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Effect of chondroitin sulfate proteoglycans on neuronal cell adhesion, spreading and neurite growth in culture

Neural Regeneration Research 2018年第2期13卷 289-297页

作者： Jingyu Jin Sharada Tilve Zhonghai Huang Libing Zhou Herbert M.Geller Panpan Yu Guangdong-Hongkong-Macao Institute of CNS Regeneration Ministry of Education Joint International Research Laboratory of CNS Regeneration Jinan University Laboratory of Developmental Neurobiology Cell Biology and Physiology Center National Heart Lung and Blood Institute National Institutes of Health

As one major component of extracellular matrix （ECM） in the central nervous system, chondroitin sul- fate proteoglycans （CSPGs） have long been known as inhibitors enriched in the glial scar that prevent axon regeneration after injury. Although many studies have shown that CSPGs inhibited neurite out- growth in vitro using different types of neurons, the mechanism by which CSPGs inhibit axonal growth remains poorly understood. Using cerebellar granule neuron （CGN） culture, in this study, we evaluated the effects of different concentrations of both immobilized and soluble CSPGs on neuronal growth, in- cluding cell adhesion, spreading and neurite growth. neurite length decreased while CSPGs concentration arised, meanwhile, a decrease in cell density accompanied by an increase in cell aggregates formation was observed. Soluble CSPGs also showed an inhibition on neurite outgrowth, but it required a higher concen- tration to induce cell aggregates formation than coated CSPGs. We also found that growth cone size was significantly reduced on CSPGs and neuronal cell spreading was restrained by CSPGs, attributing to an inhibition on lamellipodial extension. The effect of CSPGs on neuron adhesion was further evidenced by interference reflection microscopy （IRM） which directly demonstrated that both CGNs and cerebral cortical neurons were more loosely adherent to a CSPG substrate. These data demonstrate that CSPGs have an effect on cell adhesion and spreading in addition to neurite outgrowth.

关键词： chondroitin sulfate proteoglycans cell adhesion neurite growth interference reflection microscopy neural regeneration

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Spastin interacts with collapsin response mediator protein 3 to regulate neurite growth and branching

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Neural Regeneration Research 2021年第12期16卷 2549-2556页

作者： Zhi-Sheng Ji Jian-Ping Li Chao-Hua Fu Jian-Xian Luo Hua Yang Guo-Wei Zhang Wutian Wu Hong-Sheng Lin Department of Orthopedics The First Affiliated Hospital of Jinan UniversityGuangzhouGuangdong ProvinceChina Guangdong-Hong Kong-Macao Institute of CNS Regeneration Jinan UniversityGuangzhouGuangdong ProvinceChina Department of Orthopedics Jiangmen Hospital of Sun Yat-sen UniversityJiangmenGuangdong ProvinceChina Re-Stem Biotechnology Co. Ltd.SuzhouJiangsu ProvinceChina Spine Surgery The Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouGuangdong ProvinceChina

Cytoskeletal microtubule rearrangement and movement are crucial in the repair of spinal cord *** plays an important role in the regulation of microtubule *** spastin and collapsin response mediator proteins can regulate neurite growth and branching;however,whether spastin interacts with collapsin response mediator protein 3(CRMP3)during this process remains unclear,as is the mechanism by which CRMP3 participates in the repair of spinal cord *** this study,we used a proteomics approach to identify key proteins associated with spinal cord injury *** then employed liquid chromatography-mass spectrometry to identify proteins that were able to interact with glutathione ***,co-immunoprecipitation and staining approaches were used to evaluate potential interactions between spastin and ***,we co-transfected primary hippocampal neurons with CRMP3 and spastin to evaluate their role in neurite *** spectrometry identified the role of CRMP3 in the spinal cord injury repair *** chromatography-mass spectrometry pulldown assays identified three CRMP3 peptides that were able to interact with ***3 and spastin were co-expressed in the spinal cord and were able to interact with one another in vitro and in ***,CRMP3 overexpression was able to enhance the ability of spastin to promote neurite growth and ***,our results confirm that spastin and CRMP3 play roles in spinal cord injury repair by regulating neurite growth and *** proteins may therefore be novel targets for spinal cord injury *** Institutional Animal Care and Use Committee of Jinan University,China approved this study(approval ***-20181008-03)on October 8,2018.

关键词： collapsin response mediator protein 3 liquid chromatography-mass spectrometry microtubule neurite growth protein interactions proteomics spastin spinal cord injury

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IPP5 inhibits neurite growth in primary sensory neurons by main-taining TGF-β/Smad signaling

IPP5 inhibits neurite growth in primary sensory neurons by m...

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中国神经科学学会第十届全国学术会议

作者： Qing-Jian Han Nan-Nan Gao Guo-Qiang Ma Zhen-Ning Zhang Wen-Hui Yu Jing Pan Qiong Wang Xu Zhang Lan Bao State Key Laboratory of Cell Biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of Sciences Institute of Neuroscience and State Key Laboratory of Neuroscience Shanghai Institutes for Biological SciencesChinese Academy of Sciences

Objective neurite growth is a critical step for successful nerve regeneration after injury and it is regulated by both intrinsic molecules and extracellular *** found that inhibitor 5 of protein phosphatase 1(IPP5),a newly identified inhibitory subunit of protein phosphatase 1(PP1),was highly expressed in rat primary sensory neurons localized in the dorsal root ganglion(DRG),specifically in small primary sensory neurons,and was downregulated after sciatic nerve axotomy,and may serve as a neurite growth inhibitory molecule in primary sensory *** we explore the regulatory functions of IPP5 in DRG neurite ***(1) Knocking down IPP5 with specific shRNA and examining the neurite growth of cultured DRG neurons;(2) Mutating the PP1 binding motif or the PP1inhibiting motif at Thr to determine whether the inhibition of PP1 is responsible for the effect of IPP5 on neurite growth;(3) Blocking TGF-p signaling with TβRI inhibitor SB431542 or Smad2 shRNA to clarify the involvement of TGF-p signaling ***(1) Knocking down IPP5 promote neurite growth in cultured primary sensory neurons.(2) The interaction with and inhibition of PP1 is responsible for the effect of IPP5 on neurite growth.(3) IPP5inhibits neurite growth by regulating the TGF-p signaling *** IPP5 inhibits neurite growth in primary sensory neurons in a PP1-dependent manner and functions by maintaining TGF-p/Smad signaling pathway.

关键词： IPP5 neurite growth PP1 TGF-β/Smad signaling DRG neuron

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CircRtn4 Acts as the Sponge of miR-24-3p to Promote neurite growth

CircRtn4 Acts as the Sponge of miR-24-3p to Promote Neurite ...

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中国生物化学与分子生物学会2020全国学术在线会议

作者： Yue Qi Jun Wan Shenzhen Key Laboratory for Neuronal Structural Biology Biomedical Research InstituteShenzhen Peking University-The Hong Kong University of Science and Technology Medical Center

Circular RNAs(circRNAs) are covalently closed single-stranded RNA molecules derived from precursor mRNA(pre-mRNA)back-splicing and play an important role in many diseases or life *** circRNAs are usually expressed at relatively low levels,recent studies have showed that some of them are enriched in the mammalian brain,and many of their functions remain *** expressed CircRtn4 in the mouse brain has been reported that the expression increased significantly with the differentiation of *** our study,knockdown of circRtn4 inhibited neurite growth,while overexpression of circRtn4 significantly increased neurite *** performing Dual-luciferase reporter assay,we identified miR-24-3 p as the circRtn4-associated *** circRtn4,the knockdown of miR-24-3 p increased neurite length,while overexpression of miR-24-3 p significantly inhibited neurite *** conclusion,circRtn4 may promote neurite growth by acting as miR-24-3 p sponge.

关键词： CircRtn4 MiR-24-3p neurite growth

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Midkine promotes perineural invasion in human pancreatic cancer

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World Journal of Gastroenterology 2014年第11期20卷 3018-3024页

作者： Jun Yao Wen-Yao Li Shuo-Guo Li Xiao-Shan Feng She-Gan Gao Department of Oncology First Affiliated HospitalHenan University of Science and Technology

AIM: To investigate midkine (MK) and syndecan-3 protein expression in pancreatic cancer by immunohistochemistry, and to analyze their correlation with clinicopathological features, perineural invasion, and prognosis.

关键词： Midkine Syndecan-3 Pancreatic cancer neurite growth Perineural invasion

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Transplantation of NT-3 delivery acellular optic nerve scaffold to improve motor function in transected rat spinal cord

Transplantation of NT-3 delivery acellular optic nerve scaff...

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中国神经科学学会第十二届全国学术会议

作者： Ge Li Jia-Hui Sun Yuan-Shan Zeng Key Laboratory for Stem Cells and Tissue Engineering (Sun Yat-sen University) Ministry of Education Department of Histology and Embryology Zhongshan School of Medicine Sun Yat-sen University

Objective To generate a bioactive scaffold with line-shaped tubes in vitro and investigate whether the scaffold transplanted into the injury site could promote nerve fiber regeneration and functional recovery in rat spinal cord transected completely. Methods By using aseptic decellular, DNAase hydrolysis and freeze-drying techniques, natural optic nerve was removed all cell component and coated with neurotrophin-3（NT-3）/fibroin complex at the surface to fabricate a NT-3 delivery acellular optic nerve scaffold. Then, dorsal root ganglion（DRG） was seeded on the surface of NT-3 delivery acellular optic nerve scaffold in vitro to observe the effect of the scaffold on DRG neurite growth, and the scaffold were transplanted into the injury site of spinal cord injury of rat. Results Fresh normal optic nerve of longitudinal section was stained by HE showing high-density tubes with optic nerve fibers. By using aseptic decellular technique and freeze-drying technique, the NT-3 delivery acellular optic nerve scaffold still maintained its natural 3 dimension（3 D） structure of extracellular matrix. Circle-shaped pipelines made of perineurium-like extracellular matrix in optic nerve of transverse section could be obseved and there were line-shaped tubes with interconnections in optic nerve of longitudinal section after all cell component was removed. These results were further conformed under a scanning electron microscope. The present study proved that cell component had been removed by the results below;*** nuclei were invisible by Hoechst33342 staining. 2. DNA electrophoresis showed that residual DNA fragments were less than 200 bp. 3. DNA quantification showed ＜ 50 ng per mg scaffold dry weight. Therefore, cells seeded in the scaffold could grow without nucleic acid residue effect. Then, after DRG was seeded on the scaffold in vitro, we found that DRG derived NF200 positive neurites could grow a long distance in the surface of the scaffold. Furthermore, two months later aft

关键词： Neutrophin-3 delivery Acellular optic nerve neurite growth Nerve fiber regeneration Spinal cord injury

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Loss of Ahi1 affects early development by impairing BM88/Cend1-mediated neuronal differentiation

Loss of Ahi1 affects early development by impairing BM88/Cen...

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中国神经科学学会第十届全国学术会议

作者： Ling WENG Yung-Feng LIN Alina L.LI Chuan-En WANG Sen YAN Miao SUN Marta A.GAERTIG Naureen MITHA Jun KOSAKA Taketoshi WAKABAYASHI Xing-Shun XU Bei-Sha TANG Shi-Hua LI Xiao-Jiang LI Department of Neurology Xiangya HospitalCentral South University Department of Human Genetics Emory University School of Medicine School of Medical Laboratory Science and Biotechnology Taipei Medical University Department of Cytology and Histology Okayama University Graduate School of Medicine Okayama University Graduate School of Medicine Dentistry and Pharmaceutical Sciences Kansai Medical University Institute of Neuroscience SoochowUniversity State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of SciencesChaoyang District

Mutations in the Abelson helper integration site-1（AHIl） gene result in N-terminal Ahi1 fragments and cause Joubert syndrome,an autosomal recessive brain malformation disorder associated with delayed *** AHIl mutations lead to delayed development remains *** we report that full-length,but not N-terminal,Ahi1 binds Hapl,a huntingtin-associated protein that is essential for the postnatal survival of mice and that tMs binding is regulated during neuronal differentiation by nerve growth *** growth factor induces dephosphorylation of HaplA and decreases its association with Ahil,correlating with increased HaplA distribution in neurite ***,Ahil associates with phosphorylated HaplA in cytosolic,but not in synaptosomal,fractions isolated from mouse brain,suggesting that Ahil functions mainly in the soma of *** spectrometry analysis of cytosolic Ahil immunoprecipitates reveals that Ahil also binds Cendl（cell cycle exit and neuronal differentiation protein1）/BM88,a neuronal protein that mediates neuronal differentiation and is highly expressed in postnatal mouse *** of Ahil reduces the levels of Cendl in the hypothalamus of Ahil KO mice,which show retarded growth during postnatal *** Ahil can stabilize Cendl in cultured ***,overexpression of Cendl can rescue the neurite extension defects of hypothalamic neurons from Ahil KO *** findings suggest that Cendl is involved in Ahil-associated hypothalamic neuronal differentiation in early development,giving us fresh insight into the mechanism behind the delayed development in Joubert syndrome.

关键词： Joubert syndrome Ahi1 Hap1 Cend1 neurite growth

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