PtdIns (3,4,5) P3 Recruitment of Myo10 Is Essential for Axon Development

Myosin X (Myo10) with pleckstrin homology (PH) domains is a motor protein acting in filopodium initiation and extension. However, its potential role has not been fully understood, especially in neuronal development. In the present study the preferential accumulation of Myo10 in axon tips has been revealed in primary culture of hippocampal neurons with the aid of immunofluorescence from anti-Myo10 antibody in combination with anti-Tuj1 antibody as specific marker. Knocking down Myo10 gene transcription impaired outgrowth of axon with loss of Tau-1-positive phenotype. Interestingly, inhibition of actin polymerization by cytochalasin D rescued the defect of axon outgrowth. Furthermore, ectopic expression of Myo10 with enhanced green fluorescence protein (EGFP) labeled Myo10 mutants induced multiple axon-like neurites in a motor-independent way. Mechanism studies demonstrated that the recruitment of Myo10 through its PH domain to phosphatidylinositol (3,4,5)-trisphosphate (PtdIns (3,4,5) P3) was essential for axon formation. In addition, in vivo studies confirmed that Myo10 was required for neuronal morphological transition during radial neuronal migration in the developmental neocortex

Down-Regulation of Neogenin Accelerated Glioma Progression through Promoter Methylation and Its Overexpression in SHG-44 Induced Apoptosis

Dependence receptors have been proved to act as tumor suppressors in tumorigenesis. Neogenin, a DCC homologue, well known for its fundamental role in axon guidance and cellular differentiation, is also a dependence receptor functioning to control apoptosis. However, loss of neogenin has been reported in several kinds of cancers, but its role in glioma remains to be further investigated.Western blot analysis showed that neogenin level was lower in glioma tissues than in their matching surrounding non-neoplastic tissues (n = 13, p<0.01). By immunohistochemical analysis of 69 primary and 16 paired initial and recurrent glioma sections, we found that the loss of neogenin did not only correlate negatively with glioma malignancy (n = 69, p<0.01), but also glioma recurrence (n = 16, p<0.05). Kaplan-Meier plot and Cox proportional hazards modelling showed that over-expressive neogenin could prolong the tumor latency (n = 69, p<0.001, 1187.6 ± 162.6 days versus 687.4 ± 254.2 days) and restrain high-grade glioma development (n = 69, p<0.01, HR: 0.264, 95% CI: 0.102 to 0.687). By Methylation specific polymerase chain reaction (MSP), we reported that neogenin promoter was methylated in 31.0% (9/29) gliomas, but absent in 3 kinds of glioma cell lines. Interestingly, the prevalence of methylation in high-grade gliomas was higher than low-grade gliomas and non-neoplastic brain tissues (n = 33, p<0.05) and overall methylation rate increased as glioma malignancy advanced. Furthermore, when cells were over-expressed by neogenin, the apoptotic rate in SHG-44 was increased to 39.7% compared with 8.1% in the blank control (p<0.01) and 9.3% in the negative control (p<0.01).These observations recapitulated the proposed role of neogenin as a tumor suppressor in gliomas and we suggest its down-regulation owing to promoter methylation is a selective advantage for glioma genesis, progression and recurrence. Furthermore, the induction of apoptosis in SHG-44 cells after overexpression of neogenin, indicated that neogenin could be a novel target for glioma therapy

Effect of Yoga Intervention on Problem Behavior and Motor Coordination in Children with Autism

Children with autism exhibit more pronounced symptoms of both problem behaviors and motor coordination difficulties. Yoga, recognized as an effective intervention modality, can be valuable after assessing its efficacy in addressing problem behaviors and motor coordination challenges, ultimately contributing to symptom alleviation in autism. The randomized controlled trial (RCT) was used to divide 17 children with autism into an intervention group (n = 9) and a control group (n = 8). The intervention group participated in an 8-week yoga intervention training (three sessions/week, 45–50 min/session), and the control group did not participate in yoga training but only in daily program activities. Pre-test, mid-test, post-test, and after delayed test, teachers assessed the effect of yoga intervention on problem behaviors of children with autism through the Aberrant Behavior Checklist (ABC) and the effect of yoga intervention on motor coordination through the Movement Assessment Battery for Children—Second Edition (MABC2). Results show that the yoga intervention is effective in reducing problem behaviors and improving motor coordination in children with autism. Yoga intervention significantly reduces irritability and social withdrawal in children with autism. Yoga intervention had the most significant improvement in ball skills and static and dynamic balance

The 69 patients’ classified statistic according to clinical and pathological features (January, 2006 – January, 2011).

<p>IOD: integral optical density;</p>*<p>: independent <i>t</i>-test,</p>†<p>: One-Way ANOVA. All tumor sizes were measured by CT or MRI except the data from visual inspection in the operation (details in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038074#pone.0038074.s002" target="_blank">Table S2</a>),</p>‡<p>: n = 57. IOD values were supplied as Mean±Standard Deviation.</p

Overexpression of neogenin in SHG-44 cell line.

<p>(A) Western blot results showed neogenin expression in the blank control which was untreated (Con), the negative control which was transfected by empty vector (Vec) and the over-expressive group which was transfected by neogenin expression plasmid (Neo) in SHG-44 cell line at 48 hours after transfection; (B-D) Status of cells in the blank, negative and over-expressive group of neogenin respectively at 48 h after transfection. (E) Apoptotic distribution maps of cells in the blank, negative and over-expressive group were drew by flow cytometry assay respectively. (F) Average apoptotic rate of cells in the blanck, negative and over-expressive group in the flow cytometry assay, One-Way ANOVA, n = 3, **<i>p</i><0.01, error bars indicate standard error means.</p

Immunohistochemical analysis of neogenin in primary gliomas.

<p>(A–D) Representative immunohistochemical pictures of grade I-IV glioma respectively, (A’–D’) Magnification of the red squares in (A–D), (A–D) 200× magnification; (A’–D’) 400× magnification. (E) Scatter diagram of neogenin expression in 69 primary gliomas, blue bars are the means of four grades, n = 69, One-Way ANOVA was used in statistical analysis. (F) Histogram of mean neogenin expression in low-grade gliomas and high-grade gliomas, statistical analysis was performed with independent <i>t</i>-test. *<i>p</i><0.05, **<i>p</i><0.01, error bars indicate standard error means.</p

Multivariate hazard ratios of progression to high-grade gliomas in 69 primary glioma patients (January, 2006 – January, 2011).

<p>Multivariate hazard ratios and 95% Confidence Intervals were obtained by using Latency as time-scale and High-grade glioma as Failure event. ^‡: divided by Median of neogenin IODs listed in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038074#pone.0038074.s002" target="_blank">Table S2</a>.</p

Clinicopathologic features and relative neogenin level profile in 13 paired surrounding and glioma tissues (January – November, 2011).

<p>SD: standard deviation;</p>*<p>: paired <i>t</i>-test.</p

Kaplan-Meier survival curves for overall progression to high-grade glioma.

<p>Failure event for computation of this curve was diagnosed as the high-grade glioma. Higher neogenin patients are marked in green color, lower neogenin patients are marked in blue color.</p

Primary and recurrent clinicopathologic features of 16 patients (January, 2001 – January, 2011).

<p>IOD: integral optical density;</p><p>IOD values were supplied as Mean±Standard Deviation.</p>*<p>: paired <i>t</i>-test;</p>†<p>: undefined grade gliomas.</p