Myelin inhibits oligodendroglial maturation and regulates oligodendrocytic transcription factor expression
Version of Record online: 10 JUL 2013
Copyright © 2013 Wiley Periodicals, Inc.
Volume 61, Issue 9, pages 1471–1487, September 2013
How to Cite
Plemel, J. R., Manesh, S. B., Sparling, J. S. and Tetzlaff, W. (2013), Myelin inhibits oligodendroglial maturation and regulates oligodendrocytic transcription factor expression. Glia, 61: 1471–1487. doi: 10.1002/glia.22535
- Issue online: 26 JUL 2013
- Version of Record online: 10 JUL 2013
- Manuscript Accepted: 26 APR 2013
- Manuscript Revised: 20 APR 2013
- Manuscript Received: 19 JUN 2012
- Michael Smith Foundation of Health Research
- Canadian Institute for Health Research
- Rick Hansen Man in Motion Foundation
- Multiple Sclerosis Society of Canada
Additional Supporting Information may be found in the online version of this article.
|glia22535-sup-0001-suppFig1.tif||14979K||Supplemental Figure 1: Details of the myelin spot assay. Myelin (1µl) was placed on a coverslip in 3–4 spot roughly equidistant from one another (A). Markers of immature (O4; red) or mature oligodendrocytes, but not OPC markers (NG2; green) diffusely label the myelin spot. These markers were used to determine if the cells were located “On Myelin” or “Off Myelin” (B). Due to the granular appearance of myelin, it was possible to distinguish it from cellular structures. Thus, myelin was not a source of false positives during the quantification of cellular markers. Nuclei are stained with Hoechst (blue).|
|glia22535-sup-0002-suppFig2.tif||13284K||Supplemental Figure 2: Typical purity of oligodendrocyte culture. OPCs were expanded in mitogens for 1 week, before being plated onto coverslips and fed with media lacking mitogens to induce differentiation. After 2 days in vitro, the majority of OPCs differentiated and express the immature oligodendrocyte marker, O4 (red; A). A small proportion of cells are maintained as OPCs as indicated by the marker NG2 (green; B). NG2 expression and O4 expression are typically mutually exclusive (example with white arrow). Those cells expressing both OPC markers (NG2 or PDGFR-α) and O4, were considered to be O4-positive. There was a small percentage of contaminating astrocytes (GFAP; green) (C). Here, 66% of cells label with O4, 23% of cells label with NG2 and 2% of cells label with GFAP, indicating that the vast majority of cells (89%) express oligodendrocyte lineage markers (D). We did not find evidence of neuronal (βIII-tubulin) or microglial (IBA1) contamination. Nuclei are stained with Hoechst (blue). Error bars are standard error of the mean (D).|
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