epidemiological, behavioral and MRI data

epidemiological, behavioral and MRI data by each subjec

Results of conjunctional VBM analysis of T contrasts.

<p>All statistical parametric maps were thresholded at P<0.001 with a cluster level K>20, age was added as a covariate. In column diagrams, beta-values are given for each cluster. Significant contrasts (P<0.05) are marked with star (*). <i>Left:</i> contrast TD>ASD in a coronal view (Y = −5); <i>Middle:</i> contrast TD>ASD in a saggital view (X = −2); <i>Right:</i> contrast SCZ>ASD in a coronal view (Y = 14).</p

Summary of main effect results and between group analyses in MNI space on a p<0.001 level.

<p>Summary of main effect results and between group analyses in MNI space on a p<0.001 level.</p

Correlation plots.

<p>Left: Correlation between left amygdala GM volume (beta-values) and the ‘reading the mind in the eyes’-score (RME) in autism spectrum disorder (r = 0.41; P = 0.015). The RME-score represents the number of correctly identified emotions of pictures depicting facial expressions (maximum score = 28). Right: Correlation between left insular GM volume (beta-values) and the PANSS hallucinatory behaviour score in schizophrenia (r = 0.56; P = 0.008). The hallucinatory behaviour score represents the degree of a patient’s verbal report or behaviour indicating perceptions which are not generated by external stimuli. The score ranges between 1 (absent) and 7 (extreme).</p

Sociodemographical and clinical parameters of ASD, SCZ and TD.

<p>Significance threshold was defined as P<0.05.</p>a<p>PANSS = Positive and Negative Syndrome Scale; p = positive scale; ns = negative scale; gps = general psychopathology scale; t = total;</p>b<p>ADOS = Autism Diagnostic Observery Scale; missing data in 2 cases; c = communication; si = social interaction; t = total;</p>c<p>ADI-R = Autism Diagnostic Interview - Revised; missing data in 2 cases; si = social interaction; cl = communication and language; rb = restricted and repetitive behaviours;</p>d<p>RME = ’Reading the Mind in the Eyes’ test,</p>e<p>CPZ-eq = Chlorpromazine equivalents.</p><p>Sociodemographical and clinical parameters of ASD, SCZ and TD.</p

Box plots of global brain measures.

<p>Box plots are given for autism spectrum disorder (ASD), schizophrenia (SCZ) and typically developed control subjects (TD) each with global gray matter (GM), global white matter (WM) and total brain volume (TBV). The horizontal line in each box indicates the median, while the top and bottom borders of the box mark the 25th and 75th percentile, respectively. The vertical lines above and below the box mark the range of distribution.</p

Genetic alterations identified in the control subject SWE_Q56_508.

<p>A. <i>SHANK2</i> splice mutation (IVS22+1G>T) detected in a Swedish female control, SWE_Q56_508. The mutation altered the donor splicing site of exon 22 and led to a premature stop in all <i>SHANK2</i> isoforms except for the <i>AF1411901</i> isoform, where it altered the protein sequence (G263V). B. CNVs in the same individual altering <i>LOC339822</i>, <i>SNTG2</i>, <i>PXDN</i> and <i>MYT1L</i>. The two close duplications span 264 kb and 245 kb on chromosome 2 and altered <i>LOC339822</i> and <i>SNTG2</i>, and <i>PXDN</i> and <i>MYT1L</i>, respectively. Dots show the B allele frequency (BAF; in green), Log R ratio (LRR; in red), and QuantiSNP score (in blue). Lower panel: all CNVs listed in the Database of Genomic Variants (DGV) are represented: loss (in red), gain (in blue), gain or loss (in brown). H, homer binding site; D, dynamin binding site; C, cortactin binding site.</p

Characterization of the functional impact of <i>SHANK2</i> mutations in cultured neuronal cells.

<p>A. The colocalization of <i>ProSAP1A/SHANK2</i>-EGFP (postsynaptic marker) and Bassoon (presynaptic marker) indicated that the mutations did not disturb the formation of SHANK2 clusters at excitatory synapses along the dendrites. B. The quantification of synapse density was performed on 20 transfected hippocampal neurons per construct from at least three independent experiments. The majority of the <i>ProSAP1A</i> variants affecting a conserved amino acid among SHANK proteins reduced significantly the synaptic density compared with the variants that affect amino acid non conserved among SHANK proteins (Mann-Whitney U-test: n_WT = 20, n_mut = 20; U_S557N = 82.5, p_S557N = 0.001; U_R569H = 124, p_R569H = 0.04; U_L629P = 149, p_L629P = 0.17; U_V717F = 114, p_V717F = 0.02; U_A729T = 73, p_A729T = 0.000; U_K780Q = 154, p_K780Q = 0.221; U_R818H = 108, p_R818H = 0.012; U_A822T = 154.5, p_A822T = 0.224; U_V823M = 129, p_V823M = 0.056; U_Y967C = 134, p_Y967C = 0.076; U_G1170R = 78, p_G1170R = 0.001; U_R1290W = 142, p_R1290W = 0.121; U_Q1308R = 162, p_Q1308R = 0.314; U_D1535N = 97, p_D1535N = 0.005; U_P1586L = 137, p_P1586L = 0.910; U_L1722P = 79, p_L1722P = 0.001, *p<0.05, **p<0.01, ***p<0.001). <b>C.</b> Effect of the variants on synaptic density. The y-axis represents −log P compared to WT (P obtained with Mann-Whitney test). After Bonferroni correction for 16 tests, only P values<0.003 were considered as significant. Variants represented in red were specific to ASD, in orange were shared by ASD and controls, and in green were specific to the controls. Open circles and filled circles represent non conserved and conserved amino acids, respectively. Prim, primary; second, secondary.</p

Genomic structure, isoforms, and expression of human <i>SHANK2</i>.

<p>A. Genomic structure of the human <i>SHANK2</i> gene. Transcription of <i>SHANK2</i> produces four main mRNA from three distinct promoters: <i>SHANK2E</i> (<i>AB208025</i>), <i>ProSAP1A</i> (<i>AB208026</i>), <i>ProSAP1</i> (<i>AB208027</i>) and <i>AF141901</i>. There are three translation starts: in exon 2 for <i>SHANK2E</i>, in exon1b for <i>ProSAP1A</i>, and in exon1c for <i>ProSAP1</i> and <i>AF141901</i>; and two independent stop codons: in exon 22b for <i>AF141901</i> and in exon 25 for <i>SHANK2E</i>, <i>ProSAP1A</i> and <i>ProSAP1</i>. Conserved domains of protein interaction or protein binding site are represented in color: ANK (red), SH3 (orange), PDZ (blue) and SAM (green), H (pink), D, (dark blue) and C (purple). Black stars identify the alternative spliced exons (‘brain-specific exons’ in turquoise: 19, 20 and 23). B. RT-PCRs of <i>SHANK2</i> isoforms on RNA from different human control tissues (Clontech), and different brain regions of four controls (2 males and 2 females). The amplified regions specific to each isoform of <i>SHANK2</i> are indicated by gray boxes. C. Alternative splicing of human <i>SHANK2</i>; exons 19, 20 and 23 are specific to the brain. ANK, ankyrin; SH3, Src homology 3; PDZ, PSD95/DLG/ZO1; SAM, sterile alpha motif; He, heart; Li, liver; B, brain; SM, skeletal muscle; Pl, placenta; K, kidney; Lu, lung; Pa, pancreas; FC, frontal cortex; Hi, hippocampus; TC, temporal cortex; T, thalamus; OC, occipital cortex; Ce, cerebellum; Cx, whole cortex; BLCL, B lymphoblastoid cell lines; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; BSR, brain specific region; H, homer binding site; D, dynamin binding site; C, cortactin binding site. The ages of the two males and the two females studied were 74, 42, 55, and 36 years with a post-mortem interval of 10, 21, 24, and 2 h, respectively.</p

Characterization of CNVs in three patients carrying a <i>de novo</i> deletion of <i>SHANK2</i>.

<p>Paternally or maternally inherited CNVs are indicated by squares and circles, respectively. <i>De novo</i> CNVs are indicated by stars. Deletions and duplications are indicated in red and blue, respectively. CNVs hitting exons or only introns are filled with grey and white, respectively. Squares and circles within star represent <i>de novo</i> CNV of paternal or maternal origin; circles within squares represent CNV inherited by father or mother. ABCC6, ATP-binding cassette, sub-family C, member 6 pseudogene 2; ADAM, ADAM metallopeptidase; AMY1, amylase (salivary); AMY2A, amylase (pancreatic); ARHGAP11B, Rho GTPase activating protein 11B; CAMSAP1L1, calmodulin regulated spectrin-associated protein 1-like 1; CHRNA7, cholinergic receptor, nicotinic, alpha 7; CNTN4, contactin 4; CTNNA3, catenin (cadherin-associated protein), alpha 3; CYFIP1, cytoplasmic FMR1 interacting protein 1; DUSP22, dual specificity phosphatase 22; GALM, galactose mutarotase; GCNT2, glucosaminyl (N-acetyl) transferase 2; GOLGA, golgi autoantigen, golgin subfamily a; GSTT1, glutathione S-transferase theta 1; HLA-DRB, major histocompatibility complex, class II, DR beta; LAMA4, laminin, alpha 4; NIPA, non imprinted in Prader-Willi/Angelman syndrome; NLGN1, neuroligin 1; NME7, non-metastatic cells 7; OR, olfactory receptor; PCDHA, protocadherin alpha; RFPL4B, ret finger protein-like 4B; RHD, Rh blood group, D antigen; SFMBT1, Scm-like with four mbt domains 1; SHANK2, SH3 and multiple ankyrin repeat domains 2; SMC2, structural maintenance of chromosomes 2; TNS3, tensin 3; TUBGCP5, tubulin, gamma complex associated protein 5; UGT2B17, UDP glucuronosyltransferase 2 family, polypeptide B17.</p