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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Wen-Jie WANG,Ning SONG,Li-Sheng LV,Jun-Xia XIE~*,Jun WANG~* Department of Physiology,Shandong Provincial Key Laboratory of Pathogenesis and Prevention of Neurological Disorders and State Key Disciplines:Physiology,Medical College of Qingdao University,Qingdao,266071,China

＜正＞Iron regulatory protein1（IRP1） is considered as a central regulator of cellular iron metabolism,because it regulates the synthesis of proteins that are required for the uptake,storage,and export of iron by binding to mRNA iron-responsive elements（IRE）.It was reported in our lab that both mRNA and protein levels of iron exporter ferroportinl（Fpl） decreased when IRPl was *** activity is modulated by PKC-dependent phosphorylation,possibly phosphorylation state will influence the use of specific IRE-containing *** the IRPl phosphorylation is *** aim of our study was to determine whether phorbol 12-myristate-13- acetate（PMA） affects IRP1 activity and,thus,the levels of Fp1 in the dopaminergic cell line mes23.5 *** viability was assessed by MTT,and IRP1 and Fp1 levels were investigated by Western *** viability of cells treated with 0.01- 2.0μmol/L PMA for 24 h was unchanged compared to that of the *** IRP1 and Fp1 levels were measured after cells were treated with 0.2μmol/L PMA for 24 h.A significant increase of IRP1 protein expression and a decrease of Fp1 levels were observed in the treated cells,which indicated an increased IRP1 activity after PMA *** data suggest that IRP1 can be activated by PKC-dependent phosphorylation in dopaminergic cell line mes23.5 cells.

关键词： mes23.5 cells iron regulatory protein1(IRP1) ferroportin1(Fp1) PKC

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Neurotoxic effect of zinc on mes23.5 dopaminergic cells

Neurotoxic effect of zinc on MES23.5 dopaminergic cells

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

作者： Limin SHI,Jun WANG,Hong JIANG,Jun-Xia XIE~* Department of physiology,Medical college of Qingdao University,Qingdao,266071

Evidence demonstrated that the Zn level in the substantia nigra(SN) of patients with Parkinson’s disease(PD) is high, suggesting that Zn may participate in the pathology of *** the present study,neurotoxic effect of ... 详细信息

关键词： Zn2+ mes23.5 cells Parkinson’s disease Dopaminergic neurons Apoptosis

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Impact of Pitx3 gene knockdown on glial cell line-derived neurotrophic factor transcriptional activity in dopaminergic neurons

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Neural Regeneration Research 2017年第8期12卷 1347-1351页

作者： Jing Chen Xiao-yu Kang Chuan-xi Tang Dian-shuai Gao Experiment Teaching Center of Morphology Xuzhou Medical University Xuzhou Jiangsu Province China Teaching and Research Section of Neurobiology and Anatomy Xuzhou Medical University Xuzhou Jiangsu Province China

Pitx3 is strongly associated with the phenotype, differentiation, and survival of dopaminergic neurons. The relationship between Pitx3 and glial cell line-derived neurotrophic factor（GDNF） in dopaminergic neurons remains poorly understood. The present investigation sought to construct and screen a lentivirus expression plasmid carrying a rat Pitx3 short hairpin（sh）RNA and to assess the impact of Pitx3 gene knockdown on GDNF transcriptional activity in mes23.5 dopaminergic neurons. Three pairs of interference sequences were designed and separately ligated into GV102 expression vectors. These recombinant plasmids were transfected into mes23.5 cells and western blot assays were performed to detect Pitx3 protein expression. Finally, the most effective Pitx3 sh RNA and a dual-luciferase reporter gene plasmid carrying the GDNF promoter region（GDNF-luciferase） were cotransfected into mes23.5 cells. Sequencing showed that the synthesized sequences were identical to the three Pitx3 interference sequences. Inverted fluorescence microscopy revealed that the lentivirus expression plasmids carrying Pitx3-sh RNA had 40-50% transfection efficiency. Western blot assay confirmed that the corresponding Pitx3 of the third knockdown sequence had the lowest expression level. Dual-luciferase reporter gene results showed that the GDNF transcriptional activity in dopaminergic cells cotransfected with both plasmids was decreased compared with those transfected with GDNF-luciferase alone. Together, the results showed that the designed Pitx3-sh RNA interference sequence decreased Pitx3 protein expression, which decreased GDNF transcriptional activity.

关键词： nerve regeneration neurodegeneration Parkinson's disease glial cell line-derived neurotrophic .factor Pitx3 mes23.5 cells shorthairpin RNA gene knockdown plasmid dual-luciferase reporter gene neural regeneration

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