Abstract
BackgroundThe decline of remyelination in chronic multiple sclerosis (MS) is in part attributed to inadequate oligodendrocyte precursor cell (OPC) migration, a process governed by the extracellular matrix (ECM). Elucidating the mechanisms underlying OPC migration is therefore an important step towards developing new therapeutic strategies to promote myelin repair. Many seminal OPC culture methods were established using rat-sourced cells, and these often need modification for use with mouse OPCs due to their sensitive nature. It is of interest to develop mouse OPC assays to leverage the abundant transgenic lines. To this end, we developed a new OPC migration method specifically suited for use with mouse-derived cells.ResultsTo validate its utility, we combined the new OPC migration assay with a conditional knockout approach to investigate the role of integrin-linked kinase (ILK) in OPC migration. ILK is a focal adhesion protein that stabilizes cellular adhesions to the extracellular matrix (ECM) by mediating a linkage between matrix-bound integrin receptors and the cytoskeleton. We identified ILK as a regulator of OPC migration on three permissive substrates. ILK loss produced an early, albeit transient, deficit in OPC migration on laminin matrix, while migration on fibronectin and polylysine was heavily reliant on ILK expression.ConclusionsInclusively, our work provides a new tool for studying mouse OPC migration and highlights the role of ILK in its regulation on ECM proteins relevant to MS.
Highlights
The decline of remyelination in chronic multiple sclerosis (MS) is in part attributed to inadequate oligodendrocyte precursor cell (OPC) migration, a process governed by the extracellular matrix (ECM)
By combining the OPCA assay with conditional knockout genetics, we reveal a role for the focal adhesion protein integrin-linked kinase (ILK) in OPC migration on three substrates
The backwashed media was transferred to a Petri dish, and OPCAs were picked with a micropipette by aid of a stereomicroscope
Summary
The decline of remyelination in chronic multiple sclerosis (MS) is in part attributed to inadequate oligodendrocyte precursor cell (OPC) migration, a process governed by the extracellular matrix (ECM). It is of interest to develop mouse OPC assays to leverage the abundant transgenic lines To this end, we developed a new OPC migration method suited for use with mouse-derived cells. Our understanding of OL biology has vastly improved through the study of rat OLs in vitro, for which several isolation methods exist [4,5,6]. These strategies are often not ideal for isolating mouse OPCs for a number of reasons. Researchers have endeavored to modify existing rat OPC protocols/assays to allow for their use with mouse-derived cells [7, 9,10,11] to take advantage of the broad spectrum of transgenic mouse lines
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