1. Volume 97, Issue 5, Pages 1137-1152.e5 (March 2018)
Npas4 Is a Critical Regulator of Learning-Induced Plasticity at Mossy Fiber-CA3 Synapses during Contextual Memory Formation 
Feng-Ju Weng, Rodrigo I. Garcia, Stefano Lutzu, Karina Alviña, Yuxiang Zhang, Margaret Dushko, Taeyun Ku, Khaled Zemoura, David Rich, Dario Garcia-Dominguez, Matthew Hung, Tushar D. Yelhekar, Andreas Toft Sørensen, Weifeng Xu, Kwanghun Chung, Pablo E. Castillo, Yingxi Lin 
Neuron 
Volume 97, Issue 5, Pages e5 (March 2018)
DOI: /j.neuron
Copyright © 2018 Elsevier Inc. Terms and Conditions

2. Figure 1
Excitatory Miniature Activity in CA3 but Not CA1 Pyramidal Neurons Is Significantly Increased in Npas4 KO Mice, whereas Inhibitory Miniature Activity Is Not Affected
(A) Sample traces (top left, raw traces; top right, average mEPSCs), summary bar graphs, and cumulative distribution of mEPSCs recorded from hippocampal CA3 pyramidal neurons. Frequency: wild-type (WT), 5.95 ± 0.71 Hz; KO, 11.05 ± 1.72 Hz; p = Amplitude: WT, 16.61 ± 0.70 pA; KO, 22.12 ± 1.59 pA; p =  WT: n = 36 cells, 6 animals; KO: n = 34 cells, 5 animals.
(B) The same as (A) but for CA1 pyramidal neurons. Frequency: WT, 2.70 ± 1.09 Hz; KO, 2.01 ± 0.38 Hz; p = Amplitude: WT, 10.82 ± 0.82 pA; KO, 11.72 ± 0.35 pA; p = WT, n = 10 cells, 3 animals; KO, n = 16 cells, 3 animals.
(C) Sample traces, summary bar graphs, and cumulative distribution of mIPSC recorded from hippocampal CA3 neurons. Frequency: WT, 7.04 ± 0.77 Hz; KO, 8.16 ± 1.09 Hz; p = Amplitude: WT, 33.91 ± 2.10 pA; KO, 37.14 ± 1.67 pA; p = WT: n = 22 cells, 3 animals; KO: n = 22 cells, 4 animals.
(D) The same as (C) but for CA1 pyramidal neurons. Frequency: WT, 5.41 ± 1.23 Hz; KO, 5.47 ± 1.46 Hz; p = Amplitude: WT, 26.24 ± 2.13 pA; KO, 28.92 ± 1.32 pA; p = WT: n = 10 cells, 3 animals; KO: n = 11 cells, 3 animals.
Data are shown as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, Mann-Whitney U test.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions

3. Figure 2
The mEPSC Frequency in CA3 Pyramidal Neurons Is Significantly Altered by Acute Manipulation of Npas4 Expression
(A) AAV1-hSyn-Cre-EGFP (Cre) or AAV1-hSyn-EGFP (GFP) was stereotaxically injected into the CA3 or CA1 region of Npas4flx/flx mice at 8–10 weeks of age. Whole-cell voltage-clamp recordings were performed 3 days after virus injection.
(B) Representative images showing complete Npas4 gene deletion from the CA3 (top) or CA1 (bottom) region in Npas4flx/flx mice 3 days after injection of Cre. Npas4 expression (red) following kainic acid-induced seizure was completely absent in regions expressing Cre (green). The scale bars correspond to 300 μm.
(C) Sample traces, summary bar graphs, and cumulative distribution of mEPSCs recorded from hippocampal CA3 pyramidal neurons from Npas4flx/flx mice injected with Cre or GFP. Frequency: GFP, 6.55 ± 0.75 Hz; Cre, 12.61 ± 1.26 Hz; p = Amplitude: GFP, 16.22 ± 0.73 pA; Cre, 17.83 ± 0.74 pA; p = GFP: n = 32 cells, 6 animals; Cre: n = 28 cells, 6 animals.
(D) The same as (C) but for CA1 pyramidal neurons. Frequency: GFP, 2.53 ± 0.30 Hz; Cre, 3.10 ± 0.52 Hz; p = Amplitude: GFP, 10.19 ± 0.32 pA; Cre, 10.09 ± 0.32 pA; p = GFP: n = 16 cells, 4 animals; Cre: n = 15 cells, 3 animals.
(E) HSV-EGFP, HSV-Npas4-EGFP, or HSV-ΔNpas4-EGFP was stereotaxically injected into the CA3 region of Npas4 KO mice at 8–10 weeks of age. Whole-cell voltage-clamp recordings were performed 3 days after virus injection.
(F) Representative images showing acute re-expression of Npas4 in CA3 of Npas4 KO mice using HSV-Npas4-EGFP (bottom). HSV-EGFP was used as a control (top). The scale bars correspond to 300 μm.
(G) Sample traces, cumulative distribution, and summary bar graphs of mEPSCs recorded from hippocampal CA3 pyramidal cells of Npas4 KO mice after expression of Npas4, GFP, or ΔNpas4 or from uninfected cells. Horizontal lines indicate the levels in WT animals (from Figure 1A). Frequency: Npas4, 4.68 ± 0.98 Hz; GFP, 14.94 ± 3.30 Hz; ΔNpas4, 12.05 ± 1.88 Hz; uninfected, 14.10 ± 2.16 Hz; F (3, 65) = 3.739; p = Amplitude: Npas4, 19.09 ± 1.43 pA; GFP, 21.75 ± 1.84 pA; ΔNpas4, 21.04 ± 1.43 pA; uninfected, 22.04 ± 1.36 pA; F (3, 65) = ; p = Npas4: n = 15 cells, 3 animals; GFP: n = 15 cells, 3 animals; ΔNpas4: n = 13 cells, 3 animals; uninfected: n = 26 cells, 9 animals.
Data are shown as mean ± SEM. In (C) and (D), ∗∗∗p < 0.001, Mann-Whitney U test. In (G), ∗p < 0.05, one-way ANOVA, Dunnett’s post hoc test. See also Figure S1.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions

4. Figure 3
Deletion of Npas4 in CA3 Selectively Alters the Excitatory Inputs Received by CA3 Pyramidal Neurons from MFs, whereas Short-Term and Long-Term Plasticity Remains Unchanged
(A and B) Npas4 is not required in CA3 for retrieval of contextual memory.
(A) Npas4 cKO mice underwent CFC training, and their contextual memories were recalled the following day (recall 1). Npas4 was deleted 1 day after recall 1, and contextual memories were re-examined 3 days after surgery (recall 2).
(B) All animals, regardless of the presence or absence of Npas4, displayed similar freezing levels during both recalls. GFP freezing level: recall 1, 64.58% ± 5.22%; recall 2, 56.25% ± 5.35%; p = 0.1. Cre freezing level: recall 1, 65.34% ± 6.85%; recall 2, 59.85% ± 5.94%; p = Wilcoxon matched pairs signed rank tests were used to compare recall 1 and recall 2 for each treatment.
(C) AAV1-hSyn-EGFP (GFP) or AAV1-hSyn-Cre-EGFP (Cre) was stereotaxically injected into the CA3 region of Npas4flx/flx mice at 8–10 weeks of age. Whole-cell voltage-clamp recordings were performed 3 days after virus injection. In the presence of 1 μM DCG-IV (to block MF transmission), Npas4 deletion had no effect on the frequency or amplitude of mEPSCs recorded from CA3 pyramidal neurons. Sample traces, summary bar graphs, and cumulative distributions are shown. Frequency: GFP, 8.60 ± 0.89 Hz; Cre, 7.68 ± 0.93 Hz; p = Amplitude: GFP, 17.10 ± 0.82 pA; Cre, 17.53 ± 0.83 pA; p = GFP: n = 28 cells, 7 animals; Cre: n = 24 cells, 7 animals.
(D) Representative experiments of MF-CA3 EPSCs elicited by minimal stimulation in WT and Npas4 KO mice. Shown are superimposed individual traces (top) and time course plots (bottom). Adding DCG-IV (1 μM) at the end of the experiments blocked synaptic transmission (traces not shown).
(E) Summary data showing efficacy: WT, 43.3 ± 8.9 pA; KO, 132.8 ± 25.7 pA; p = Potency: WT, 78 ± 13.4 pA; KO, 184 ± 34.1 pA; p = Failure rate: WT, 28.7% ± 4.6%; KO, 30.5% ± 4.8%; p = WT: 8 slices, 6 mice; KO: 6 slices, 4 mice.
(F) Short-term plasticity was not altered in Npas4 KO mice. Extracellular MF synaptic responses (fEPSPs) were recorded in acute hippocampal slices from Npas4 KO mice and their WT littermates at 8–10 weeks of age. Sample traces and summary plots are shown. Left: the PPR was assessed by delivering paired pulses at 10-, 30-, 100-, 300-, and 1,000-ms inter-stimulus intervals, and pulse (P)2/P1 are plotted. Right: frequency facilitation was probed by 5 consecutive pulses at 40-ms intervals, and Pn/P1 are plotted. WT: n = 7 slices, 3 animals; KO: n = 8 slices, 3 animals.
(G) LTP at MF-CA3 synapses was normal in Npas4 KO mice. LTP was induced with a train of 125 stimuli at 25 Hz. DCG-IV (1 μM) was added at the end of each experiment. Representative traces are shown on the top and summary plots on the bottom. LTP (percent of the baseline for the last 10 min): WT, % ± 2.52%, n = 7 slices, 3 animals; KO, % ± 2.01%, n = 8 slices, 3 animals; p = 0.55.
All summary data are shown as mean ± SEM. In (C)–(G), ∗∗p < 0.01, Mann-Whitney U test. n.s., non-significant. See also Figures S2 and S3.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions

5. Figure 4 Npas4 Modifies the Number and Structure of MF-CA3 Synapses
(A) Schematic of the labeling strategy with CA3 pyramidal cells expressing nuclearly localized Cre-GFP or ΔCre-GFP and cytosolic tdTomato to label morphology. MF projections from the DG are labeled with both cytosolic and membrane-bound farnesylated GFP.
(B) Representative low-resolution image of two-color labeling of CA3 pyramidal cells (red, tdTomato) and DG cells and their MF projections (green, GFP). Inset: enlarged representation of a CA3 pyramidal cell with nuclear expression of either Cre or ΔCre (green). Scale bar, 500 μm.
(C and D) Top: representative images of MF terminals expressing GFP in the stratum lucidum of CA3 under ΔCre (C) or Cre (D) conditions. Bottom: examples of 3D surface reconstruction of large single MF terminals. Scale bars, 10 μm and 1 μm.
(E and F) Representative images of TEs on a proximal CA3 dendrite segment (left) and partial 3D surface reconstruction of a dendritic segment (right) for ΔCre (E) and Cre (F) conditions. Scale bars, 5 μm.
(G) MF terminals that synapse onto Npas4-deleted cells have significantly increased volumes. Volume: ΔCre, 26.06 ± 1.56, n = 29 MF terminals, 3 animals; Cre, 32.48 ± 1.53, n = 33 MF terminals, 3 animals; p =
(H) The volume of TE structures is significantly increased in Npas4-deleted cells. Volume: ΔCre, 2.91 ± 0.25, n = 9, 3 animals; Cre, 3.85 ± 0.28, n = 9, 3 animals; p = 0.04.
(I) Representative image of MAP-processed sections with labeling of the synaptic markers PSD-95 (magenta) and Bassoon (cyan). The inset shows a 3D surface reconstruction of synaptic marker puncta in the region marked with a yellow box. The dashed line indicates the boundary between the stratum lucidum and radiatum in CA3. Scale bars, 10 μm and 1 μm.
(J) The density of pre- and post-synaptic marker puncta is significantly increased in the stratum lucidum of Npas4-deleted cells. Synaptic density: Bassoon, ΔCre 0.07 ± 0.004, n = 17 regions, 6 animals; Bassoon, Cre 0.09 ± 0.004, n = 17 regions, 6 animals; p = 0.008; PSD-95, ΔCre 0.06 ± 0.004, n = 17 regions, 6 animals; PSD-95, Cre 0.07 ± 0.005, n = 17 regions, 6 animals; p = 0.02.
(K) The density of co-localized Bassoon and PSD-95 puncta is significantly increased in the lucidum of Npas4-deleted mice. Synaptic density: Bassoon, ΔCre 0.05 ± 0.004, n = 17 regions, 6 animals; Cre 0.07 ± 0.006, n = 17 regions, 6 animals; p =
(L) The average synaptic punctum volume does not change when Npas4 is deleted. Average volume: Bassoon, ΔCre 0.05 ± 0.004, n = 12 regions, 6 animals; Cre 0.05 ± 0.001, n = 12 regions, 6 animals; p = 0.57; PSD-95, ΔCre 0.04 ± 0.003, n = 12 regions, 6 animals; PSD-95, Cre 0.04 ± 0.002, n = 12 regions, 6 animals; p = 0.42.
(M) The density of Bassoon puncta localized within MF terminals does not change in Npas4-deleted cells. Synaptic density: Bassoon, ΔCre 1.89 ± 0.31, n = 12 regions, 6 animals; Cre 1.89 ± 0.17, n = 12 regions, 6 animals; p = 0.98.
Summary data are shown as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, Mann-Whitney U test.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions

6. Figure 5 Npas4 Bi-directionally Regulates TE Size
(A) Representative images and summary bar graphs showing that the total area of CA3 TE is significantly increased when Npas4 is deleted by AAV-Cre in Npas4flx/flx mice compared with littermate controls injected with AAV-GFP. TE area: GFP, 0.89 ± 0.06 μm2/μm, n = 15, 3 animals; Cre, 1.21 ± 0.08 μm2/μm, n = 15, 3 animals; p =
(B) Representative images and summary bar graphs showing a significantly larger total area of CA3 TE in Npas4 KO mice than in their WT littermates. TE area: WT, 0.71 ± 0.07 μm2/μm, n = 15 neurons, 3 animals; KO, 1.18 ± 0.12 μm2/μm, n = 15 neurons, 3 animals; p =
(C) Representative images and summary bar graphs showing comparable spine density on distal dendrites of CA3 pyramidal neurons in Npas4 KO mice and their WT littermates. Number of basal spines per 10 μm: WT, 18.49 ± 0.64; KO, 19.25 ± 0.59; p = Number of apical spines per 10 μm: WT, 15.25 ± 0.58; KO, 14.84 ± 0.57; p = WT basal: n = 17 dendrites, 2 animals; KO basal: n = 23 dendrites, 3 animals; WT apical: n = 42 dendrites, 7 animals; KO apical: n = 49 dendrites, 8 animals.
(D) Representative images and summary bar graphs showing that Npas4 expression in Npas4 KO mice reduced CA3 TE size to the WT level. TE area: ΔNpas4, 1.25 ± 0.12 μm2/μm; Npas4, 0.86 ± 0.06 μm2/μm; n = 15 neurons, 3 animals for each condition; p = The blue and brown lines indicate the TE area values of Npas4 WT and KO mice, respectively.
Scale bars correspond to 5 μm. Summary data are shown as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, Mann-Whitney U test.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions

7. Figure 6
Contextual Learning Selectively Strengthens MF Inputs, and Npas4 Deletion Prevents This Experience-Dependent Modulation
(A) The design of the RAM system. The RAM promoter PRAM drives the expression of a destabilized version of the tetracycline transactivator (d2tTA), which subsequently drives the expression of the reporter gene under the control of a tTA-responsive element (TRE) in the promoter. The binding of d2tTA to TRE and, as a result, the expression of the reporter gene can be blocked by the antibiotic doxycycline (Dox).
(B) Experimental timeline. Animals were taken off the Dox diet 5 days after virus injection and 48 hr before CFC training and sacrificed 24 hr later for electrophysiological recordings.
(C) WT C57BL/6 mice were co-injected with AAV-RAM-mKate2 and AAV-EF1α-GFP (as an infection control). RAM+ CA3 pyramidal neurons have a significantly higher mEPSC frequency than their neighboring RAM− neurons, whereas LY (300 nM) abolishes this difference. Frequency: control RAM+, 10.72 ± 0.86 Hz; control RAM−, 5.93 ± 1.13 Hz; LY (LY) RAM+, 4.88 ± 1.21 Hz; LY RAM−, 3.81 ± 0.5 Hz; F (1, 48) = 3.975, p = Amplitude: control RAM+, 13.47 ± 0.6 pA; control RAM−, 12.92 ± 0.92 pA; LY RAM+, 13.45 ± 1.05 pA; LY RAM−, 16.42 ± 1.3 pA; F (1, 48) = 2.829, p = 0.1. Control RAM+: n = 13, control RAM−: n = 11, LY RAM+: n = 13, LY RAM−: n = 15; 6 animals.
(D) 2D morphological analysis of RAM+ and RAM− neurons. (i) Schematic showing the sparse labeling strategy in CA3 pyramidal cells with RAM+ cells labeled in orange (GFP and tdTomato) and RAM− cells labeled in green (GFP only). (ii) Representative image of CA3 following the labeling scheme in (i). (iii) Representative images of dendritic segments with TEs of RAM+ and RAM− cells. (iv) 2D quantification of TE areas. TE area: RAM+, 1.52 ± 0.17, n = 12 dendrites, 5 animals; RAM−, 1.03 ± 0.11, n = 12 dendrites, 5 animals; p =
(E) Npas4 cKO mice were co-injected with AAV-RAM-mKate2 and AAV-hSyn-ΔCre-GFP. RAM+ CA3 pyramidal neurons have a significantly higher mEPSC frequency than their neighboring RAM− neurons, whereas LY (300 nM) abolishes this difference. Frequency: control RAM+, 6.31 ± 0.91 Hz; control RAM−, 2.88 ± 0.82 Hz; LY RAM+, 1.95 ± 0.54 Hz; LY RAM−, 2.19 ± 0.51; F (1, 23) = 5.653, p = Amplitude: control RAM+, 13.36 ± 0.67 pA; control RAM−, 14.2 ± 1.01 pA; LY RAM+, 15.21 ± 2.25 pA; LY RAM-, 14.82 ± 1.28 pA; F (1, 23) = , p = Control RAM+: n = 7, control RAM−: n = 8, LY RAM+: n = 5, LY RAM−: n = 7; 5 animals.
(F) Npas4 cKO mice were co-injected with AAV-RAM-mKate2 and AAV-hSyn-Cre-GFP. The mEPSC frequency for both RAM+ and RAM− was elevated by Npas4 deletion, but the difference between them was abolished. Frequency: control RAM+, 9.19 ± 2.08 Hz; control RAM−, 8.73 ± 1.03 Hz; LY RAM+, 1.24 ± 0.19 Hz; LY RAM−, 2.57 ± 0.65 Hz; F (1, 26) = , p = Amplitude: control RAM+, 13.31 ± 0.9 pA; control RAM−, 13.81 ± 0.52 pA; LY RAM+, 11.58 ± 0.94 pA; LY RAM−, 14.17 ± 1.1; F (1, 26) = 1.39, p = Control RAM+: n = 9, control RAM−: n = 8, LY RAM+: n = 6, LY RAM−: n = 7; 3 animals.
Summary data are shown as mean ± SEM. In (C), (E), and (F), ∗∗p < 0.01, two-way ANOVA, Bonferroni’s post hoc test. In (D), ∗p < 0.05, Mann-Whitney U test. See also Figure S4.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions

8. Figure 7
Npas4 Is Not Required for Activity-Dependent Homeostatic Shrinkage of TE Structures
(A) Schematic showing the sparse labeling strategy in CA3 pyramidal cells with morphology in green (GFP) and NaChBac or NaChBacmut cells labeled in red (tdTomato). CA3 pyramidal cells also expressed nuclearly localized Cre-GFP or ΔCre-GFP as a control.
(B) Representative image of CA3 cells following the labeling scheme in (A). Bottom: high-magnification image of a cell expressing cytosolic GFP to label morphology (arrowhead) and nuclear Cre-GFP (∗); neighboring cells expressed only nuclear Cre-GFP (arrow). Cells also expressed mCherry in the presence of NaChBac or NaChBachMut. Scale bars, = 100 and 10 μm, respectively.
(C) Representative traces from electrophysiological recordings of uninfected cells and cells expressing NaChBac or NaChBacMut 10 days after virus injection.
(D) Representative images of dendritic segments with TE structures. Scale bars, 5 μm.
(E) 2D quantification of TE areas. TE area: ΔCre + NaChBac, 1.01 ± 0.09 μm2/μm, n = 21 cells; ΔCre + NaChBacMut, 1.37 ± 0.08 μm2/μm, n = 22 cells; Cre + NaChBac, 1.12 ± 0.07 μm2/μm, n = 20 cells; Cre + NaChBacMut, 2.07 ± 0.14, n = 19 cells. 3 mice per condition, F (1,78) = 9.343, p = ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < Two-way ANOVA with Tukey’s post hoc multiple comparisons test. Summary data are shown as mean ± SEM.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions

9. Figure 8
Npas4 Mediates Activity-Dependent Modulation of TEs through Its Transcriptional Target Plk2
(A) Npas4 controls the activity-dependent expression of Plk2. Cultured hippocampal neurons from Npas4flx/flx mice were infected with AAV1-hSyn-GFP-Cre (Cre) or AAV1-hSyn-GFP (GFP) virus on day in vitro 9 (DIV9) and then stimulated with 10 μM kainic acid on DIV15 for 3, 6, and 9 hr before being lysed for RT-PCR analysis. 0 hr represents the unstimulated conditions. mRNA levels were normalized to Tuj1 expression levels. Data were derived from 4 independent experiments. ∗p < 0.05, Mann-Whitney U test.
(B and C) Selective expression of Plk2 in CA3 reduces TE size in Npas4 KO mice, whereas expression of the dominant-negative form of Plk2 (dnPlk2) fails to increase TE size further. AAV-hSyn-Plk2-FLAG or AAV-hSyn-dnPlk2-FLAG were stereotaxically injected into the hippocampus of Npas4 KO mice.
(B) Representative images showing Plk2 and dnPlk2 overexpression and the corresponding TEs in CA3 of Npas4 KO mice. Scale bars correspond to 200 μm on the top and 5 μm on the bottom.
(C) Summary bar graphs of TE size analysis in Npas4 KO mice injected with AAV-hSyn-Plk2-FLAG or AAV-hSyn-dnPlk2-FLAG. TE area: Plk2, 0.71 ± 0.05 μm2/μm; dnPlk2, 0.99 ± 0.09 μm2/μm; p = 0.026; n = 20 and 19 neurons, respectively, 4 animals for each condition. ∗p < 0.05, Mann-Whitney U test.
(D and E) The effect of Npas4 expression in CA3 of Npas4 KO, which previously reduced the TE size to the WT level, was blocked by dnPlk2. Npas4 KO mice were stereotaxically injected with AAV-hSyn-ΔNpas4-HA, AAV-hSyn-Npas4-HA, or AAV-hSyn-Npas4-HA and AAV-hSyn-dnPlk2-FLAG.
(D) Representative images showing Npas4 (red) and Plk2 (green) expression in CA3 of Npas4 KO mice co-injected with AAV-hSyn-Npas4-HA and AAV-hSyn-dnPlk2-FLAG. The scale bar corresponds to 300 μm. A representative image of TEs is shown on the right, and the scale bar there corresponds to 5 μm.
(E) Summary bar graph of TE size in Npas4 KO mice infected with AAV-hSyn-ΔNpas4-HA or AAV-hSyn-Npas4-HA or co-infected with AAV-hSyn-Npas4-HA and AAV-hSyn-dnPlk2-FLAG. TE area: ΔNpas4, 1.25 ± 0.12 μm2/μm; Npas4, 0.86 ± 0.06 μm2/μm; Npas4 + dnPlk2, 1.15 ± 0.10 μm2/μm; F (2, 42) = 4.396; p = 0.019; n = 15 neurons, 3 animals per condition. ∗p < 0.05, one-way ANOVA, Dunnett’s post hoc test. The data for Npas4 and ΔNpas4 conditions are re-plotted from Figure 5D.
(F and G) Resetting TE size in Npas4 KO mice rescues the contextual memory deficit.
(F) Npas4 KO mice were stereotaxically injected with AAV1-hSyn-dnPlk2 or AAV1-hSyn-Plk2 in CA3. Five days after surgery, mice underwent CFC, and their contextual memories were tested 24 hr later.
(G) Summary bar graph showing that Npas4 KO mice injected with Plk2 displayed similar freezing levels as their WT littermates, whereas those injected with dnPlk2 continued to display impaired contextual memory. Freezing level: WT, 45.83% ± 6.85%; KO + dnPlk2, 14.32% ± 5.44%; KO + Plk2, 37.71% ± 7.24%; F (2, 23) = 7.135; p = ∗p < 0.05, ∗∗p < 0.01, one-way ANOVA, Tukey’s post hoc test.
The blue and brown horizontal lines in (C) and (E) indicate the TE areas seen in Npas4 WT and KO mice, respectively (Figure 5C). All summary data are shown as mean ± SEM. See also Figures S5–S7.
Neuron  , e5DOI: ( /j.neuron )
Copyright © 2018 Elsevier Inc. Terms and Conditions