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Fig. 3. C6st1 mRNA expression is upregulated in migrating Schwann cells and not in Schwann cells that are engaged with axonal fascicles. DRG explants (E14 rats) were cultured on laminin-coated coverslips for 2 days in 10% FBS-DMEM-F12 supplemented with NGF (20 ng/ml). (A) Left panel, phase-contrast view of axons (small arrows) and cells radiating from the explant. Right panel, the same view showing C6st1 mRNA (C6ST) hybridisation signals in cells at the frontier (box); the enlarged inset image of the boxed region reveals the various morphologies of C6st1-expressing cells. (B) In situ hybridisation for C6st1 mRNA combined with SMI31 immunocytochemistry for axons. Left panel, SMI31-positive fibres (small arrows) radiate as networks and fascicles from a DRG explant. C6st1 hybridisation signals were hardly visible in cells associated with axonal fascicles. Towards the frontier (
1090 µm from DRG centre), C6st1-expressing were visible. Right panel, an enlarged view of the frontier zone shows scattered C6st1-expressing cells (arrowheads) apparently in advance of SMI31-positive axons (small arrows). (C) Double immunofluorescence for SMI31-positive axons and S100-positive Schwann cells. S100-positive cells can be seen crowded around the DRG explant (upper right), aligned as chains along SMI31-positive axon fascicles (small arrows), and as scattered cells (arrowheads) in the frontier. The large arrow in each panel indicates the direction of projection of growing axons and emigrating cells from the DRG centre. Bars, 200 µm (A,B), 100 µm (C).
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