Localization of internodal and paranodal adhesion molecules Mag, Cadm3, and Caspr in the zebrafish spinal cord.
(A) Confocal image shows myelinating OL expressing the glial Mag fusion protein, labeled using olig1:mag-EGFP. Mag-EGFP (green) co-expressed with myelin reporter, sox10:mRFP (magenta), localizes to the leading edge of myelin, ensheathing the Mauthner axon. Arrowheads indicate the cytoplasm-rich areas of the spirally wrapping myelin sheaths. (B) Confocal image shows a myelinating OL in the dorsal spinal cord expressing glial Mag fusion protein (green) at 3 dpf together with a myelin reporter, sox10:mRFP (magenta). (C) Confocal image shows the accumulation of an axonal protein Cadm3 fusion protein (yellow), labeled using huC:cadm3-EGFP, at myelin internodes. Myelin sheaths are labeled using the reporter sox10:mRFP (magenta). (D) Selected frames from confocal time-lapse recording showing the re-distribution of Caspr-EYFP fusion protein (yellow) to paranodes (left: huC:Gal4;UAS:caspr-EYFP, right: Caspr-EYFP projected on myelin reporter, sox10:mRFP (magenta)). (E) Quantification of the percentage of Caspr-EYFP localization at the paranode of myelin sheaths at 2.5, 3, and 5 dpf. Graph represents mean values ± SD, analyzed by Brown-Forsythe and Welch ANOVA with Dunnett’s T3 multiple comparisons test. *p < 0.05, **p < 0.01, ****p < 0.0001. Scale bar is 10 µm for (A and B) and 5 µm for (C and D). Standard deviation projections are shown for (A–C), and maximum intensity projections are shown for (D). The data underlying this Figure can be found in S1 Data.
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