MicroRNA-218 regulates neuronal radial migration and morphogenesis by targeting Satb2 in developing neocortex.

Neuronal migration and morphogenesis are fundamental processes for cortical development. Their defects may cause abnormities in neural circuit formation and even neuropsychiatric disorders. Many proteins, especially layer-specific transcription factors and adhesion molecules, have been reported to regulate the processes. However, the involvement of non-coding RNAs in cortical development has not been extensively studied. Here, we identified microRNA-218 (miR-218) as a layer V-specific microRNA in mouse brains. Expression of miR-218 was elevated in patients with autism spectrum disorder (ASD) and schizophrenia. We found in this study that miR-218 overexpression in developing mouse cortex led to severe defects in radial migration, morphogenesis, and spatial distribution of the cortical neurons. Moreover, we identified Satb2, an upper-layer marker, as a molecular target repressed by miR-218. These results suggest an underlying mechanism of miR-218 involvement in neuropsychiatric disorders, and the interactions of layer-specific non-coding RNAs and proteins in regulating cortical development.

Inhibition of Autism-Related Crm1 Disrupts Mitosis and Induces Apoptosis of the Cortical Neural Progenitors.

De novo microdeletion of chromosome 2p15-16.1 presents clinically recognizable phenotypes that include mental retardation, autism, and microcephaly. Chromosomal maintenance 1 (CRM1) is a gene commonly missing in patients with 2p15-16.1 microdeletion and one of two genes found in the smallest deletion case. In this study, we investigate the role and mechanism of Crm1 in the developing mouse brain by inhibiting the protein or knocking down the gene in vivo. Inhibition of Crm1 reduces the proliferation and increases p53-dependent apoptosis of the cortical neural progenitors, thereby impeding the growth of embryonic cerebral cortex. Live imaging of mitosis in ex vivo embryonic brain slices reveals that inhibition of CRM1 arrests the cortical progenitors at metaphase. The arrested cells eventually slip into a pseudo-G1 phase without chromosome segregation. The mitotic slippage cells are marked by persistent expression of the spindle assembly checkpoint (SAC), repressing of which rescues the cells from apoptosis. Our study reveals that activating the SAC and inducing the mitotic slippage may lead to apoptosis of the cortical neural progenitors. The resulting cell death may well contribute to microcephaly associated with microdeletion of chromosome 2p15-16.1 involving CRM1.

Transient Expression of Fez Family Zinc Finger 2 Protein Regulates the Brn3b Gene in Developing Retinal Ganglion Cells.

Retinal ganglion cells (RGCs) are projection neurons in the neural retina that relay visual information from the environment to the central nervous system. The early expression of MATH5 endows the post-mitotic precursors with RGC competence and leads to the activation ofBrn3bthat marks committed RGCs. Nevertheless, this fate commitment process and, specifically, regulation ofBrn3bremain elusive. To explore the molecular mechanisms underlying RGC generation in the mouse retina, we analyzed the expression and function of Fez family zinc finger 2 (FEZF2), a transcription factor critical for the development of projection neurons in the cerebral cortex.Fezf2mRNA and protein were transiently expressed at embryonic day 16.5 in the inner neuroblast layer and the prospective ganglion cell layer of the retina, respectively. Knockout ofFezf2in the developing retina reduced BRN3B+ cells and increased apoptotic cell markers.Fezf2knockdown by retinalin uteroelectroporation diminished BRN3B but not the coexpressed ISLET1 and BRN3A, indicating that the BRN3B decrease was the cause, not the result, of the overall reduction of BRN3B+ RGCs in theFezf2knockout retina. Moreover, the mRNA and promoter activity ofBrn3bwere increasedin vitroby FEZF2, which bound to a 5' regulatory fragment in theBrn3bgenomic locus. These results indicate that transient expression ofFezf2in the retina modulates the transcription ofBrn3band the survival of RGCs. This study improves our understanding of the transcriptional cascade required for the specification of RGCs and provides novel insights into the molecular basis of retinal development.

Magnetic Resonance Features of Acquired Immune Deficiency Syndrome Involving Central Nervous System Diseases by Intelligent Fuzzy C-Means Clustering (FCM) Algorithm.

This study was aimed to explore the application of fuzzy C-means (FCM) algorithm in MR images of acquired immune deficiency syndrome (AIDS) patients. Sixty AIDS patients with central nervous disease were selected as the research object. A method of brain MR image segmentation based on FCM clustering optimization was proposed, and FCM was optimized based on the neighborhood pixel correlation of gray difference. The correlation was introduced into the objective function to obtain more accurate pixel membership and segmentation features of the image. The segmented image can retain the original image information. The proposed algorithm can clearly distinguish gray matter from white matter in images. The average time of image segmentation was 0.142 s, the longest time of level set algorithm was 2.887 s, and the running time of multithreshold algorithm was 1.708 s. FCM algorithm had the shortest running time, and the average time was significantly better than other algorithms (P < 0.05). FCM image segmentation efficiency was above 90%, and patients can clearly display the location of lesions after MRI imaging examination. In summary, FCM algorithm can effectively combine the spatial neighborhood information of the brain image, segment the BRAIN MR image, analyze the characteristics of AIDS patients from different directions, and provide effective treatment for patients.

The Association of OTX1 rs17850223 Polymorphisms in Han Chinese Patients with Idiopathic Epilepsy.

This study is aimed at investigating the association between orthodenticle homeobox 1 (OTX1) gene polymorphisms and idiopathic epilepsy in a cohort of Han Chinese patients. We carried out a case-control study on 147 patients with idiopathic epilepsy and 150 healthy controls. Genomic DNA was isolated from 1 ml of ethylene diamine tetraacetic acid (EDTA)-treated blood. The OTX1 coding sequence was divided into three parts and amplified using PCR, and the products were genotyped using the Sanger sequencing method. All OTX1 coding sequences were conserved except for rs17850223 located on the fifth exon. The frequency of the CC, CG, and GG genotypes showed no statistical differences between the idiopathic epileptic patients and the controls. The rs17850223 G allele distribution was also similar between the idiopathic epileptic patients and the controls. Interestingly, the frequency of the GG genotype was significantly higher in the patients with generalized seizures compared with that of the controls (12.2% vs. 2%, p = 0.012), and a greater distribution of the rs17850223 G allele was also seen in the patients with generalized seizures compared with controls (18.3% vs. 10%, p = 0.049). rs17850223 might play a critical role in Chinese idiopathic epileptic patients with generalized seizure activity.

Primary anorectal malignant melanoma: a case report.

Anorectal melanoma (ARM) is rare and lethal. We report a case of a 48-year-old woman with 9 months of rectal swelling and bleeding. Physical examination revealed a mass about 5 × 6 cm on the anterior wall of the rectum, 3 cm from the anal verge, and the patient underwent abdominoperineal resection (APR). After hematoxylin-eosin staining and immunohistochemical staining, it was considered an ARM, which is an aggressive disease with a poor survival. Immunohistochemical staining showed the tumor to be positive for S-100, Melan A, Ki67 proliferative index of 70%, and negative for HMB45. The melanoma had infiltrated the adventitia and metastasized to the (intestinal) 16/16 lymph nodes with cancerous nodule formation. There were multiple organs with metastasis (liver, spleen, pancreas, lung and subcutaneous soft tissue) three months after operation. Overall, pre-operative biopsy may be insufficient to make a definite diagnosis, and immunohistochemistry is necessary. Therefore, the gold standard treatment for ARM is oncological radical surgical resection.

Curcumin-induced promoter hypermethylation of the mammalian target of rapamycin gene in multiple myeloma cells.

Curcumin, a polyphenol derived from the rhizome of Curcuma, is a potential tumor inhibitor through affecting signaling pathways and epigenetic regulation. The mammalian target of rapamycin (mTOR) gene serves a crucial role in the carcinogenesis of multiple myeloma. The curcumin-induced epigenetic regulation of mTOR, including promoter DNA methylation in multiple myeloma, has not yet been fully elucidated. In the present study, antitumor effects of curcumin were investigated in RPMI-8226 and NCI-H929 cells using an MTT assay and flow cytometry. The expression of mTOR and DNA methyltransferase proteins were determined by western blot analysis, and the methylation status of the mTOR promoter were detected by sequencing following bisulfite conversion. The results of the present study revealed that the half-maximal inhibitory concentration of curcumin was 10 µM in myeloma cells. Following curcumin treatment, the mRNA and protein expression levels of mTOR were decreased by 43.31 and 39.34% in NCI-H929 cells, respectively. The promoter of mTOR, located in chromosome 1 (chromosome position, 11262153-11263153), contains a CpG island that was hypermethylated in myeloma cells following curcumin treatment. The expression levels of DNA methyltransferase (DNMT)3a and DNMT3b were increased in curcumin-treated cells. Collectively, these results indicate that curcumin serves a role in the epigenetic regulation of mTOR expression, and that mTOR downregulation is due to promoter hypermethylation, which may be associated with DNMT3a and DNMT3b upregulation. The results of the present study contribute towards improving the understanding of curcumin treatment in multiple myeloma and provide novel insights into the molecular mechanisms underlying the epigenetic regulation of mTOR.

The ICAM-1 K469E polymorphism is associated with the risk of coronary artery disease: a meta-analysis.

BACKGROUND: The intercellular adhesion molecule-1 (ICAM-1) K469E polymorphism has been indicated to be correlated with coronary artery disease (CAD) susceptibility, but the results of studies are still debatable. Thus, a meta-analysis was carried out. METHODS: Databases including PubMed, Embase, Web of Science, and CNKI were searched. Data were extracted and pooled odds ratios (OR) with 95% confidence intervals (CI) were calculated. RESULTS: Eighteen studies with 3546 cases and 3852 controls were included in this meta-analysis. The association between the ICAM-1 K469E polymorphism and the risk of CAD was significant (OR=1.77; 95% CI, 1.52-2.05; P<0.01; I=27%). This result remained statistically significant when the adjusted ORs were combined (OR=1.95; 95% CI, 1.78-2.14; P<0.01; I=0%). When stratified by ethnicity, a significantly increased risk was observed in Whites (OR=1.75; 95% CI, 1.37-2.23; P<0.01; I=58%) and in Asians (OR=1.80; 95% CI, 1.45-2.24; P<0.01; I=0%). A significantly increased risk of myocardial infarction was observed (OR=2.24; 95% CI, 1.72-2.92; P<0.01; I=38%). CONCLUSION: In conclusion, this meta-analysis suggested that the ICAM-1 K469E polymorphism was a risk factor for CAD.

Atlastin-1 regulates dendritic morphogenesis in mouse cerebral cortex.

Hereditary spastic paraplegias (HSPs) are human genetic disorders characterized by lower extremity spasticity and weakness. Mutations in atlastin-1 (ATL1) have been identified in patients with HSP SPG3A. However, the function of ATL1 in the mammalian brain remains unclear. Here, we found that expression of ATL1 mRNA was restricted in the deep layer of mouse cerebral cortex during the early development. We examined ATL1 functions by delivering its plasmids to the upper layer cortical neurons using in utero electroporation. The effects of ectopic expression in the pyramidal neurons were determined both in culture and in situ at postnatal stages of neocortical development. In cultured cortical neurons, overexpressing ATL1 increased dendrite growth and arborization, whereas HSP-associated mutant R217Q, which is devoid of GTPase activity, had no such effects. Consistent with this, in vivo expression of wild type ATL1, but not of the mutant R217Q, increased dendritic growth of the cortical neurons. This suggests that the role of ATL1 on dendritic morphogenesis depends on its GTPase activity. The expression of ATL1 and R217Q did not affect the migration of cortical neurons. These results indicate that ATL1 regulates dendritic morphogenesis, which may provide new insights into the neuropathogenic mechanism of hereditary spastic paraplegia SPG3A.

Gene expression and IG-DMR hypomethylation of maternally expressed gene 3 in developing corticospinal neurons.

The mammalian cerebral cortex plays a central role in higher cognitive functions and in the complex task of motor control. Maternally expressed gene 3 (Meg3) appears to play a role in cortical development and neurodegeneration, but the expression and regulation of Meg3 in the cortex is not clear. In this study, we examined the expression of transcript variants of Meg3 in the developing mouse cerebral cortex. By in situ hybridization, we found that a novel transcript variant of Meg3 with 8 small exons was expressed in the developing cortex, whereas the long isoforms of Meg3 (~11 kb) were enriched in corticospinal neurons (CSNs) in layer V of the cortex. No transcript variants of Meg3 were found in the neural progenitors at E12.5, when the intergenic differential methylation region (IG-DMR) near Meg3 was highly methylated. IG-DMR became demethylated at E15.5 and remained hypomethylated in early CSNs isolated from Fezf2-EGFP transgenic mice. The expression of Meg3 transcript variant 1 was inversely correlated with the IG-DMR methylation level during development. Moreover, expression of paternally expressed gene Peg11 was limited to the upper layers, consistent with the idea that the maternally expressed gene may be preferentially transcribed in the lower layers of the cortex. The spatiotemporal expression pattern of Meg3 suggests that it may participate in the early development of CSNs and contribute to cortical malfunctions related to aberrant imprinting in Meg3.

Area dependent expression of ZNF312 in human fetal cerebral cortex.

The human cerebral neocortex is divided into six layers consisting of specific neuronal cell types and connections. To determine the distribution of cortical neurons during early development, we examined the expressions of layer-specific markers in human midterm fetal brains. Layer V marker ZNF312 is expressed in most cortical areas, but not in the prospective somatosensory association area. Expression of layer IV marker RORbeta is also diminished in this region but increased in the primary visual cortex, where expression of ZNF312 is reduced. Our results indicate that ZNF312 and other layer markers have area dependent expressions in the human fetal cortex.