Alterations in DNA methylation rates of brain-derived neurotrophic factor in patients with schizophrenia.

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Figure captions

Fig.1. Schema of CpGs and primers used for DNA methylation analyses.

The target region used for methylation analysis consists of 1092 bp including 81 CpGs

upstream of exon I.

Fig.2. Hierarchic cluster analysis of subjects and their methylation profiles at CpG

I of the BDNF gene.

Two-way hierarchic cluster analysis of 44 samples (rows) and DNA methylation of CpG

units at CpG I of the BDNF gene (columns) are shown. DNA methylation values are

depicted by a pseudocolor scale as methylation rate increases from red (nonmethylated)

to yellow (methylated). White denotes data of poor quality, and such data points were

excluded before clustering. Samples are color-coded according to the diagnoses of

samples (depicted in legend at upper left).

Table 1. Demographic and clinical characteristics of subjects.

Group

Age (Years)

(Mean ±

S.D.)

SZ (N=22)

36.4 ± 11.8

Control (N=22)

41.1 ± 13.4

Duration of

illness

(Years)

(Mean±S.D.)

14.7 ± 13.0

Dosage of daily

antipsychotics (mg)

(mean ± S.D.)

BPRS

(mean ±

S.D.)

753 ± 851

61.5 ± 16.2

All participants were male Japanese.

Dosage of daily antipsychotics was represented by chlorpromazine equivalent dose.

BRPS: Brief Psychiatric Rating Scale score.

Table 2. The DMRs of each CpG units at the CpG island of BNDF exon I in

schizophrenic patients and control.

Control

SZ

ANCOVA

Rate (%)

(Mean ± S.E.M.)

Rate (%)

(Mean ± S.E.M.)

p-value

CpG_1,7

9.4 ± 0.6

5.1 ± 0.6

<0.0001*

CpG_2

CpG_3,6

5.6 ± 0.5

8.1 ± 0.5

3.5 ± 0.7

7.5 ± 0.9

0.021

0.54

CpG_4

CpG_5

23.3 ± 3.2

2.3 ± 0.3

31.7 ± 3.2

1.8 ± 0.4

0.064

0.32

CpG_8,9

10.2 ± 0.4

13.2 ± 0.5

<0.0001*

CpG_14

7.3 ± 0.6

12.6 ± 1.8

0.010

CpG_15

9.8 ± 0.4

7.0 ± 0.8

0.0051

CpG_17

CpG_18

6.4 ± 0.5

2.5 ± 0.4

3.3 ± 0.7

0.8 ± 0.2

0.0008*

0.0004*

CpG_19,20,21

5.1 ± 0.5

3.3 ± 0.4

0.0054

CpG_22

13.5 ± 1.2

22.2 ± 3.9

0.038

CpG_23

CpG_24

3.8 ± 0.4

1.4 ± 0.1

3.4 ± 0.6

1.4 ± 0.2

0.57

0.82

CpG_25,26,27

3.7 ± 0.2

3.3 ± 0.2

0.23

CpG_28

3.0 ± 0.2

3.0 ± 0.3

0.84

CpG_29,30,31

2.2 ± 0.1

1.8 ± 0.2

0.034

CpG_32

CpG_33,34

11.7 ± 1.4

7.8 ± 0.4

13.5 ± 0.9

10.2 ± 1.0

0.27

0.030

CpG_36

CpG_37

5.6 ± 1.0

4.7 ± 0.3

4.3 ± 0.9

5.4 ± 0.4

0.36

0.25

CpG_47

CpG_48

3.0 ± 0.3

3.7 ± 0.2

3.1 ± 0.3

4.9 ± 0.6

0.80

0.055

CpG_50,51

CpG_52

3.7 ± 0.3

3.0 ± 0.4

4.7 ± 0.3

2.4 ± 0.3

0.018

0.20

CpG_59

7.3 ± 1.5

5.4 ± 0.5

0.26

CpG_61

CpG_63,77

4.8 ± 0.4

2.2 ± 0.4

4.6 ± 0.6

1.8 ± 0.4

0.75

0.44

CpG_71

CpG_72,73

21.9 ± 3.6

4.1 ± 0.4

28.9 ± 4.7

3.9 ± 0.4

0.28

0.77

CpG_74,75

3.8 ± 0.4

2.6 ± 0.2

0.011

CpG_78

16.7 ± 2.5

11.6 ± 1.8

0.11

CpG_80,81

6.0 ± 0.4

6.0 ± 0.5

0.98

The asterisks (*) behind of scores indicate statistically significant P-values.

Significance was set at P<0.0015

Table 3. Correlations of the DMRs at the CpG island of BNDF exon I with a

between the DMRs and dosage of daily antipsychotics in patients with SZ.

Correlation coefficient

p-value

CpG_1,7

0.28

0.21

CpG_2

CpG_3,6

0.21

-0.048

0.35

0.83

CpG_4

-0.41

0.061

CpG_5

CpG_8,9

0.31

-0.079

0.16

0.73

CpG_14

CpG_15

-0.009

0.002

0.97

0.99

CpG_17

CpG_18

-0.46

0.19

0.031

0.39

CpG_19,20,21

CpG_22

0.16

-0.21

0.47

0.36

CpG_23

-0.33

0.14

CpG_24

CpG_25,26,27

-0.32

-0.11

0.14

0.63

CpG_28

CpG_29,30,31

-0.71

-0.30

0.00022 **

0.17

CpG_32

0.24

0.28

CpG_33,34

-0.025

0.91

CpG_36

CpG_37

0.11

-0.40

0.63

0.068

CpG_47

-0.45

0.037

CpG_48

-0.081

0.72

CpG_50,51

0.12

0.59

CpG_52

CpG_59

-0.38

-0.31

0.079

0.17

CpG_61

CpG_63,77

-0.13

-0.052

0.56

0.82

CpG_71

CpG_72,73

0.009

-0.19

0.97

0.40

CpG_74,75

-0.21

0.34

CpG_78

0.057

0.80

CpG_80,81

-0.52

0.013

Correlation coefficient and p-value by semi-partial correlation coefficients

procedure is shown.

The asterisks (*) behind of scores indicate statistically significant p-values.

Significance was set at p<0.0015.

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