title: N-WASP regulates extension of filopodia and processes by oligodendrocyte progenitors, oligodendrocytes, and Schwann cells-implications for axon ensheathment at myelination.
abstract: The molecular mechanisms used by oligodendrocyte precursor cells (OPCs), oligodendrocytes (OLs), and Schwann cells (SCs) to advance processes for motility in the developing nervous system and to ensheath axons at myelination are currently not well defined. Here we demonstrate that OPCs, OLs, and SCs express the major proteins involved in actin polymerization-driven protrusion; these key proteins including F-actin, the Arp2/3 complex, neural-Wiskott Aldrich Syndrome protein (N-WASP) and WAVE proteins, and the RhoGTPases Rac and Cdc42 are present at the leading edges of processes being extended by OPCs, OLs, and SCs. We reveal by real-time PCR that OLs and SCs have different dominant WAVE isoforms. Inhibition of the WASP/WAVE protein, N-WASP, with wiskostatin that prevents activation of the Arp2/3 complex, blocks process extension by OPCs and SCs. Inhibition of N-WASP also causes OPC and SC process retraction, which is preceded by retraction of filopodia. This implicates filopodia in OPC and SC process stability and also of N-WASP in OPC and SC process dynamics. We also demonstrate that p34 (a component of the Arp2/3 complex), WASP/WAVE proteins, actin, alpha-tubulin, Rac, Cdc42, vinculin, and focal adhesion kinase are detected in water-shocked myelin purified from brain. Inhibition of N-WASP with wiskostatin decreases the number of axons undergoing initial ensheathment in intact optic nerve samples and reduces the Po content of dorsal root ganglia:SC co-cultures. Our findings indicate that OPCs, OLs, and SCs extend processes using actin polymerization-driven protrusion dependent on N-WASP. We hypothesize that inner mesaxons of OLs and SCs use the same mechanism to ensheath axons at myelination.
authors.affiliation: Department of Biology, University of York, York, United Kingdom.
authors.forename: Claire
authors.initials: C
authors.lastname: Bacon
authors.forename: Viktor
authors.initials: V
authors.lastname: Lakics
authors.forename: Laura
authors.initials: L
authors.lastname: Machesky
authors.forename: Martin
authors.initials: M
authors.lastname: Rumsby
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002227:Carbazoles / Q000494:pharmacology; D002460:Cell Line; D018920:Coculture Techniques; D051356:Cortactin / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology; D015870:Gene Expression; D008855:Microscopy, Electron, Scanning; D009186:Myelin Sheath / Q000187:drug effects / Q000502:physiology; D009413:Nerve Fibers, Myelinated / Q000502:physiology*; D009836:Oligodendroglia / Q000502:physiology* / Q000648:ultrastructure; D009900:Optic Nerve / Q000166:cytology; D011412:Propanolamines / Q000494:pharmacology; D011554:Pseudopodia / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D012583:Schwann Cells / Q000502:physiology* / Q000648:ultrastructure; D013234:Stem Cells / Q000502:physiology* / Q000648:ultrastructure; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D002227:Carbazoles; D051356:Cortactin; D011412:Propanolamines; C495770:Wasl protein, rat; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; C489942:wiskostatin