ZFIN ID: ZDB-FIG-191230-1888
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:
Gene:
Fish:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Observed In:
Stage Range: Protruding-mouth to Day 5

Figure 7

Caspr Regulates Myelin Sheath Elongation and Stability, but Not Targeting

(A and B) Confocal images of single oligodendrocytes labeled mosaically using mbp:mCherry-CAAX in wildtype (A) and casprsa12772 mutants at 5 dpf. Scale bar, 10 μm.

(C–E) Number of myelinated cell bodies (C), number of myelin sheaths (D), and the length of myelin sheaths (D), made by individual oligodendrocytes in wildtype, het, and casprsa12772 mutants.

(C) Myelination of cell bodies is increased slightly in casprsa12772 mutant oligodendrocytes (wildtype median 0.0, 25th percentile 0.0, 75th percentile 0.0, n = 15 animals; casprsa12772/+ median 0.0, 25th percentile 0.0, 75th percentile 0.0, n = 35, casprsa12772 median 0.25, 25th percentile 0.0, 75th percentile 1.5 , n = 25 animals; wildtype versus casprsa12772/+ p = 0.8056, wildtype versus casprsa12772 p = 0.0305, casprsa12772/+ versus casprsa12772 p = 0.0156, all Mann–Whitney test.

(D) Number of myelin sheaths per oligodendrocytes is similar in wildtype, casprsa12772/+ and casprsa12772 (wildtype mean 11.78 ± 5.14 SD, n = 15 animals, casprsa12772/+ mean 11.19 ± 3.26 SD, n = 33 animals, casprsa12772 mean 12.00 ± 3.54 SD, n = 24; ANOVA = 0.712).

(E) Myelin sheath length is reduced in casprsa12772 mutants (wildtype mean 29.14 ± 1.71 SEM, n = 15, casprsa12772/+ mean 27.58 ± 1.31 SEM, n = 33 animals, casprsa12772 mean 23.55 ± 1.30 SEM, n = 24 animals; wildtype versus casprsa12772/+ p = 0.4954, wildtype versus casprsa12772 p = 0.0124, casprsa12772/+ versus casprsa12772 p = 0.0374, all t test).

(F and G) Confocal images of the dorsal horn of the spinal cord in wildtype (F) and Caspr mutant (G) mice, indicating the region indicated by dashed lines wherein myelinated cell bodies were searched for. Scale bar, 50 μm.

(F′ and G′) Higher magnification views of regions within the dorsal horn of wild type (F′) and Caspr mutant (G′) animals stained with antibodies that recognize myelin basic protein (green), NeuN (red), and DAPI (blue). Scale bar, 5 μm.

(H) Graph showing that essentially no myelinated cell bodies were observed in wild type or Caspr mutant animals. Wild type mean 0.2800 ± 0.049 SEM, n = 5 (10 sections per mouse), Caspr−⁄− 0.257 ± 0.083 SEM, n = 5 (10 sections per mouse), p = 0.8147, t test.

(I and J) Confocal images of teased fiber preparations taken from wild type (I) and Caspr mutant (J) mice, stained with antibodies that detect Kv1.1 (red), Neurofascin 186 (green), and neurofilament 200 (red). Scale bars, 25 μm.

(K) Quantitation of myelin sheath length in Caspr mutant mice. Caspr internode lengths are reduced compared to wild type; wild type mean 310.7 ± 5.32 SEM, n = 5, Caspr 226.5 ± 8.96 SEM, n = 5 animals, 50 internodes per mouse, p < 0.0001, t test.

(L and L″) Confocal images of a myelin sheath elongating in a wildtype Tg(mbp:EGFP-CAAX) animal. Scale bar, 10 μm.

(M and M′) Confocal images of a myelin sheath elongating in a casprsa12772 mutant. Scale bar, 10 μm.

(N) Speed of myelin sheath elongation in wildtype and casprsa12772 mutants: wildtype mean 0.36 ± 0.35 SD, n = 122 sheaths from 6 animals, casprsa12772 mean 0.16 ± 0.31 SD, n = 151 sheaths from 7 animals, p < 0.0001, Mann–Whitney test. 14.8% in controls and 28.5% in nfascbue56 represent shrinking myelin sheaths.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP cntnap1sa12772/sa12772; 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
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
cntnap1sa12772/+ standard conditions Day 5 spinal cord myelin sheath normal length, normal
Day 5 spinal cord oligodendrocyte has normal numbers of parts of type oligodendrocyte myelin sheath, normal
cntnap1sa12772/sa12772 standard conditions Day 5 spinal cord central nervous system myelin formation process quality, abnormal
Day 5 spinal cord myelin sheath decreased length, abnormal
Day 5 spinal cord myelin sheath surrounding spinal cord neuronal cell body, abnormal
Day 5 spinal cord oligodendrocyte has normal numbers of parts of type oligodendrocyte myelin sheath, normal
cntnap1sa12772/sa12772; ue2Tg standard conditions Protruding-mouth 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