ZFIN ID: ZDB-FIG-220628-42
Wiltbank et al., 2022 - Cd59 and inflammation regulate Schwann cell development. eLIFE   11 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Observed In:
Stage: Protruding-mouth

Fig. 3

(A) Schematic of zebrafish cd59 gene. The cd59 coding sequence (357 bp; CDS) encodes the protein domains with the corresponding colors indicated in (C). The non-CDS is indicated in gray. The dashed lines indicate the sgRNA and the target sequence. (B) Genomic sequences for wildtype cd59 and mutant cd59uva48 showing a 15 bp deletion at the splice site between exon 2 (blue) and intron 2 (gray) of the cd59 gene. (C) Schematic of Cd59 protein made in wildtype (119 AA; top panel) and cd59uva48 mutant fish (10–48 AA; bottom panel). (D) Bright-field images of wildtype and cd59uva48 mutant larvae at 72 hours post fertilization (hpf) showing no anatomical defects as a result of cd59 mutation. (E) Chromogenic in situ hybridization (CISH) showing sox10-positive (gray) oligodendrocytes (OLs) in the spinal cord (filled arrows) and Schwann cells (SCs) on the posterior lateral line nerve (pLLN) (empty arrows) at 72 hpf in transverse sections. (F) CISH showing mbp-positive (gray) OLs in the spinal cord (filled arrows) and SCs on the pLLN (empty arrows) at 72 hpf in transverse sections. (G) CISH showing krox20-positive (gray) SCs on the pLLN (empty arrows) and neurons in the brain (filled arrows) at 72 hpf in transverse sections. (H) Fluorescent in situ hybridization (FISH) (RNAscope; ACD) showing cd59 expression (cyan) in sox10:megfp-positive SCs (orange) along the pLLN at 72 hpf. Representative images each display a transverse section (z projection of 20 µm) of single SC on the pLLN. (I) Imaris renderings show cd59 puncta that are localized within each SC. (J) Scatter plot of the number of cd59 RNA puncta in pLLN SCs (mean ± SEM: WT: 64.7 ± 7.6, cd59uva48: 14 ± 1.7; p<0.0001; dot = 1 cell; n = 7 fish per group). These data were compared with Student’s t-test using GraphPad Prism. CISH and FISH images were acquired with confocal imaging. Scale bars: (A) 0.25 mm; (E, F) 25 µm; (G) 50 µm. Artwork created by Ashtyn T. Wiltbank with Illustrator (Adobe).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cd59 cd59uva48/uva48; sl3Tg standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
sl3Tg standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
EGFP cd59uva48/uva48; sl3Tg standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell IHC
sl3Tg standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell IHC
egr2b AB standard conditions Protruding-mouth brain neuron ISH
Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
cd59uva48/uva48 standard conditions Protruding-mouth brain neuron ISH
Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
mbpa AB standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
Protruding-mouth spinal cord oligodendrocyte ISH
cd59uva48/uva48 standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
Protruding-mouth spinal cord oligodendrocyte ISH
sox10 AB standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
Protruding-mouth spinal cord oligodendrocyte ISH
cd59uva48/uva48 standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell ISH
Protruding-mouth spinal cord oligodendrocyte ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
cd59uva48/uva48 standard conditions Protruding-mouth whole organism morphology, normal
cd59uva48/uva48; sl3Tg standard conditions Protruding-mouth posterior lateral line nerve myelinating Schwann cell cd59 expression decreased amount, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Elife