Developmental Inhibition of Gsk3 Rescues Behavioral and Neurophysiological Deficits in a Mouse Model of Schizophrenia Predisposition  Makoto Tamura, Jun Mukai, Joshua A. Gordon, Joseph A. Gogos  Neuron  Volume 89, Issue 5, Pages 1100-1109 (March 2016) DOI: 10.1016/j.neuron.2016.01.025 Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 1 Developmental Inhibition of Gsk3 Rescues the Spatial Working Memory Deficit in Df(16)A+/− Mice (A) Spatial delayed non-match-to-sample T maze task. Each trial of the task comprised a sample phase and choice phase separated by a 10-s delay. R, reward. (B) Acquisition (days to criterion) as a function of genotype and treatment is shown. Student’s t test; n = 11 (WT + vehicle), 12 (Df(16)A+/− + vehicle), 13 (Df(16)A+/− + SB), and 8 (WT + SB); error bars indicate ±SEM; ∗p < 0.05 and ∗∗p < 0.01 throughout. Neuron 2016 89, 1100-1109DOI: (10.1016/j.neuron.2016.01.025) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 2 Gsk3 Inhibition Rescues Deficits in Theta-Frequency Synchrony (A) Coherence spectra between vHPC and mPFC by group. Shaded area indicates ±SEM. (B) Theta- and gamma-range coherences between vHPC and mPFC by group are shown. Theta: p = 0.0037 (WT versus Df(16)A+/−) and p = 0.014 (Df(16)A+/− versus Df(16)A+/− + SB). Gamma: p = 0.024 (WT versus Df(16)A+/−) and p = 0.10 (Df(16)A+/− versus Df(16)A+/− + SB). Student’s t test; n = 10 (WT + vehicle), 10 (Df(16)A+/− + vehicle), 12 (Df(16)A+/− + SB), and 6 (WT + SB). (C) Distributions of vHPC theta phases at which action potentials were observed for representative mPFC neurons of each genotype. PPC values are 0.0083 (WT), 0.00007 (Df(16)A+/−), and 0.0045 (Df(16)A+/− + SB). (D) Strength of phase-locking of mPFC neurons to theta oscillations for all recorded neurons is shown. p = 0.036 (WT versus Df(16)A+/−) and p = 0.013 (Df(16)A+/− versus Df(16)A+/− + SB), Student’s t test; n = 135 (WT), 89 (Df(16)A+/−), 163 (Df(16)A+/− + SB), and 162 (WT + SB) units. (E) Distributions of vHPC gamma phases at which action potentials were observed for representative mPFC neurons of each genotype. PPC values are 0.00041 (WT), −0.00026 (Df(16)A+/−), and 0.000098 (Df(16)A+/− + SB). (F) Strength of phase-locking of mPFC neurons to gamma oscillations for all recorded neurons is shown. p = 0.027 (WT versus Df(16)A+/−) and p = 0.063 (Df(16)A+/− versus Df(16)A+/− + SB), Student’s t test. Sample sizes are as in (D). See also Figures S1, S2, and S3. Neuron 2016 89, 1100-1109DOI: (10.1016/j.neuron.2016.01.025) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 3 Gsk3 Inhibition Restores Goal Arm Encoding in Df(16)A+/− Mice (A) Spatial maps of firing rate for example mPFC single units are shown. (B) Normalized firing rate (Z score, top right) and a raster plot (bottom right) of spikes fired by an example left arm-selective single unit across trials aligned to goal arm entrance are shown. (C) (Left) Percentage of units that were goal selective as a function of time from entering goal arm, according to rmANOVAs performed on binned spike rates. Percentages were calculated within animals, then averaged across animals. (Right) Mean percentage of units at time zero is shown. p = 0.016 (WT versus Df(16)A+/−) and p = 0.031 (Df(16)A+/− versus Df(16)A+/− + SB), Student’s t test. Dashed line represents chance (p = 0.05). (D) Firing rates by group. Firing rate did not differ by genotype (F1,1341 = 0.01, p = 0.94) or treatment (F1,1341 = 3.29, p = 0.070, two-way ANOVA). n = 412 units from 11 animals (WT), 297 units from 12 animals (Df(16)A+/−), 405 units from 13 animals (Df(16)A+/− + SB), and 231 from 8 animals (WT + SB). See also Figure S4. Neuron 2016 89, 1100-1109DOI: (10.1016/j.neuron.2016.01.025) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 4 Disrupted Population Coding in Df(16)A+/− Mice Is Reversed by Developmental Inhibition of Gsk3 (A) Schematic of run sequence. Spike data from all isolated mPFC units were structured as peri-event spike histograms centered on key events during the task as follows: leaving start box (a), entering goal arm (b), reaching goal port (c), leaving goal arm (d), and returning to start box (e). (B) Accuracy of choice goal decoding during choice run in an individual WT mouse. Solid lines denote mean decoding accuracy, and shaded areas cover 95% confidence intervals for real (red) and shuffled (gray) data. (C) Decoding accuracy of choice goal as a function of time from entering goal arm, computed for individuals and averaged across animals by group. Solid lines denote mean decoding accuracy across animals, and shaded areas cover SEM. n = 10 (WT), 7 (Df(16)A+/−), 9 (Df(16)A+/− + SB), and 8 (WT + SB). Dashed line represents chance level. (D) Mean decoding accuracy of goal representations at time zero is shown. p = 0.0092 (WT versus Df(16)A+/−) and p = 0.029 (Df(16)A+/− versus Df(16)A+/− + SB), Student’s t test. Sample sizes are as in (C). See also Figure S4. Neuron 2016 89, 1100-1109DOI: (10.1016/j.neuron.2016.01.025) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 5 Spatial Information during Retrieval Correlates with vHPC-mPFC Synchrony during Encoding (A) Raster plot across trials (bottom) and MI for goal location computed on binned spike rates (top) for an example unit, aligned to goal arm entry during the choice phase, are shown. (B) Mean MI at goal arm entry for all neurons, by genotype and treatment, is shown. p = 0.0041 (WT versus Df(16)A+/−) and p = 0.00023 (Df(16)A+/− versus Df(16)A+/− + SB), Student’s t test. n = 412 units from 11 animals (WT), 297 units from 12 animals (Df(16)A+/−), and 405 units from 13 animals (Df(16)A+/− + SB). (C) Scatterplots depict MI during the choice phase, plotted against the strength of phase-locking of mPFC units to vHPC gamma during the sample phase, for all neurons recorded from a representative WT (left) and Df(16)A+/− (right) mouse. (D) Correlation between MI measured during the sample phase and phase-locking measured during the sample (left) and choice (right) phases, averaged across animals, is shown (differences from r = 0 by t test: WT, n = 10 mice, p = 0.029; Df(16)A+/−, n = 7, p = 0.65; and Df(16)A+/− + SB, n = 9, p = 0.15 for sample; WT, p = 0.37; Df(16)A+/−, p = 0.81; and Df(16)A+/− + SB, p = 0.27 for choice). (E) Normalized MI as a function of phase-locking strength to vHPC gamma, binned by quartiles, is shown (multiple linear regression, p = 0.028 (WT), 0.32 (Df(16)A+/−), and 0.11 (Df(16)A+/− + SB). (F) MI as a function of phase-locking strength, binned into quartiles and normalized by the mean of the first quartile, is shown (paired t test, p = 0.036 (WT), p = 0.84 (Df(16)A+/−), and p = 0.024 (Df(16)A+/− with SB). See also Figure S5. Neuron 2016 89, 1100-1109DOI: (10.1016/j.neuron.2016.01.025) Copyright © 2016 Elsevier Inc. Terms and Conditions