title: Integrin-linked kinase regulates oligodendrocyte cytoskeleton, growth cone, and adhesion dynamics.
abstract: Integrin-linked kinase (ILK), a focal adhesion protein, brokers the link between cytoskeleton, cell membrane, and extracellular environment. Here, we demonstrate a role for ILK in laminin-2-mediated adhesion in primary murine oligodendrocytes (OLs) - with ILK loss leading to severe defects in process branching and outgrowth. These defects were partially recovered when the ILK-depleted OLs were instead grown on the non-integrin-activating substrate poly-l-lysine. Intriguingly, ILK loss on the neutral poly-l-lysine substrate led to swelling at the tips of OL processes, which we identified as enlarged growth cones. Employing the bloated ILK-depleted growth cones as template, we demonstrate the appearance of distinct cytoskeletal domains within OL growth cones bearing classic neuronal growth cone architecture. Further, microtubule organization was severely perturbed following ILK loss, with centripetal microtubule looping and failure to bundle occurring in a laminin-2-independent manner. Together, our work highlights differences in specific aspects of OL biology as driven by laminin-2-dependent or independent ILK governed mechanisms. We also reinforce the idea of OLs as growth cone bearing cells and describe the neuronal-like cytoskeleton therein. Finally, we demonstrate a role for ILK in OL growth cone maturation through microtubule regulation, the loss of which translates to decreased process length and myelin production capacity. We describe herein how different substrates fundamentally alter the oligodendrocyte's response to loss of integrin-linked kinase (ILK). On laminin-2 (Ln-2), ILK-depleted oligodendrocytes appear stunted and malformed, while on the non-integrin-activating substrate PLL branching and membrane formation are restored. We also reinforce the idea of oligodendrocytes as growth cone-bearing cells, detailing the growth cone's cytoskeletal architecture. Strikingly, loss of ILK on poly-l-lysine leads to growth cone swelling, the structure's size and motility rendered less dynamic. Together, our work helps reconcile the phenotypic discrepancy between ILK loss in vitro and in vivo, informs on the oligodendrocyte's growth cone, and ascribes a role for ILK in growth cone dynamics.
authors.affiliation: Ottawa Hospital Research Institute, Ottawa, Ontario, Canada.|Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.
authors.forename: John-Paul
authors.initials: JP
authors.lastname: Michalski
authors.affiliation: Ottawa Hospital Research Institute, Ottawa, Ontario, Canada.|Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Cummings
authors.affiliation: Ottawa Hospital Research Institute, Ottawa, Ontario, Canada.|Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.
authors.forename: Ryan W
authors.initials: RW
authors.lastname: O'Meara
authors.affiliation: Ottawa Hospital Research Institute, Ottawa, Ontario, Canada.|Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.|Department of Medicine, University of Ottawa, Ottawa, Ontario, Canada.|University of Ottawa Centre for Neuromuscular Disease, Ottawa, Ontario, Canada.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Kothary
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000166:cytology; D002448:Cell Adhesion / Q000502:physiology*; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism*; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D020439:Growth Cones / Q000502:physiology*; D008239:Lysine / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D018992:Myelin-Associated Glycoprotein / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D000075385:Netrins; D009836:Oligodendroglia / Q000166:cytology*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism; D000099200:Scaffold Protein ILK
chemical_list: D000199:Actins; D049452:Green Fluorescent Proteins; D008239:Lysine; D018992:Myelin-Associated Glycoprotein; D009419:Nerve Tissue Proteins; D000075385:Netrins; D017346:Protein Serine-Threonine Kinases; D014404:Tubulin; D000099200:Scaffold Protein ILK; C456263:Ntng2 protein, mouse; C485184:enhanced green fluorescent protein
keywords: actin; cytoskeleton; growth cone; microtubule; myelin; oligodendrocyte