title: Production and use of lentivirus to selectively transduce primary oligodendrocyte precursor cells for in vitro myelination assays.
abstract: Myelination is a complex process that involves both neurons and the myelin forming glial cells, oligodendrocytes in the central nervous system (CNS) and Schwann cells in the peripheral nervous system (PNS). We use an in vitro myelination assay, an established model for studying CNS myelination in vitro. To do this, oligodendrocyte precursor cells (OPCs) are added to the purified primary rodent dorsal root ganglion (DRG) neurons to form myelinating co-cultures. In order to specifically interrogate the roles that particular proteins expressed by oligodendrocytes exert upon myelination we have developed protocols that selectively transduce OPCs using the lentivirus overexpressing wild type, constitutively active or dominant negative proteins before being seeded onto the DRG neurons. This allows us to specifically interrogate the roles of these oligodendroglial proteins in regulating myelination. The protocols can also be applied in the study of other cell types, thus providing an approach that allows selective manipulation of proteins expressed by a desired cell type, such as oligodendrocytes for the targeted study of signaling and compensation mechanisms. In conclusion, combining the in vitro myelination assay with lentiviral infected OPCs provides a strategic tool for the analysis of molecular mechanisms involved in myelination.
authors: Department of Anatomy and Neuroscience, The University of Melbourne.
authors: Haley M
authors: HM
authors: Peckham
authors: Department of Anatomy and Neuroscience, The University of Melbourne.
authors: Anita H
authors: AH
authors: Ferner
authors: Department of Anatomy and Neuroscience, The University of Melbourne.
authors: Lauren
authors: L
authors: Giuffrida
authors: Department of Anatomy and Neuroscience, The University of Melbourne; The Florey Institute of Neuroscience and Mental Health Research, The University of Melbourne.
authors: Simon S
authors: SS
authors: Murray
authors: Department of Anatomy and Neuroscience, The University of Melbourne; The Florey Institute of Neuroscience and Mental Health Research, The University of Melbourne; xiaoj@unimelb.edu.au.
authors: Junhua
authors: J
authors: Xiao
mesh_terms: D002478:Cells, Cultured; D018920:Coculture Techniques; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism / Q000821:virology; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000502:physiology*; D009186:Myelin Sheath / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000821:virology*; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000821:virology; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism / Q000821:virology*; D015398:Signal Transduction / Q000502:physiology
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