FIGURE SUMMARY
Title

Midkine-a Regulates the Formation of a Fibrotic Scar During Zebrafish Heart Regeneration

Authors
Grivas, D., González-Rajal, Á., de la Pompa, J.L.
Source
Full text @ Front Cell Dev Biol
FIGURE 1

mdka expression upon injury. (A) qPCR analysis of mdka in regenerating hearts. Brown-Forsythe and Welch ANOVA tests; **P < 0.01, ***P < 0.001; Mean ± SD. (B–D′) ISH of mdka in intact (B,B′), 3 dpci (C,C′), and 7 dpci (D,D′) hearts. V, ventricle; b.a., bulbus arteriosus. Scale bars, 100 μm. (E–F′′) Fluorescent ISH for mdka followed by immunolabeling for GFP and Aldh1a2 in 7 dpci Tg(wt1b:GFP) hearts. Arrowheads indicate mdka-GFP overlap and arrows mdka-Aldh1a2 overlap. Scale bar E, 100 μm; (F–F′′), 10 μm.

EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Condition:
Anatomical Terms:
Stage: Adult
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Adult
FIGURE 2

Generation of mdka-KO zebrafish. (A) The mdka locus and DNA mutations introduced by CRISPR/Cas9. The red line indicates the CRISPER target site, red letters indicate insertions, and red hyphens deletions. (B) Mdka domain organization and predicted protein mutations. Red letters denote novel amino acids, red hyphen deletions. β stands for beta-strands and the hinge domain is shown in green. (C,D) Whole-mount ISH (WM-ISH) for mdka in 2-day post-fertilization (dpf) mdka+/+ and mdkacn105 embryos. Scale bar, 200 μm. (E) qPCR analysis of mdka and mdkb in 2 dpf mdka+/+ and mdkacn105 embryos. t-test; **P < 0.01; Mean ± SD. (F,G) Immunofluorescence staining of Mdka in 7 dpci mdka+/+ and mdkacn105 hearts. Scale bar, 100 μm. (H) Western blot analysis for Mdka in 7 dpci mdka+/+, mdkacn105, and mdka+/cn105 hearts. Tub, a-Tubulin; kDa, kilodalton.

EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage Range: Long-pec to Adult
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Long-pec to Adult
FIGURE 3

Loss of mdka leads to heart regeneration arrest. (A–C) AFOG staining of 90 dpci mdka+/+, mdka+/cn105, and mdkacn105 hearts. Collagen is shown in blue, fibrin in red, and healthy myocardium in brown. Scale bar, 100 μm. (D) Quantification of the scar area normalized to the total ventricle area. nWT = 14, nKO = 15, nHET = 14. (E) Percentage of collagen in the scar area. nWT = nKO = nHET = 15. Ordinary one-way anova; *P < 0.05; **P < 0.01; Mean ± SD.
FIGURE 4

Transcriptional profiling of regenerating mdkacn105 hearts. (A) Total number of differentially expressed genes identified by RNA-seq. Numbers indicate upregulated genes (red) and downregulated genes (blue). (B,C) Circular plots showing representative differentially expressed genes (left semicircle perimeter) and IPA biofunctions and upstream regulators (right semicircle perimeter). Activation z-score scale: green, repression; magenta, activation; white, unchanged. LogFC scale: red, upregulated; blue, downregulated; white, unchanged. (D) qPCR analysis of mdka+/+ and mdkacn105 7 dpci hearts. t-test; *P < 0.05, **P < 0.01, ***P < 0.001; Mean ± SD. (E,F) Immunofluorescence staining of GFP and BrdU in 7 dpci Tg(fli1a:GFP) heart sections. (E′,F′) Higher magnification of the dashed lines in (A,F). Arrowheads indicate proliferating endothelial cells. Scale bar (E,F) 100 μm; (E′,F′) 10 μm. (G) Quantification of BrdU+/GFP+ endothelial cells. nWT = 13, nKO = 15; t-test; **P < 0.01; Mean ± SD.
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 @ Front Cell Dev Biol
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