ZFIN ID: ZDB-FIG-180713-5
Almeida et al., 2018 - Myelination of Neuronal Cell Bodies when Myelin Supply Exceeds Axonal Demand. Current biology : CB   28(8):1296-1305.e5 Full text @ Curr. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Anatomical Terms:
Stage: Day 4
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage: Day 4

Fig. 3

Increasing Oligodendrocyte Number Recapitulates Wrapping of Neuronal Cell Bodies

(A) Double myelin and oligodendrocyte reporter (4 dpf), treated with vehicle DMSO, Skp2C25, or GANT61.

(B and C) Oligodendrocyte (e.g., asterisk) number and myelinated cell bodies (e.g., arrowheads) number is increased in the dorsal spinal cord of Skp2C25-treated animals and reduced in GANT61-treated animals, quantified in (B) (p[Skp2C25-DMSO] = 0.0004, n = 20 DMSO and n = 10 Skp2C25; p[C646-DMSO] < 0.0001, n = 22 DMSO and n = 19 C646; p[GANT61-DMSO] < 0.0001, n = 9 DMSO and n = 13 GANT61; t tests; data normalized to control average) and in (C) (p[Skp2C25-DMSO] < 0.0001, n = 20 DMSO and n = 10 Skp2C25; p[C646-DMSO] < 0.0001, n = 22 DMSO and n = 19 C646; p[GANT61-DMSO] = 0.011, n = 9 DMSO and n = 13 GANT61; t tests; data normalized to control average; GANT61 data and its control plotted on right y axis), respectively.

(D) Plotting all normalized data shows that myelinated cell body number positively correlates to oligodendrocyte number (Pearson’s r correlation and p value displayed for each treatment).

(E) TEM of 4 dpf medial-dorsal spinal cord shows increased number of myelinated axons (red asterisks) in Skp2C25-treated animals whereas controls still have many unmyelinated large-caliber axons (green circles).

(F) Total number of large-caliber (>0.5 μm diameter) axons in the medial-dorsal spinal cord is normal in Skp2C25-treated animals (p = 0.852; n = 4 DMSO and n = 5 Skp2C25; t test).

(G) Myelinated axon number is increased with Skp2C25 (p = 0.006; n = 8 DMSO and n = 9 Skp2C25; t test).

(H) Smallest myelinated axons in Skp2C25-treated animals remain above 0.4 μm diameter (p = 0.961 for all bin comparisons; n = 7 DMSO and n = 9 Skp2C25; corrected t tests).

(I) 4 dpf double myelin and Rohon-Beard (RB) neuron reporter (Tg(isl1(ss):Gal4; UAS:DsRed)), focusing in the dorsal spinal cord.

(J and K) Skp2C25-treated animals have more “very dorsal” myelin sheaths (arrows), quantified in (J) (p = 0.011; n = 19 DMSO and n = 19 Skp2C25; t test), which add up to a higher total myelin sheath length, quantified in (K) (p = 0.013; n = 19 DMSO and n = 19 Skp2C25; t test), as well as readily identifiable myelinated cell bodies (arrowheads). Examples of very rare myelin sheaths on dsRed+ large-caliber RB axons (arrows in merged panels), which are not increased with Skp2C25 (see main text for details), are shown.

All graphs display mean and SD. The scale bars represent 20 μm (A) and 10 μm (I). See also Figure S1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP ue1Tg ; ue2Tg control Day 4 oligodendrocyte cytoplasm IFL
Day 4 oligodendrocyte myelin sheath IFL
Day 4 oligodendrocyte plasma membrane IFL
ue1Tg ; ue2Tg chemical treatment by environment: GANT61 Day 4 oligodendrocyte cytoplasm IFL
Day 4 oligodendrocyte myelin sheath IFL
Day 4 oligodendrocyte plasma membrane IFL
ue1Tg ; ue2Tg chemical treatment by environment: SZL-P1-41 Day 4 oligodendrocyte cytoplasm IFL
Day 4 oligodendrocyte myelin sheath IFL
Day 4 oligodendrocyte plasma membrane IFL
zf234Tg ; ue2Tg control Day 4 oligodendrocyte myelin sheath IFL
Day 4 oligodendrocyte plasma membrane IFL
zf234Tg ; ue2Tg chemical treatment by environment: SZL-P1-41 Day 4 oligodendrocyte myelin sheath IFL
Day 4 oligodendrocyte plasma membrane IFL
RFP zf234Tg ; ue2Tg control Day 4 Rohon-Beard neuron axon IFL
Day 4 Rohon-Beard neuron cell body IFL
zf234Tg ; ue2Tg chemical treatment by environment: SZL-P1-41 Day 4 Rohon-Beard neuron axon IFL
Day 4 Rohon-Beard neuron cell body IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ue1Tg ; ue2Tg chemical treatment by environment: C646 Day 4 oligodendrocyte myelin sheath abutting neuron cell body, abnormal
Day 4 spinal cord has extra parts of type oligodendrocyte, abnormal
Day 4 spinal cord spinal cord oligodendrocyte cell differentiation increased occurrence, abnormal
ue1Tg ; ue2Tg chemical treatment by environment: GANT61 Day 4 spinal cord has fewer parts of type oligodendrocyte, abnormal
Day 4 spinal cord spinal cord oligodendrocyte cell differentiation decreased occurrence, abnormal
ue1Tg ; ue2Tg chemical treatment by environment: SZL-P1-41 Day 4 oligodendrocyte myelin sheath abutting neuron cell body, abnormal
Day 4 spinal cord has extra parts of type oligodendrocyte, abnormal
Day 4 spinal cord spinal cord oligodendrocyte cell differentiation increased occurrence, abnormal
zf234Tg ; ue2Tg chemical treatment by environment: SZL-P1-41 Day 4 oligodendrocyte myelin sheath abutting neuron cell body, abnormal
Day 4 Rohon-Beard neuron axon lacks all parts of type oligodendrocyte myelin sheath, normal
Day 4 Rohon-Beard neuron central nervous system myelination arrested, normal
Day 4 spinal cord has extra parts of type oligodendrocyte, abnormal
Day 4 spinal cord spinal cord oligodendrocyte cell differentiation increased occurrence, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Current biology : CB for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Curr. Biol.