Extended Data Fig. 1: Characterisation of the Tg(mbp:TRPV1-tagRFPt) zebrafish model. | Nature Neuroscience

Extended Data Fig. 1: Characterisation of the Tg(mbp:TRPV1-tagRFPt) zebrafish model.

From: New oligodendrocytes exhibit more abundant and accurate myelin regeneration than those that survive demyelination

Extended Data Fig. 1

(a) Schematic illustrating the Tg(mbp:TRPV1-tagRFPt) demyelination model made using Biorender. The rat ortholog of the TRPV1 channel is expressed in myelinating oligodendrocytes and is activated by addition of csn which drives cation influx. Zebrafish TRPV1 channels are insensitive to csn, therefore csn treatment specifically results in damage to myelinating glia which express the rat ortholog of the TRPV1 channel. (b) Confocal images of myelinating oligodendrocytes in the Tg(mbp:EGFP; mbp:TRPV1- tagRFPt) zebrafish line at 4dpf showing oligodendrocytes co-expressing EGFP and tagRFPt in the merged image. Scale bar, 20 µm. (c) Brightfield images of a zebrafish containing the Tg(mbp:TRPV1-tagRFPt) transgene (TRPV1 + ve), or wildtype siblings without the Tg(mbp:TRPV1-tagRFPt) transgene (TRPV1-ve) which show no developmental differences at 4dpf. Scale bars, 500 µm. (d and e) Confocal images of the (d) Tg(mbp:EGFP-CAAX) line and (e) the Tg(mbp:EGFP) line pre-treatment (indicated here as pre-t) at 4dpf, and 3hpt. Zebrafish not containing the Tg(mbp:TRPV1-tagRFPt) transgene show no disruption to myelin or oligodendrocytes following a 2 hour treatment of 10 µM csn. Scale bars, 20 µm. (f) Confocal images of myelin visualised in Tg(mbp:EGFP-CAAX; mbp:TRPV1-tagRFPt) zebrafish, with control (DMSO) and csn treated animals in top and bottom panels respectively pre-treatment (pre-t) at 4dpf, 3hpt, 1dpt and 3dpt. Scale bar, 20 µm.

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