bidirectionally regulates cocaine (Coc) locomotor behavior. A, Mice were injected intra-NAc with HSVs expressing Cdk5-ZFP1-p65 (n = 10), Cdk5-ZFP2-G9a (n = 8), or control (Ctrl) viruses Cdk5-p65 (n = 9) or -G9a (n = 10), and were allowed to recover for 72 h before being subject to cocaine treatment for 4 d, followed by 8 d of abstinence and a challenge dose. B, At a moderate dose of cocaine (10 mg/kg, i.p.), Cdk5-ZFP1-p65 enhanced the locomotor-activating effects of cocaine over the 4 d course, with no difference observed after a challenge dose. C, At a higher dose of cocaine (15 mg/kg, i.p.), Cdk5-ZFP2-G9a diminished the locomotor effects of cocaine over the 4 d course, with no difference observed after a challenge dose. Neither virus had an effect on locomotor behavior following saline (Sal) treatment.
- Figure 3.
Epigenetic remodeling by Cdk5-ZFP2-G9a represses cocaine (Coc) place conditioning. A, Mice were injected intra-NAc with HSVs expressing Cdk5-ZFP1-p65 (n = 7), Cdk5-ZFP2-G9a (n = 6), or control GFP (n = 6), and were allowed to recover for 72 h before being subjected to CPP. B, At a moderate dose of cocaine (7.5 mg/kg, i.p.), control mice formed a significant preference for the cocaine-paired chamber, which was completely blocked by the injection of HSV-Cdk5-ZFP2-G9a. Injection of HSV-Cdk5-ZFP1-p65 had no effect on cocaine preference. C, At a low dose of cocaine (3.75 mg/kg, i.p.), neither control mice nor mice injected with HSV-Cdk5-ZFP1-p65 showed a preference for the cocaine-paired chamber, ruling out the possibility of a ceiling effect masking an effect of HSV-Cdk5-ZFP1-p65. Sal, Saline.
- Figure 4.
Epigenetic remodeling by Cdk5-ZFP1-p65 promotes resilience to social defeat stress. A, Mice were injected intra-NAc with HSVs expressing Cdk5-ZFP1-p65 (n = 9), Cdk5-ZFP2-G9a (n = 10), or control GFP (n = 9), and allowed to recover for 48 h before being subject to an accelerated social-defeat paradigm. Following 4 d of social defeat stress, mice were examined in a social interaction (SI) test, as well as EPM and sucrose preference tests. B, C, Social interaction tests revealed that control mice subjected to accelerated social defeat spent 25% less time in the interaction zone (B) and 28% more time in the corner zone (C) when a novel mouse was present, indicating a stress-susceptible phenotype, while mice injected with HSV-Cdk5-ZFP1-p65 did not show this social avoidance, indicating stress resilience. Mice injected with HSV-Cdk5-ZFP2-G9a behaved similarly to control mice, spending 19% less time in the interaction zone and 18% more time in the corner zone when a novel mouse was present.
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