ZFIN ID: ZDB-FIG-191230-1887
Klingseisen et al., 2019 - Oligodendrocyte Neurofascin Independently Regulates Both Myelin Targeting and Sheath Growth in the CNS. Developmental Cell   51(6):730-744.e6 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Pec-fin to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Pec-fin to Protruding-mouth

Figure 6

Neurofascin B Regulates Myelin Sheath Elongation and Stability

(A and A′) Single time-point confocal images of a myelinating process in a Tg(sox10KalTA4, UAS mEGFP) control that makes contact with a target axon (A) and transitions into a recognizable myelin sheath (A′). Scale bar, 5 μm.

(B) Average length of myelin sheaths immediately upon formation in control and nfascb MO-injected animals. Control median = 4.02, 25th percentile 3.45, 75th percentile 4.93, n = 91 from 13 animals and nfascb MO median = 3.67, 25th percentile 3.10, 75th percentile 4.63, n = 195 sheaths from 21 animals, p = 0.0611, Mann–Whitney test.

(C and C′) Confocal images of a myelin sheath elongating over time during the period of sheath formation in a control animal. Arrowheads in all panels denote ends of myelin sheaths. Scale bar, 5 μm.

(D) Average speed of individual sheath elongation in controls over a 12-h period, starting from the hour of their formation during the critical period. Each point represents a single sheath, imaged across 13 animals. No significant difference in speed of growth based on time of sheath formation, p = 0.8933, Kruskal-Wallis test.

(E and E′) Single time-point confocal images of a myelin sheath elongating over time during the critical period of sheath formation in an nfascb MO-injected animal. Scale bar, 5 μm.

(F) Average speed of sheath elongation in nfascb MO-injected animal over a 12-h period, starting from the hour of their formation during the critical period. Each point represents a single sheath, imaged across 21 animals. No significant difference in speed of growth based on time of sheath formation, p = 0.6301, Kruskal–Wallis test.

(G and G′) Single time-point confocal images of a myelin sheath shrinking over time after the critical period in a nfascb MO-injected animal. Scale bar, 5 μm.

(H) Speed of elongation of individual myelin sheaths in control and nfascb MO-injected animals for the first 6 h after their formation and the following 6 h. (control n = 34 sheaths in 8 animals, nfascb MO n = 66 sheaths in 13 animals: 0–6 h: control mean speed 0.77 ± 0.40 SD, nfascb MO mean 0.56 ± 0.41 SD, p = 0.0133, Mann–Whitney-test. 6–12 h: control mean speed 0.89 ± 0.42 SD, nfascb MO mean 0.185 ± 0.60 SD, p < 0.0001, Mann–Whitney test. Note the large number of sheaths shrinking in nfascb MO-injected animals between 6–12 h after sheath formation.

(I and I′) Confocal images of a myelin sheath elongating in a wildtype animal. Scale bar, 10 μm.

(J and J′) Confocal images of a myelin sheath elongating in an nfascbue56 mutant. Scale bar, 10 μm.

(K) Speed of myelin sheath elongation in Tg(mbp:EGFP-CAAX) wildtype and nfascbue56 mutants (control mean speed 0.34 ± 0.40 SD, n = 118 from 6 animals, nfascbue56 mutant mean 0.06 ± 0.23 SD, n = 215 from 9 animals, p < 0.0001, Mann–Whitney test. 16.1% in controls and 36.4% in nfascbue56 represent shrinking myelin sheaths.

(L) Cell body myelination in nfascbue56 mutants. Scale bar, 10 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP nfascbue56/ue56; ue2Tg standard conditions Protruding-mouth oligodendrocyte myelin sheath IFL
Protruding-mouth spinal cord oligodendrocyte IFL
ue2Tg standard conditions Protruding-mouth oligodendrocyte myelin sheath IFL
Protruding-mouth spinal cord oligodendrocyte IFL
GFP ue102Tg; ue7Tg standard conditions Pec-fin oligodendrocyte myelin sheath IFL
Pec-fin spinal cord oligodendrocyte IFL
ue102Tg; ue7Tg + MO1-nfascb standard conditions Pec-fin oligodendrocyte myelin sheath IFL
Pec-fin spinal cord oligodendrocyte IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
nfascbue56/ue56; ue2Tg standard conditions Protruding-mouth spinal cord central nervous system myelin formation decreased rate, abnormal
ue102Tg; ue7Tg + MO1-nfascb standard conditions Pec-fin spinal cord central nervous system myelin formation decreased rate, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 51(6), Klingseisen, A., Ristoiu, A.M., Kegel, L., Sherman, D.L., Rubio-Brotons, M., Almeida, R.G., Koudelka, S., Benito-Kwiecinski, S.K., Poole, R.J., Brophy, P.J., Lyons, D.A., Oligodendrocyte Neurofascin Independently Regulates Both Myelin Targeting and Sheath Growth in the CNS, 730-744.e6, Copyright (2019) with permission from Elsevier. Full text @ Dev. Cell