The interplay between oligodendrocyte (OL) and extracellular matrix (ECM) is critical to the proper maturation of this unique cell type. Recent work has established the β1 integrin-signaling pathway, a mediator for ECM/OL interactions, as an essential component of myelin sheath formation in the central nervous system (CNS). A major downstream effector of β1 integrin is integrin-linked kinase (ILK), an adaptor and structural platform protein. Herein, we (1) generated a model system to study ILK in vivo and (2) employed the model to elucidate ILK’s role in regulating OL biology.
To assess the importance of ILK in OL-mediated myelination, we ablated ILK in primary OLs. ILK loss delayed morphological maturation and led to filamentous actin accumulation in the processes and cell body. Further, we noted an upregulation in RhoA activity, with pathway inhibition rescuing an OL subset. We next moved our studies in vivo. First, we assessed the proteolipid protein promoter’s utility as OL-specific Cre driver. Protocols established, we generated an ILK conditional knockout line (Ilk cKO). Ultrastructural analysis of Ilk cKO optic nerves revealed increased number of amyelinated nerve fibers at P14 with subsequent recovery by P28. The observed transient defects were due neither to a loss nor a gain in total number of mature or progenitor OLs. To rationalize recovery, we grew ILK-depleted OLs on an “inert” substrate. Here, while morphology improved, ILK-depleted OLs were characterized by enlarged and sluggish growth cones as well as microtubule disorganization.
Taken together, our data suggests a role for ILK in regulating the morphological maturation of OLs both in vitro and in vivo, the loss of which results in defective OL branching and membrane formation with phenotype and subsequent recovery dependent upon niche complexity.
| Identifer | oai:union.ndltd.org:uottawa.ca/oai:ruor.uottawa.ca:10393/31611 |
| Date | January 2014 |
| Creators | Michalski, John-Paul |
| Contributors | Kothary, Rashmi |
| Publisher | Université d'Ottawa / University of Ottawa |
| Source Sets | Université d’Ottawa |
| Language | English |
| Detected Language | English |
| Type | Thesis |
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