ZFIN ID: ZDB-FIG-181019-35
Yin et al., 2018 - Spatiotemporal Coordination of FGF and Shh Signaling Underlies the Specification of Myoblasts in the Zebrafish Embryo. Developmental Cell   46:735-750.e4 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Conditions:
Anatomical Terms:
Stage Range: 14-19 somites to Prim-5
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage: Prim-5

Fig. 3

FGF Signaling Participates in the Further Differentiation of Both Slow and Fast Muscle Lineages

(A–C) Fluorescent in situ of FGF downstream activators pea3 (A–Aʹʹ) and erm (B–Bʹʹ) and inhibitor spry4 (C–Cʹʹ). Images are taken at the lateral somitic cells (A–C) and the slow muscle fibers (Aʹ–Cʹ) at 18-somite stage. Constructed transverse images are taken at somite S3 and are displayed in (Aʹʹ–Cʹʹ) with lateral to the top and dorsal to the left. Slow muscles are labeled with Prdm1a:GFP and white short arrows. (Cʹʹʹ) Spry4 expression level per cell along the DV axis. The expression of spry4 is measured cell by cell by calculating the average intensity within the nucleus (nCells = 153, from total of 9 somites taken from 5 embryos, NSp > 0.05, Student’s t test).

(D–H) Perturbations of FGF signaling change the number of MPs with MPs identified by Eng2a:eGFP expression. Wild-type embryo (D), SU5402 treatment at 60 μM (E), heat shock of hsp70l:dn-fgfr1-eGFP (F), and heat shock of hsp70l:ca-fgfr1 (G) analyzed, all imaged at 28 hpf at muscle segments 16–20, with quantification of MP number shown in (H), where nSegments = 55, 60, 40, and 40 (from 11, 12, 10, and 10 embryos) for conditions (D–G), respectively.

(I–L) Perturbations of FGF signaling change the number of ECL cells, with ECL cells identified by Pax7 antibody staining. White asterisks label ECL cells with moderate Pax7 intensity. The much brighter cells labeled with white short arrows are neural crest cells. Wild-type embryo (I and Iʹ), SU5402 treatment at 60 μM (J and Jʹ), heat shock of hsp70l:dn-fgfr1 (K and Kʹ) and heat shock of hsp70l:ca-fgfr1 (L and Lʹ) analyzed at 28 hpf at muscle segments 16–20. Transverse views shown in (I–L) with lateral to the top and dorsal to the left.

(M) Quantification of ECL cell number, where nSegments = 16, 21, 15, and 19 (from 6 embryos in each condition) for conditions (I–L), respectively.

∗∗∗p < 0.001, Student’s t test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP i106Tg control 14-19 somites slow muscle cell IHC
Prim-5 slow muscle cell IHC
i106Tg chemical treatment by environment: SU5402 Prim-5 slow muscle cell IHC
i106Tg; pd1Tg heat shock Prim-5 slow muscle cell IHC
i106Tg; pd3Tg heat shock Prim-5 slow muscle cell IHC
i233Tg control Prim-5 muscle pioneer IHC
i233Tg chemical treatment by environment: SU5402 Prim-5 muscle pioneer IHC
i233Tg; pd1Tg heat shock Prim-5 muscle pioneer IHC
i233Tg; pd3Tg heat shock Prim-5 muscle pioneer IHC
etv4 i106Tg control 14-19 somites somite anterior region ISH
etv5b i106Tg control 14-19 somites somite lateral region ISH
spry4 i106Tg control 14-19 somites slow muscle cell ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab-Pax7 IHC i106Tg control Prim-5 skeletal muscle satellite cell
IHC i106Tg chemical treatment by environment: SU5402 Prim-5 skeletal muscle satellite cell
IHC i106Tg; pd1Tg heat shock Prim-5 skeletal muscle satellite cell
IHC i106Tg; pd3Tg heat shock Prim-5 skeletal muscle satellite cell
Phenotype Details
Fish Conditions Stage Phenotype
i106Tg chemical treatment by environment: SU5402 Prim-5 skeletal muscle satellite cell increased amount, abnormal
i106Tg; pd1Tg heat shock Prim-5 skeletal muscle satellite cell increased amount, abnormal
i106Tg; pd3Tg heat shock Prim-5 skeletal muscle satellite cell decreased amount, abnormal
i233Tg chemical treatment by environment: SU5402 Prim-5 muscle pioneer increased amount, abnormal
i233Tg; pd1Tg heat shock Prim-5 muscle pioneer increased amount, abnormal
i233Tg; pd3Tg heat shock Prim-5 muscle pioneer decreased amount, 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, 46, Yin, J., Lee, R., Ono, Y., Ingham, P.W., Saunders, T.E., Spatiotemporal Coordination of FGF and Shh Signaling Underlies the Specification of Myoblasts in the Zebrafish Embryo, 735-750.e4, Copyright (2018) with permission from Elsevier. Full text @ Dev. Cell