Article Figures & Data
Figures
- Figure 1.
Gene-specific changes in hippocampal H2A.Z and AcH2A.Z binding to memory-related genes 24 h after fear conditioning. a, b, H2A.Z (red; a) and AcH2A.Z (blue; b) binding upstream (−1 nucleosome) and downstream (+1 nucleosome) from the TSS. Data are expressed as the mean ± SEM for each group. N mice, n = 10; C mice, n = 9; S mice, n = 10; CS mice, n = 10. *p ≤ 0.05.
- Figure 2.
Hippocampal H3 binding at memory-related genes 24 h after fear conditioning. H3 binding is expressed as the mean ± SEM for each group normalized to naive controls. For each gene, data are shown separately for sites upstream (−1 nucleosome) and downstream (+1 nucleosome) of the TSS.
- Figure 3.
Gene-specific changes in cortical H2A.Z and AcH2A.Z binding to memory-related genes 30 d after fear conditioning. a, b, H2A.Z (a) and AcH2A.Z (b) binding in the cortex is shown for sites upstream (−1 nucleosome) and downstream (+1 nucleosome) of the TSS. Data are expressed as the mean ± SEM. N mice, n = 13–15; C mice, n = 6–10; S mice, N = 8–9; CS mice, n = 10–11. *p ≤ 0.05.
- Figure 4.
Cortical histone H3 binding at memory-related genes 30 d after fear conditioning. H3 binding is expressed as the mean ± SEM for each group normalized to naive controls. For each gene, data are shown separately for sites upstream (−1 nucleosome) and downstream (+1 nucleosome) of the TSS.
- Figure 5.
Blocking Tip60 via Nu9056 inhibits H2A.Z expression and binding. a, Outline of the Nu9056 treatment protocol in cultured hippocampal neurons. b, Representative image of immunoblotting for H2A.Z and acH2A.Z (left) and protein quantification (right). Dashed line represents the average of DMSO-treated control neurons (n = 3 wells/group). c, Gene expression changes in response to Nu9056 treatment in cultured hippocampal neurons. Dashed line represents the average of DMSO-treated control neurons (n = 6 wells/group). d, H2A.Z (top) and AcH2A.Z (bottom) binding at −1 (left) and +1 (right) nucleosomes in neurons treated with Nu9056 (n = 6 wells/group). e, Outline of Nu9056 treatment protocol in mice. f, H2A.Z (top) and AcH2A.Z (bottom) binding at −1 (left) and +1 (right) nucleosomes in the hippocampus 1 h after Nu9056 injection (n = 7 mice/group). Data are expressed as the mean ± SEM relative to DMSO controls. *p ≤ 0.05. Veh, vehicle.
- Figure 6.
Blocking Tip60 has stage-specific effects on memory. a, Outline of experimental design. Fear memory is indexed as a percentage of the time spent freezing. b, The Tip60 inhibitor Nu9056 does not affect memory when tested 24 h after learning (i.e., 1 h after injection), but promotes memory when tested 7 d after learning (i.e., 6 d after injection) when the same mice are tested at both time points. c, Nu9056 improved remote memory in a separate group of mice that were not tested at 24 h (i.e., recall was tested for the first time 7 d after training/6 d after injection). Data are expressed as the mean ± SEM. Vehicle (Veh), N = 12; Nu9056, N = 11. *p ≤ 0.05.
- Figure 7.
TSA increases H2A.Z acetylation, but does not affect remote memory. a, b, Immunoblot showing levels of H2A.Z and AcH2A.Z after treatment with TSA in cultured hippocampal neurons (N = 3 wells/group; a) and adult mouse hippocampus after systemic injection of TSA (N = 4 mice/group; b). Separate actin is used for normalizing H2A.Z and acH2A.Z expression because each protein was run on a separate gel to facilitate visualization of similar size proteins. c, Outline of experimental design (top), and memory recall for recent (24 h) and remote (7 d) time points (n = 12/group). Data are expressed as the mean ± SEM. *p ≤ 0.05.
- Figure 8.
Blocking Tip60 has stage-specific effects on memory. Outline of experimental design is shown on top. Fear memory is indexed as the percentage of time spent freezing. Data are expressed as the mean ± SEM. Vehicle (Veh), N = 9; Nu9056, N = 10. *p ≤ 0.05.
Tables
Forward sequence Reverse sequence +1 Arc 5′-GCCACACTCGCTAAGCTCC 5′-AACTCCTCTGAGGCAGAAGCC −1 Arc 5′-TCCCGGTGGGAGGCG 5′-GTGCCCTCAAGGACCCG +1 Fos
−1 Fos
+1 Gria4
-1 Gria45′-AGTGTCTACCCCTGGACCC
5′-AGGAGACCCCCTAAGATCCC
5′-CCATGGGGAGGTGCCTAACTT
5′-TTCTAGACCCCAGCCTCATCA5′-GCGTTGAAACCCGAGAACATC
5′-CTGTCGTCAACTCTACGCCC
5′-GCTTGGCAGGAAACTACCCC
5′-CGGATGGATGCGGTAAAGGTA+1 Kcna2
-1 Kcna25′-GGATTAGACATCCAGCTCAGG
5-GATGTGCAAGCGAAGAACCC5′-CTCCCCCATCTCTCTTGGTA
5′-GAGATGGCAGCACTGGCT+1 Syt1
-1 Syt15′-GGGATGCTCTGACCGAGTTC
5′-TGGGATCTGGTCTCCTCTTAGT5′-CTGTTGGGTGAGCAGGACC
5′-GCTCCTCTCAGGCAACGGA+1 Syp
−1 Syp
+1 Tacstd2
-1 Tacstd25′-GACTGGGCTGTTCCGACGAT
5′-CAAACAGGACTCAGGTTGGC
5′-TCCTACCCAGCCTGATCCTTC
5′-GCAATCTCCCCTGCCTGATT5′-TTCACCACGTCCATGTCTGC
5′-TCCATTCGGGAGGCTAGAGA
5′-AGCGGTGCTAGATCCAAGCC
5′-TCAATGGAATTTGGGTGGCG
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