ZFIN ID: ZDB-FIG-150330-33
Fukui et al., 2014 - S1P-Yap1 Signaling Regulates Endoderm Formation Required for Cardiac Precursor Cell Migration in Zebrafish. Developmental Cell   31:128-136 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 5-9 somites to Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: 5-9 somites to Prim-5

Fig. 4

Endodermal Cells Cell-Autonomously Regulate Their Survival in a Manner Dependent on the Expression of Yap1-Promoted ctgfa

(A) 3D-rendered two-photon laser-scanned z-stack images of Tg(sox17:GFP);(myl7:Nls-mCherry) embryos (28 hpf) injected with MO indicated at the bottom. An asterisk shows the defect of the endoderm. Dorsal view.

(B and C) Incidence of endoderm defects with cardia bifida of Tg(sox17:GFP);(myl7:Nls-mCherry) embryos injected with MO indicated at the bottom. Total number of the embryos analyzed in each morphant group is indicated at the top of the column.

(D and E) Quantitative-PCR analyses of expression of sox17 mRNA (D) and ctgfa mRNA (E) in the endoderm (GFP+ cells) and extra-endoderm (GFP- cells) of the Tg(sox17:GFP) embryos injected with MO indicated at the bottom. GFP+ and GFP- cells from the embryo at 12 hpf were sorted by GFP fluorescence (n = 4).

(F) WISH analyses of ctgfa mRNA expression in the wild-type embryo (left) and in the yap1 morphant (right; 22 hpf). A set of representative images of four independent experiments is shown. Dorsal view.

(G) Schematic illustration of injection of MO into the endoderm-fated cells (simultaneous injection of both acvr1bT206D and sox32 mRNAs) and into the CPC-fated cells (simultaneous injection of both gata5 and smarcd3b mRNAs) with rhodamine dextran to visualize the cells injected with MO and mRNAs. MO, mRNA, and rhodamine dextran were coinjected into one blastomere at the 64-cell stage.

(H) Representative 3D-rendered confocal images of the Tg(sox17:GFP) embryo injected with mRNAs indicated at the top with the MOs indicated above the image as explained in (G).

(I) Quantitative analyses of incidence of endodermal defects in the embryos examined in (H).

(J) Representative 3D-rendered confocal images of the Tg(sox17:GFP);(myl7:Nls-tdEosFP) embryo injected with mRNAs indicated at the top with the MOs indicated above the image as explained in (G).

(K) Quantitative analyses of incidence of heart defects in the embryos examined in (J).

(L and M) Attachment and spread of the endodermal cells obtained at 12 hpf from Tg(sox17:GFP) embryos injected with MO indicated at the top to the fibronectin-coated dish. Arrowheads indicate the cells spreading after attachment to the dish. A panel of representative result of three independent experiments is shown in (L). Scale bars represent 50 µm. (M) The graph shows the number of the spreading GFP-positive cells after attachment. p < 0.01.

See also Figure S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
ccn2a AB standard conditions 26+ somites endoderm ISH
AB + MO1-yap1 standard conditions 26+ somites endoderm ISH
ha01Tg control 5-9 somites endoderm RTPCR
ha01Tg + MO1-yap1 standard conditions 5-9 somites Not Detected endoderm RTPCR
EGFP ha01Tg; ncv11Tg control Prim-5 endoderm IFL
ha01Tg; ncv11Tg + MO1-cyr61 standard conditions Prim-5 endoderm IFL
ha01Tg; ncv11Tg + MO3-ccn2a standard conditions Prim-5 endoderm IFL
mCherry ha01Tg; ncv11Tg control Prim-5 primitive heart tube myocardial precursor IFL
ha01Tg; ncv11Tg + MO1-cyr61 standard conditions Prim-5 primitive heart tube myocardial precursor IFL
ha01Tg; ncv11Tg + MO3-ccn2a standard conditions Prim-5 primitive heart tube myocardial precursor IFL
sox17 ha01Tg control 5-9 somites endoderm RTPCR
ha01Tg + MO1-yap1 standard conditions 5-9 somites endoderm RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-yap1 standard conditions 26+ somites endoderm cellular quality, abnormal
ha01Tg + MO1-yap1 standard conditions 5-9 somites endoderm cellular quality, abnormal
ha01Tg; ncv11Tg + MO1-cyr61 standard conditions Prim-5 cardiac muscle progenitor cell migration to the midline involved in heart field formation occurrence, normal
Prim-5 endoderm anterior region cellular quality, normal
Prim-5 heart morphology, normal
ha01Tg; ncv11Tg + MO1-yap1 + MO3-ccn2a standard conditions Prim-5 cardiac muscle progenitor cell migration to the midline involved in heart field formation decreased occurrence, abnormal
Prim-5 endoderm antero-medial region decreased object quality, abnormal
Prim-5 heart morphogenesis decreased process quality, abnormal
Prim-5 heart split bilaterally, abnormal
Prim-5 myocardial precursor mislocalised laterally, abnormal
ha01Tg; ncv11Tg + MO3-ccn2a standard conditions Prim-5 cardiac muscle progenitor cell migration to the midline involved in heart field formation decreased occurrence, abnormal
Prim-5 endoderm antero-medial region decreased object quality, abnormal
Prim-5 heart morphogenesis decreased process quality, abnormal
Prim-5 heart split bilaterally, abnormal
Prim-5 myocardial precursor mislocalised laterally, 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, 31, Fukui, H., Terai, K., Nakajima, H., Chiba, A., Fukuhara, S., Mochizuki, N., S1P-Yap1 Signaling Regulates Endoderm Formation Required for Cardiac Precursor Cell Migration in Zebrafish, 128-136, Copyright (2014) with permission from Elsevier. Full text @ Dev. Cell