title: Intracellular Nogo-A facilitates initiation of neurite formation in mouse midbrain neurons in vitro.
abstract: Nogo-A is a transmembrane protein originally discovered in myelin, produced by postnatal CNS oligodendrocytes. Nogo-A induces growth cone collapse and inhibition of axonal growth in the injured adult CNS. In the intact CNS, Nogo-A functions as a negative regulator of growth and plasticity. Nogo-A is also expressed by certain neurons. Neuronal Nogo-A depresses long-term potentiation in the hippocampus and modulates neurite adhesion and fasciculation during development in mice. Here we show that Nogo-A is present in neurons derived from human midbrain (Lund human mesencephalic (LUHMES) cell line), as well as in embryonic and postnatal mouse midbrain (dopaminergic) neurons. In LUHMES cells, Nogo-A was upregulated threefold upon differentiation and neurite extension. Nogo-A was localized intracellularly in differentiated LUHMES cells. Cultured midbrain (dopaminergic) neurons from Nogo-A knock-out mice exhibited decreased numbers of neurites and branches when compared with neurons from wild-type (WT) mice. However, this phenotype was not observed when the cultures from WT mice were treated with an antibody neutralizing plasma membrane Nogo-A. In vivo, neither the regeneration of nigrostriatal tyrosine hydroxylase fibers, nor the survival of nigral dopaminergic neurons after partial 6-hydroxydopamine lesions was affected by Nogo-A deletion. These results indicate that during maturation of cultured midbrain (dopaminergic) neurons, intracellular Nogo-A supports neurite growth initiation and branch formation.
authors.affiliation: Neural Plasticity and Repair Unit, Wallenberg Neuroscience Center, Lund University, BMC A10, 22184 Lund, Sweden; Neurodegeneration and Inflammation Genetics Unit, Wallenberg Neuroscience Center, Lund University, BMC A10, 22184 Lund, Sweden. Electronic address: zuzanna.kurowska@med.lu.se.
authors.forename: Z
authors.initials: Z
authors.lastname: Kurowska
authors.affiliation: Neuronal Survival Unit, Wallenberg Neuroscience Center, Lund University, BMC B11, 221-84 Lund, Sweden; Van Andel Research Institute, Center for Neurodegenerative Science, 333 Bostwick Avenue NE, Grand Rapids, MI 49503, USA.
authors.forename: P
authors.initials: P
authors.lastname: Brundin
authors.affiliation: Brain Research Institute, University of Zürich, Department of Health Sciences and Technology, ETH Zürich, 190 Winterthurerstrasse, 8057 Zürich, Switzerland.
authors.forename: M E
authors.initials: ME
authors.lastname: Schwab
authors.affiliation: Neural Plasticity and Repair Unit, Wallenberg Neuroscience Center, Lund University, BMC A10, 22184 Lund, Sweden.
authors.forename: J-Y
authors.initials: JY
authors.lastname: Li
mesh_terms: D018663:Adrenergic Agents / Q000633:toxicity; D000367:Age Factors; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002452:Cell Count; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D003342:Corpus Striatum / Q000187:drug effects / Q000378:metabolism; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D008636:Mesencephalon / Q000166:cytology*; D051379:Mice; D018345:Mice, Knockout; D009185:Myelin Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016501:Neurites / Q000502:physiology*; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D000070798:Nogo Proteins; D009924:Organ Culture Techniques; D016627:Oxidopamine / Q000633:toxicity
chemical_list: D018663:Adrenergic Agents; D000906:Antibodies; D009185:Myelin Proteins; D000070798:Nogo Proteins; C000603199:RTN4 protein, human; C000603201:Rtn4 protein, mouse; D016627:Oxidopamine
keywords: 3,3′-diaminobenzidine; 4′,6-diamidino-2-phenylindole; 6-OHDA; 6-hydroxydopamine; ANOVA; DAB; DAPI; DMEM; Dulbecco’s Modified Eagle Medium; EDTA; FCS; GDNF; Glial cell line-derived neurotrophic factor; HBSS; HRP; Hanks Balanced Salt Solution; KO; LUHMES cells; Lund human nesencephalic cells; Nogo-A; PBS; PBS with 0.3% Triton X-100; PBST; PFA; Parkinson’s disease; RCF; SDS–PAGE; TH; WT; analysis of variance; cell culture; ethylenediaminetetraacetic acid; fetal calf serum; horseradish peroxidase; knock-out; midbrain dopaminergic neurons; neurite growth; paraformaldehyde; phosphate-buffered saline; relative centrifugal force; sodium dodecyl sulfate polyacrylamide gel electrophoresis; tyrosine hydroxylase; wild-type