Journal of Pharmacology and Experimental Therapeutics

JPET#178475

The efficacy of sodium channel blockers to prevent PCP-induced cognitive dysfunction in the rat: Potential for novel treatments for schizophrenia

Charles H. Large, Silvia Bison, Ilaria Sartori, Kevin D. Read, Alessandro Gozzi, Davide Quarta, Marinella Antolini, Emma Hollands, Catherine H. Gill, Martin J. Gunthorpe, Nagi Idris, Jo C. Neill, Giuseppe S. Alvaro.

Supplementary Figure 1: The influence of GSK2 (20-80mg/kg p.o.) on performance of the initial and reversal phase of the reversal learning task. Data are expressed as mean ± S.E.M. of % correct responding (n=10 per group).

100 Initial phase Reversal phase 90 80 g n 70i d n

60o p s

40c e r r

30o c 20 % 10 0 VEH 20 40 60 80 VEH 20 40 60 80

Supplementary Table 1: The effect of GSK2 (20-80mg/kg) on the total number of lever presses in a reversal learning paradigm. Data are expressed as mean  S.E.M. of total number of lever presses (n=10 per group) in the initial and reversal phase.

Drug treatment Initial phase Reversal phase vehicle 26.6± 0.3 26.4± 0.3 GSK2 (20 mg/kg) 27.7± 0.3 27.8± 0.2 GSK2 (40 mg/kg) 27.7± 0.2 27.5± 0.2 GSK2 (60 mg/kg) 27.5± 0.3 27.3± 0.2 GSK2 (80 mg/kg) 26.2± 0.1 26.2± 0.1 JPET#178475

Supplementary Table 2: The effect of GSK2 (40-80mg/kg, p.o.) and PCP (1.5mg/kg, i.p.) on the total number of lever presses in the reversal learning paradigm. Data are expressed as the mean ± S.E.M. of total number of lever presses (n=9-10) per group) in the initial and reversal phase of the task.

Drug treatment Initial phase Reversal phase vehicle + vehicle 26.7 ± 0.2 26.5 ± 0.3 vehicle + PCP (1.5mg/kg) 26.9± 0.2 26.9± 0.2 GSK2 (40mg/kg) + PCP 26.8± 0.3 26.6± 0.2 GSK2 (60mg/kg) + PCP 26.6± 0.3 26.0± 0.5 GSK2 (80mg/kg) + PCP 26.9± 0.4 26.3± 0.2

Supplementary Figure 2: The influence of GSK13 (10-60mg/kg, p.o.) on performance of the initial and reversal phase in a reversal learning task. Data are expressed as mean ± S.E.M. of % correct responding (n=10 per group).

100 Initial phase Reversal phase 90

g 80 n i

d 70 n o

p 60 s e

e 40 r r

o 30 c 20 % 10 0 veh 10 30 60 veh 10 30 60

Supplementary Table 3: The effect of GSK1061436A (10-60mg/kg, p.o.) on the total number of lever presses in a reversal learning paradigm. Data are expressed as mean ± S.E.M. of total number of lever presses (n=10 per group) in the initial and reversal phase.

Drug treatment Initial phase Reversal phase vehicle 27.4± 0.3 27.4± 0.2 GSK3 (10mg/kg) 26.2± 0.4 26.1± 0.6 GSK3 (30mg/kg) 26.1± 0.3 26.8± 0.2 GSK3 (60mg/kg) 26.2± 0.4 26.8± 0.3 JPET#178475

Supplementary Table 4: The effect of GSK3 (10-60mg/kg, p.o.) and acute PCP (1.5mg/kg, i.p.) on the total number of lever presses in a reversal learning paradigm. Data are expressed as the mean ± S.E.M. of total number of lever presses (n=10) per group in the initial and reversal phase of the task.

Drug treatment Initial phase Reversal phase vehicle + vehicle 26.4± 0.3 26.5± 0.2 vehicle + PCP (1.5mg/kg) 26.1± 0.3 26.8± 0.5 GSK3 (10mg/kg) + PCP 26.2± 0.3 26.1± 0.5 GSK3 (30mg/kg) + PCP 27.1± 0.3 26.3± 0.6 GSK3 (60mg/kg) + PCP 25.8± 0.4 27.7± 0.2

Supplementary Table 5: The effect of lamotrigine (25mg/kg, i.p.) and acute PCP (1.5mg/kg, i.p.) on the total number of lever presses in a reversal learning paradigm. Data are expressed as the mean ± S.E.M. of total number of lever presses (n=10) in the initial and reversal phase of the task.

Drug treatment Initial phase Reversal phase vehicle + vehicle 26.2± 0.3 27.1± 0.2 vehicle + PCP 1.5mg/kg 26.4± 0.3 26.8± 0.2 lamotrigine (25mg/kg) + vehicle 26.5± 0.2 26.5± 0.2 lamotrigine (25mg/kg) + PCP 27.5± 0.2 27.1± 0.2 JPET#178475

Supplementary Table 6: Mean group weights and arterial blood gases measurements. paCO2 - partial pressure of arterial CO2; PaO2 - partial pressure of arterial O2; values “pre” and “post” were recorded prior to and at the end of phMRI timeseries, respectively. Values expressed as mmHg and presented as mean ± S.E.M.. Pharmacological treatments received by each group are described in the methods.

Group Weight(g) paCO2 pre paCO2 post paO2 pre paO2 post

#1 (N=9) 274±7 38±3 41±2 153±10 146±5 #2 (N=7) 292±11 38±1 37±2 137±11 140±6 #3 (N=6) 297±5 41±2 42±3 118±10 139±9 #4 (N=6) 283±10 42±2 37±2 130±14 148±7 #5 (N=5) 314±12 40±2 35±2 191±8 186±5 #6 (N=7) 282±11 38±2 36±2 176±7 182±4 #7 (N=7) 279±13 39±2 40±2 194±8 178±5 #8 (N=6) 306±12 40±2 36±2 199±8 207±5 JPET#178475

Supplementary Figure 3: rCBV timecourse following PCP injection in representative VOIS. PCP was administered at time 0, corresponding to 30 minutes after s.c. pretreatment with vehicle (group 5) or GSK2 (group 6 and 7). Baseline data were obtained in animals pretreated and challenged with saline (group 8). Data are plotted as mean ± S.E.M. within each group. [Cg: cingulate cortex; VHc: ventral hippocampus; DLTh: dorsolateral thalamus, Ss: somatosensory cortex].

Supplementary Figure 4: rCBV timecourse following PCP injection in representative VOIS. PCP was administered at time 0, corresponding to 30 minutes after i.p. pretreatment with vehicle (group 1) or GSK3 (groups 2 and 3). Baseline data were obtained in animals pretreated and challenged with saline (group 8). Data are plotted as mean ± S.E.M. within each group. [Cg: cingulate cortex; VHc: ventral hippocampus; DLTh: dorsolateral thalamus, Ss: somatosensory cortex].

JPET#178475

Supplementary Figure 5: rCBV timecourse following PCP injection in representative VOIS. PCP was administered at time 0, corresponding to 30 minutes after i.p. pretreatment with vehicle (group 1) or lamotrigine (group 4). Baseline data were obtained in animals pretreated and challenged with saline (group 8). Data are plotted as mean ± S.E.M. within each group. [Cg: cingulate cortex; VHc: ventral hippocampus; DLTh: dorsolateral thalamus, Ss: somatosensory cortex]. JPET#178475

Supplementary Figure 6: Effect of pretreatment with GSK1014802 (10 or 30 mg/kg, groups 6 and 7) or vehicle (group 5) on rCBV baseline in four representative brain regions. GSK1014802 or vehicle M100907 were administered at time 0. Data are plotted as mean ± S.E.M. within each group. [Cg: cingulate cortex; VHc: ventral hippocampus; DLTh: dorsolateral thalamus, Ss: somatosensory cortex]. JPET#178475

Supplementary Figure 7: Effect of pretreatment with GSK3 (groups 2 and 3) or vehicle (group 1) on rCBV baseline in four representative brain regions. GSK3 or vehicle (saline) were administered SC at time 0. Data are plotted as mean ± S.E.M. within each group. [Cg: cingulate cortex; VHc: ventral hippocampus; DLTh: dorsolateral thalamus, Ss: somatosensory cortex]. JPET#178475

Supplementary Figure 8: Effect of pretreatment with lamotrigine (group 4) or vehicle (group 1) on rCBV baseline in four representative brain regions. Lamotrigine or vehicle were administered IP at time 0. Data are plotted as mean ± S.E.M. within each group. [Cg: cingulate cortex; VHc: ventral hippocampus; DLTh: dorsolateral thalamus, Ss: somatosensory cortex]. JPET#178475

Supplementary Figure 9: The effect of (a) GSK2 (100 M) and (b) GSK3 (100 M) on whole- cell patch-clamp recordings of NMDA receptor-mediated responses, recorded from rat dissociated cortical neurons.