SUPPLEMENTARY INFORMATION

Supplementary Materials and Methods

Animals. We used Shn2 KO mice (Takao et al, 2013), Cn KO mice (Cottrell et al, 2013; Miyakawa et

al, 2003; Suh et al, 2013; Zeng et al, 2001), Nrgn KO mice (Huang et al, 2006; Huang and Huang,

2012; Pak et al, 2000), Camk2a HKO mice (Hagihara et al, 2016; Yamasaki et al, 2008), Chd8 HKO

mice (Katayama et al, 2016), Disc1-L100P mutant, Disc1-Q31L mutant mice (Shoji et al, 2012), Barp

KO mice (Nakao et al, 2015), and their corresponding control mice. Table S2 summarizes the

behavioral phenotypes of these mutant mice.

Shn2 KO mice. Shn2 was originally identified as a nuclear factor-κB (NF-κB) site-binding protein that

tightly binds to the enhancers of major histocompatibility complex (MHC) class I genes and acts as

an endogenous inhibitor of NF-κB (Fukuda et al, 2002). Deficiencies in Shn2 may cause mild

chronic inflammation in the brain and confer molecular, neuronal, and behavioral phenotypes

relevant to schizophrenia in mice (Takao et al, 2013). Genome-wide association studies (GWASs)

have identified a number of single nucleotide polymorphisms (SNPs) in the MHC region associated

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with schizophrenia (Purcell et al, 2009; Shi et al, 2011; Stefansson et al, 2009). Shn2 KO mice

exhibit multiple abnormal behaviors related to schizophrenia, including increased locomotor

activity, deficits in working memory, abnormal social behavior, and impaired prepulse inhibition,

which are commonly observed in Cn KO mice (Miyakawa et al, 2003; Zeng et al, 2001) and Nrgn KO

mice (Hattori et al, 2015; Huang et al, 2006; Huang and Huang, 2012; Miyakawa et al, 2001; Pak et

al, 2000) as well (Table S2).

Cn KO mice. Calcineurin (Cn) is a calcium-dependent protein phosphatase and has been implicated

in synaptic plasticity (Winder and Sweatt, 2001). CN has been reported to be associated with

schizophrenia (Gerber et al, 2003; Liu et al, 2007; Sacchetti et al, 2013), and altered expression of

calcineurin has been observed in the postmortem brains of patients with schizophrenia (Eastwood

et al, 2005; Wada et al, 2012). Forebrain-specific Cn KO mice exhibit behavioral and cognitive

abnormalities related to schizophrenia, including increased locomotor activity, deficits in working

memory, abnormal social behavior, and impaired prepulse inhibition (Miyakawa et al, 2003; Zeng

et al, 2001; unpublished data) (Table S2). Deficits in synaptic transmission in the frontal cortex have

been suggested to be the underlying mechanism of working memory impairment in these mice

(Cottrell et al, 2013). In addition, Cn KO mice exhibit disruption in ripple-associated information

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processing in the hippocampal CA1, which is implicated in cognitive impairments associated with

schizophrenia (Suh et al, 2013).

Nrgn KO mice. Neurogranin (Nrgn) is a calmodulin-binding protein that modulates activity of the

Camk2 protein downstream of N-methyl-d-aspartic acid (NMDA) receptors, and is implicated in

synaptic plasticity (Pak et al, 2000). GWAS revealed significant association with SNPs located

upstream of NRGN (Stefansson et al, 2009), a finding recently confirmed by a large-scale GWAS

(Schizophrenia Working Group of the Psychiatric Genomics Consortium, 2014), strongly suggesting

that NRGN is a susceptibility gene for schizophrenia. Nrgn KO mice exhibit behavioral phenotypes

related to schizophrenia, including increased locomotor activity, deficits in working memory,

abnormal social behavior, and impaired prepulse inhibition (Hattori et al, 2015; Huang et al, 2006;

Huang and Huang, 2012; Miyakawa et al, 2001; Pak et al, 2000) (Table S2).

Camk2a HKO mice. Camk2 is a major downstream molecule of the NMDA receptor and is thought

to play an essential role in synaptic plasticity. A recent study demonstrated genetic association of

CAM2KA with bipolar disorder (Ament et al, 2015), and decreased mRNA expression has been

observed in the frontal cortex of patients with bipolar disorder (Xing et al, 2002). In addition, the

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Camk2a gene was identified as one of the top candidate genes for bipolar disorder by a meta-

analysis that integrated genetic and genomic data from both human and animal studies (Le-

Niculescu et al, 2009). At the cellular level, neuronal hyperexcitability, which we previously

detected in the hippocampal granule cells of Camk2a HKO mice (Yamasaki et al, 2008), was also

found in the granule cell-like neurons differentiated from induced pluripotent stem cells (iPSCs)

derived from patients with bipolar disorder (Mertens et al, 2015). Camk2a HKO mice exhibit

abnormal behaviors, such as hyper-locomotion, deficits in social activity and working memory,

which are analogous to those in patients with bipolar disorder/schizophrenia (Shin et al, 2013;

Yamasaki et al, 2008) (Table S2). In addition, these mutant mice exhibit periodic changes in

locomotor activity in their home cages with an approximate cycle length of 10–20 days. These

changes in locomotor activity are associated with fluctuations of anxiety-like and depression-like

behaviors, suggesting that the mutant mice exhibit infradian oscillations of mood substantially

similar to those observed in patients with bipolar disorder (Hagihara et al, 2016). Carbamazepine,

a mood stabilizer, partially normalized this exaggerated infradian rhythm of the mice. However,

lithium, the most effective mood stabilizer available, failed to treat the phenotype. Obvious

depressive episode-like behavioral changes have not been observed to date in these mutant mice.

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Therefore, Camk2a HKO mice could recapitulate some aspects of bipolar disorder and may serve as

a unique animal model of recurrent manic episode of bipolar disorder.

Chd8 HKO mice. Chd8, a member of the chromodomain helicase DNA-binding family of proteins, is

known to act as a chromatin-remodeling factor. Recent exome sequencing analyses have identified

a number of de novo mutations in a variety of genes in individuals with ASD, further revealing that

CHD8 is the most frequently affected gene (Neale et al, 2012; O’Roak et al, 2012a, 2012b;

Talkowski et al, 2012). CHD8 has been implicated also in schizophrenia (Kimura et al, 2016;

McCarthy et al, 2014). Chd8 HKO mice exhibit behavioral abnormalities reminiscent of ASD in

humans, including increased anxiety, increased persistence, and abnormal social interaction

(Katayama et al, 2016) (Table S2). Chd8 deficiency induces aberrant activation of RE1 silencing

transcription factor (REST), a molecular brake of neuronal development, resulting in

neurodevelopment abnormalities in mice (Katayama et al, 2016).

Disc1-L100P and Disc1-Q31L mutant mice. DISC1 has been regarded as a putative susceptibility

gene for psychiatric disorders such as schizophrenia, bipolar disorder, and major depressive

disorder (Craddock et al, 2006). However, a recent large-scale analysis of copy number variants

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suggested that DISC1 may not be a risk factor for the disorder (Marshall et al, 2016). Previous

studies have reported that N–ethyl–N–nitrosourea (ENU)-mutagenized Disc1-L100P and Disc1-

Q31L mutant mice exhibit schizophrenia-like and depression-like behaviors, respectively (Clapcote

et al, 2007); however, some research groups failed to observe such behavioral abnormalities in

these mutant mice (Arime et al, 2014; O’Tuathaigh et al, 2017; Shoji et al, 2012) (Table S2). This

discrepancy may be explained by differences in the genetic background, breeding and rearing

conditions, test protocol, and/or age and time at the testing (Shoji et al, 2012). In the present

study, we used brains of Disc1-L100P and Disc1-Q31L mutant mice, which did not exhibit

behavioral abnormalities related to psychiatric disorders (Table S2).

Barp KO mice. Genes encoding voltage-gated Ca2+ channels (VGCCs) (e.g., CACNA1C, CACNB2, and

CACNA1l) have been implicated in the pathogenesis of schizophrenia and other psychiatric

disorders (Schizophrenia Working Group of the Psychiatric Genomics Consortium, 2014).

Deficiency of Barp, a VGCC activity-regulating molecule, induces multiple behavioral phenotypes

that are seemingly opposite to those observed in the mouse models of schizophrenia and related

disorders. Such changes include increased working memory, prepulse inhibition, and social

interaction as well as decreased locomotor activity, although statistical findings for many of these

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phenotypes are weak (Nakao et al, 2015) (Table S2).

Collectively, these findings indicate that Shn2 KO, Cn KO, Nrgn KO, Camk2a HKO, and Chd8 HKO

mice—but not Disc1-L100P and Disc1-Q31L mutant, or Barp KO mice—exhibit good construct and

face validities for their respective disorders.

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Table S1. Patient characteristics. Antipsychotic dose (mg) is measured as chlorpromazine

equivalents in the dataset of Dean et al. and as fluphenazine equivalents in the other datasets. M,

male; F, female; na, not available.

Table S2. Behavioral characteristics of the mutant mice used in this study. Arrays represent an

increase or decrease in a comparison between the mutant and control mice.

Table S3. Genes whose expression was altered in the brains of mouse models of psychiatric

disorders. Genes whose expression was altered in at least four out of eight mouse datasets were

processed for pathway analyses.

Table S4. Pathway analyses of the genes whose expression was altered in the brains of mouse

models of the psychiatric disorders using DAVID, ADGO, and GoToolBox. The top 20 pathways

(ranked based on the P-value) are shown for each analysis.

Table S5. Expression patterns of genes encoding enzymes related to glycolysis pathway in the

brains of mouse models of psychiatric disorders.

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Figure S1. Lower pH in the postmortem brains of patients with schizophrenia and bipolar

disorder revealed by a Z-score-based meta-analysis. Bar graph of the Z-score transformed pH

values (average ± SEM). Results of a Tukey’s HSD post hoc test followed by a two-way ANOVA are

shown in this figure. *1P = 0.020, *2P < 0.0001, *3P = 0.0001, *4P = 0.027; ANOVA/Tukey’s post hoc

test within each dataset.

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Figure S2. Correlation analyses between brain pH and postmortem interval, and age. Scatter

plots of pH versus postmortem interval or age in the schizophrenia datasets (a, b) and bipolar

disorder datasets (c, d). The Pearson’s r and P values are shown for each panel.

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Figure S3. No significant correlations between brain pH and lifetime antipsychotic use in patients

with schizophrenia and bipolar disorder revealed by Z-score-based meta-analysis. Scatter plot of

Z-score-transformed values of pH versus those of lifetime antipsychotic use in the six datasets.

SMRI: Stanley Medical Research Institute.

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Figure S4. Correlation analyses between age and pH/lactate levels in the brains of mouse models

of psychiatric disorders. Scatter plots of pH (row values) versus age in control (a) and mutant (b)

groups. Scatter plots of lactate levels (row values) versus age in control (c) and mutant (d) groups.

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Figure S5. No significant differences in pH and lactate levels were observed in mutant mice that

did not exhibit behavioral abnormalities associated with psychiatric disorders. Bar graphs of pH

(a) and lactate levels (b) in the brains of Disc1-L100P and Disc1-Q31L mutant mice, Barp KO mice,

and their corresponding controls (mean ± SEM). Each plot represents individual mouse values.

Unadjusted P-values (Student’s t-test) are shown.

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Figure S6. Potentially elevated glycolysis in the brains of mouse models of psychiatric disorders.

Glycolysis-related genes whose expression was altered in the brains of mouse models of psychiatric

disorders were mapped in a schematic of the glycolysis pathway

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