title: NCAM-140 Translocation into Lipid Rafts Mediates the Neuroprotective Effects of GDNF.
abstract: Glial cell line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor for substantia nigra dopaminergic (DA) neuronal cells. Recent studies have demonstrated that neural cell adhesion molecule functions as a signal transduction receptor for GDNF. The purpose of this study is to reveal whether neural cell adhesion molecule (NCAM) mediates the protective effects of GDNF on DA neuronal cells and further explore the mechanisms involved. We utilized SH-SY5Y cell line to establish a model of 6-hydroxydopamine (6-OHDA)-injured DA neuronal cells. Lentiviral vectors were constructed to knockdown or overexpress NCAM-140, and a density gradient centrifugation method was employed to separate membrane lipid rafts. 3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), flow cytometric analysis, and western blotting were used to evaluate the protective effects of GDNF. The results showed that GDNF could protect 6-OHDA-injured SH-SY5Y cells via improving cell viability and decreasing the cell death rate and cleaved caspase-3 expression. NCAM-140 knockdown decreased cell viability and increased the cell death rate and cleaved caspase-3 expression, while its overexpression had the opposite effects. Notably, the amount of NCAM-140 located in lipid rafts increased after GDNF treatment. Pretreatment with 2-bromopalmitate, a specific inhibitor of protein palmitoylation, suppressed NCAM-140 translocation to lipid rafts and reduced the NCAM-mediated protective effects of GDNF on injured DA neuronal cells. Our results suggest that GDNF have the protective effects on injured DA cells by influencing NCAM-140 translocation into lipid rafts.
authors.affiliation: Department of Pathophysiology, Xuzhou Medical College, Xuzhou, China.|Department of Neurobiology and Human Anatomy, Xuzhou Medical College, Tongshan Road No. 209, Xuzhou, 221002, China.
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Neurobiology and Human Anatomy, Xuzhou Medical College, Tongshan Road No. 209, Xuzhou, 221002, China.|Department of Neurosurgery, The First People's Hospital of Lianyungang, Lianyungang, China.
authors.forename: Huizhen
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Neurobiology and Human Anatomy, Xuzhou Medical College, Tongshan Road No. 209, Xuzhou, 221002, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Neurobiology and Human Anatomy, Xuzhou Medical College, Tongshan Road No. 209, Xuzhou, 221002, China.|Department of Neurology, Shuyang People's Hospital, Jiangsu, China.
authors.forename: Fangfang
authors.initials: F
authors.lastname: Chen
authors.affiliation: Department of Neurobiology and Human Anatomy, Xuzhou Medical College, Tongshan Road No. 209, Xuzhou, 221002, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Gao
authors.affiliation: Department of Neurobiology and Human Anatomy, Xuzhou Medical College, Tongshan Road No. 209, Xuzhou, 221002, China.
authors.forename: Shen
authors.initials: S
authors.lastname: Sun
authors.affiliation: Department of Human Anatomy, Histology and Embryology, School of the Basic Medicine, The Fourth Military Medical University, Xi'an, China.
authors.forename: Yunqing
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Neurobiology and Human Anatomy, Xuzhou Medical College, Tongshan Road No. 209, Xuzhou, 221002, China. dshgao@vip.sina.com.|Department of Human Anatomy, Histology and Embryology, School of the Basic Medicine, The Fourth Military Medical University, Xi'an, China. dshgao@vip.sina.com.|Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Jiangsu, China. dshgao@vip.sina.com.
authors.forename: Dianshuai
authors.initials: D
authors.lastname: Gao
mesh_terms: D019002:CD56 Antigen / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000378:metabolism; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000494:pharmacology*; D006801:Humans; D016086:Lentivirus / Q000378:metabolism; D021962:Membrane Microdomains / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D010168:Palmitates / Q000494:pharmacology; D020033:Protein Isoforms / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D019002:CD56 Antigen; D051100:Glial Cell Line-Derived Neurotrophic Factor; C522918:NCAM1 protein, human; D018696:Neuroprotective Agents; D010168:Palmitates; D020033:Protein Isoforms; D034741:RNA, Small Interfering; C022776:2-bromopalmitate; D053148:Caspase 3
keywords: Dopaminergic neuronal cells; Glial cell line-derived neurotrophic factor; Lipid rafts; Neural cell adhesion molecule; Parkinson’s disease