Dai Zhang

Positive Association of the Oxytocin Receptor Gene (OXTR) with Autism in the Chinese Han Population

BACKGROUND Previous research has suggested that the social impairments exhibited by individuals with autism are associated with changes in plasma oxytocin (OT) levels. The physiologic effects of oxytocin are mediated through its specific receptors (OTRs), and numerous studies have implicated OTRs in the regulation of social cognition and behavior. Animal models and linkage data from genome screens indicate that the oxytocin receptor gene (OXTR) is an excellent candidate for research concerning psychiatric disorders, particularly those involving social impairments, such as autism. METHODS We genotyped four single nucleotide polymorphisms (SNPs) located within the OXTR gene of 195 Chinese Han autism trios, using polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS The family-based association test (FBAT) revealed a significant genetic association between autism and two of the SNPs tested (rs2254298 A: Z = 2.287, p = .0222; rs53576 A: Z = 2.573, p = .0101). When haplotypes were constructed with two, three, and four markers, the haplotype-specific FBAT revealed that a number of haplotypes, particularly those involving rs53576, were significantly associated with autism. Furthermore, haplotypes constructed with all markers showed a significant excess transmission for the specific and global haplotype analyses (p = .0020 and .0289, respectively). CONCLUSIONS These data suggest an involvement of OXTR in the susceptibility to autism, and replication is important.

MirSNP, a database of polymorphisms altering miRNA target sites, identifies miRNA-related SNPs in GWAS SNPs and eQTLs

BackgroundNumerous single nucleotide polymorphisms (SNPs) associated with complex diseases have been identified by genome-wide association studies (GWAS) and expression quantitative trait loci (eQTLs) studies. However, few of these SNPs have explicit biological functions. Recent studies indicated that the SNPs within the 3’UTR regions of susceptibility genes could affect complex traits/diseases by affecting the function of miRNAs. These 3’UTR SNPs are functional candidates and therefore of interest to GWAS and eQTL researchers.DescriptionWe developed a publicly available online database, MirSNP (http://cmbi.bjmu.edu.cn/mirsnp), which is a collection of human SNPs in predicted miRNA-mRNA binding sites. We identified 414,510 SNPs that might affect miRNA-mRNA binding. Annotations were added to these SNPs to predict whether a SNP within the target site would decrease/break or enhance/create an miRNA-mRNA binding site. By applying MirSNP database to three brain eQTL data sets, we identified four unreported SNPs (rs3087822, rs13042, rs1058381, and rs1058398), which might affect miRNA binding and thus affect the expression of their host genes in the brain. We also applied the MirSNP database to our GWAS for schizophrenia: seven predicted miRNA-related SNPs (p < 0.0001) were found in the schizophrenia GWAS. Our findings identified the possible functions of these SNP loci, and provide the basis for subsequent functional research.ConclusionMirSNP could identify the putative miRNA-related SNPs from GWAS and eQTLs researches and provide the direction for subsequent functional researches.

Family‐based association study between autism and glutamate receptor 6 gene in Chinese Han trios

The glutamate pathways are involved in diverse processes such as learning and memory, epilepsy, and they play important roles in neural plasticity, neural development, and neurodegeneration. It has been proposed that autism could be a hypoglutamatergic disorder. Recently, Jamain et al. [Mol Psychiatry 7:302–310] reported that the glutamate receptor 6 (GluR6 or GRIK2) is in linkage disequilibrium with autism. In the present study, the transmission disequilibrium test (TDT) and the haplotype transmission were performed to analyze the four SNPs (SNP1: rs995640; SNP2: rs2227281; SNP3: rs2227283; SNP4: rs2235076) of GluR6 in 174 Chinese Han parent‐offspring trios. The TDT demonstrated that the two SNPs (SNP2 and SNP3) showed preferential transmission (TDT P = 0.032). The global χ2 test for haplotype transmission also revealed an association between GluR6 and autism (χ2 = 10.78, df = 3, P = 0.013). Our results suggested that GluR6 is in linkage disequilibrium with autism.

Positive association of the Disrupted‐in‐Schizophrenia‐1 gene (DISC1) with schizophrenia in the Chinese han population

Disrupted‐in‐Schizophrenia‐1 (DISC1) is located on 1q42.1, one of the most promising susceptibility loci in schizophrenia linkage studies. A non‐synonymous genetic variation rs821616 (Ser704Cys) in DISC1, has recently been shown to be associated with schizophrenia in family‐based study [Callicott et al. (2005); Proc Natl Acad Sci USA 102: 8627–8632]. In order to further confirm this issue, we examined four single nucleotide polymorphisms (SNPs) in a chromosomal region spanning 42 kb of this gene, namely rs821616, rs821597, rs4658971, and rs843979, in Chinese sample of 313 schizophrenia patients and 317 healthy controls. Our results showed that two SNPs had strong associations with schizophrenia (rs821616: Allele A > T, χ2 = 7.8006, df = 1, P = 0.0052; Genotype, χ2 = 7.7935, df = 2, P = 0.0203; rs821597: Allele A > G, χ2 = 9.5404, df = 1, P = 0.0020; Genotype, χ2 = 12.2780, df = 2, P = 0.0022). When haplotypes were constructed with two, three, and four markers, a number of haplotype combinations, especially those including rs821616 and rs821597, were significantly associated with schizophrenia. Furthermore, there was a strong evidence for association in a four‐marker haplotype analysis (χ2 = 7.686, df = 4, P = 0.005581, corrected P = 0.006199). Although the case‐control and family‐based association studies both suggest that DISC1 gene may play a role in genetic susceptibility to schizophrenia, the risk haplotypic combinations have subtle differences in the two studies. Our findings provide further evidence for DISC1 as a predisposing gene involved in schizophrenia in the Chinese Han Population.

Tenuigenin treatment decreases secretion of the Alzheimer’s disease amyloid β-protein in cultured cells

Amyloid beta-protein (A beta) is a pivotal pathological factor in Alzheimers disease (AD). Tenuigenin, extracted from the Chinese herb Polygala tenuifolia, seems to ameliorate the reduction in cholinergic function on rat models induced by A beta. To examine this therapeutic effect, we tested whether Tenuigenin could inhibit secretion of A beta in neuroblastoma cells stably transfected with two amyloid precursor protein (APP) constructs: the APP695 cDNA (SH-SY5Y APP695) and the C-terminal 99 amino acid residues of APP plus the signal peptide (SH-SY5Y SPA4CT). Tenuigenin inhibited the secretion of A beta and the C-terminal 99 amino acids of APP (C99) in SH-SY5Y APP695 cells, but did not change the A beta and C99 levels in SH-SY5Y SPA4CT cells. Fluorescence Resonance Energy Transfer (FRET) assays showed that Tenuigenin inhibited the proteolytic activities of BACE1 (beta-secretase) on its substrate in vitro. In addition, Tenuigenin did not demonstrate any cytotoxic effects, nor did it affect APP mRNA expression, holoAPP synthesis or sAPP alpha secretion. Our data suggest that Tenuigenin can inhibit the secretion of A beta in SH-SY5Y APP 695 cells via BACE1 inhibition. Taken together, these results suggest that Tenuigenin may be worthy of future study as an anti-AD drug.

A diffusion tensor imaging study of middle and superior cerebellar peduncle in male patients with schizophrenia.

Many studies have confirmed that the cerebellum takes part in higher-order cognitive coordination; profound fibers projecting to and from the cerebellum underlie its cognitive function. Since the superior and middle cerebellar peduncles are the main pathways of neural fibers in the cerebellum, these structures became the focus of our interest in evaluating the cognitive dysfunction reported in schizophrenia. Diffusion tensor imaging (DTI) was used to examine the anatomical integrity of the neural fibers in the superior and middle cerebellar peduncles. DTI was performed on 29 patients and 20 normal controls; we subsequently calculated the fractional anisotropy and mean diffusivity in these regions. Statistical analysis revealed that there was no significant difference between patients with schizophrenia and our matched control group. No structural abnormalities were detected in the white matter of the superior and middle cerebellar peduncles.

Association between the FOXP2 gene and autistic disorder in Chinese population

Several genomewide screens indicated that chromosome 7q was linked to autistic disorder. FOXP2, located on 7q31, is a putative transcription factor containing a polyglutamine tract and a forkhead DNA binding domain. It is one member of the forkhead family who are known to be key regulators of embryogenesis. A point mutation at a highly conserved residue within the forkhead domain co‐segregated with affected status in the KE family who was a unique three generation pedigree with a severe speech and language disorder and FOXP2 was directly disrupted by a translocation in an individual who had similar deficits as those of the KE family. Several studies have investigated the role of FOXP2 polymorphisms in autism and none of them found positive association. We performed a family‐based association study of three single nucleotide polymorphisms (SNPs) of FOXP2 in 181 Chinese Han trios using the analyses of transmission/disequilibrium test (TDT) and haplotype. We found a significant association between autistic disorder and one SNP, as well as with specific haplotypes formed by this SNP with two other SNPs we investigated. Our findings suggest that the FOXP2 gene may be involved in the pathogenesis of autism in Chinese population.

Association of the ENGRAILED 2 (EN2) gene with autism in Chinese Han population

Human ENGRAILED 2 (EN2) gene is localized to 7q36, an autism susceptibility locus. En2 knockout mice display hypoplasia of cerebellum and a decrease in the number of Purkinje cell, which are similar to those reported for individuals with autism. Furthermore, deficits in social behavior were detected in En2−/− mice. Two recent studies have demonstrated that two intronic SNPs (rs1861972, rs1861973) in the EN2 gene are significantly associated with autism. To investigate whether this finding could be replicated in Chinese Han population, we performed the association study between eight single nucleotide polymorphisms (SNPs) of the EN2 gene and autism in 210 Chinese Han trios, using the family‐based association test (FBAT). The present study demonstrated that a preferential transmission of the rs3824068 A‐allele to affected offspring (A > G: Z = 2.399, P = 0.0165). After the Bonferroni correction, this statistical significance of preferential transmission did not remain. However, when haplotypes were constructed with multiple markers, a number of haplotypes including three two‐marker haplotypes, nine three‐marker haplotypes, one four‐marker haplotype, and one six‐marker haplotype, all of which contain the major allele A of rs3824068, displayed significantly associated with autism. These results were still significant after using the permutation method to obtain empirical P values. Thus, our data provide evidence that the EN2 gene may be implicated in the predisposition to autism in the Chinese Han population.

Hemispheric asymmetry in cognitive division of anterior cingulate cortex: A resting-state functional connectivity study

The cognitive division of anterior cingulate cortex (ACC-cd) plays an important role in cognitive control via a distributed attention network. The structural hemispheric asymmetries of ACC have been revealed by several neuroimaging studies. However potential functional hemispheric asymmetries of ACC remain less clear. Investigating the functional hemispheric asymmetries of ACC helps for a better understanding of ACC function. The aim of this study was to use resting-state functional magnetic resonance imaging (fMRI) to examine hemispheric differences in the functional networks associated with ACC-cd in the two hemispheres. ROI-based functional connectivity analysis was performed on a group of 49 right-handed healthy volunteers. The left and right ACC-cd showed significant differences in their patterns of connectivity with a variety of brain regions, including the dorsolateral prefrontal cortex, inferior parietal lobule, superior parietal lobule and dorsal posterior cingulate cortex in their ipsilateral cerebral cortex, as well as cerebellar tonsil and inferior semilunar lobule in their contralateral cerebellar hemisphere. Specifically, for these areas, we found significantly greater connectivity strength with ACC-cd in the right hemisphere than the left, regardless of whether the connection was positive or negative. The current results highlight the presence of clear asymmetries in functional networks associated with ACC-cd. Future functional imaging studies are needed to give greater attention to the lateralized ACC functional networks which are observed.

Quetiapine enhances oligodendrocyte regeneration and myelin repair after cuprizone-induced demyelination

Myelin and oligodendrocyte dysfunctions have been consistently found in patients with schizophrenia. The effect of antipsychotics on myelin disturbances is unknown. The present study examined the effects of quetiapine on oligodendrocyte regeneration and myelin repair in a demyelination animal model. C57BL/6 mice were fed with cuprizone (0.2% w/w) for 12 weeks to induce chronic demyelination and oligodendrocyte degeneration, after which cuprizone was withdrawn to allow recovery. Quetiapine (10mg/kg/day) or vehicle (water) was administrated orally to mice for 0, 2, 3, or 4 weeks after cuprizone withdrawal. Locomotor activity and Y-maze tests were used to evaluate behavioral changes in the mice. Immunohistochemical staining was used to detect morphological and biological changes in the brains. Cuprizone administration for 12 weeks resulted in severe demyelination, locomotor hyperactivity, and working memory impairment in mice. Remyelination occurred when cuprizone was withdrawn. Quetiapine treatment during the recovery period significantly improved the spatial working memory and increased myelin restoration. Quetiapine treatment also enhanced the repopulation of mature oligodendrocytes in the demyelinated lesions, which was associated with down-regulation of transcription factor olig2 in the process of cell maturation. The results of this study demonstrated that quetiapine treatment during the recovery period improves spatial working memory and promotes oligodendrocyte development and remyelination. This study supports the role of oligodendrocyte dysfunction in memory deficits in a schizophrenia mouse model and suggests that quetiapine may target oligodendrocytes and improve cognitive function.