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Fig. 5. NMJ-capping cells become restricted to the endplate zone postnatally but spread following denervation in adults. (A-C) 2166 immunostaining (green) is diffuse at birth but becomes progressively concentrated near NMJs over the following 1-2 weeks. (D-F) Denervation in adults triggers nestin reactivity and sprouting in terminal Schwann cells within 3 days but no discernible sprouting or nestin immunoreactivity is seen 1 day after axotomy. By contrast, extensive spreading of 2166 immunostaining occurs within 1 day of axotomy (see also supplementary material Fig. S4). At 3 days, some of the Schwann cell sprouts were associated with 2166-positive links between motor endplates (arrowhead) (see also supplementary material Fig. S5). (G) Immunostaining following injection of a BrdU pulse shows BrdU positive staining (red) of nuclei (blue) in both junctional and non-junctional 2166-positive cells (green), suggesting that NMJ-capping cells both divide and spread from endplates following axotomy. Arrowheads indicate BrdU-positive nuclei. Dotted lines indicate the location of three motor endplates, visualized by Alexa-Fluor-647-conjugated -bungarotoxin (not shown). Scale bar: 100 µm (A-C); 50 µm (D-G).

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