ZFIN ID: ZDB-FIG-090501-27
Peterkin et al., 2009 - Common genetic control of haemangioblast and cardiac development in zebrafish. Development (Cambridge, England)   136(9):1465-1474 Full text @ Development
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: 26+ somites

Fig. 3 Depletion of gata5 and gata6 from the YSL causes cardia bifida but does not affect myelopoiesis. (A) The YSL expression of gata5 and gata6 was depleted by injection of gata5+gata6 morpholinos into the YSL at the 1000-cell stage. To trace the correctly targeted embryos, a fluorescent control morpholino was co-injected. Embryos positive for fluorescence in the YSL alone (+YSL) were selected at several time points and harvested as YSL gata5+gata6-depleted embryos. The embryos showing no fluorescence (-YSL) were collected as negative injection controls and should express wild-type levels of gata5 and gata6. To ensure the efficiency of the morpholinos, positive control embryos were injected with gata5+gata6 morpholinos at the one-cell stage and were fluorescent throughout the embryo (+embryos). To assess heart formation, cmlc2 and nkx2.5 expression was analysed. Depletion of gata5 and gata6 in the YSL (+YSL) showed normal levels of expression of both cmlc2 and nkx2.5, indicating that specification occurs normally in these embryos. However, cardia bifida was observed in the +YSL embryos, demonstrating that gata5 and gata6 are required in the YSL for the correct migration of the cardiac precursors to the midline. By contrast, l-plastin expression was the same as in the wild-type embryos, indicating that gata5 and gata6 expression in the YSL is dispensable for myelopoiesis. Embryos injected at the one-cell stage (+embryos) showed complete absence of cmlc2, nkx2.5 and l-plastin expression, thereby validating morpholino effectiveness. Views are anterior. For each gene and type (┬▒YSL, +embryo) analysed, n=28-38, and the images shown depict the findings for >95% of the embryos. (B,C) Loss of endoderm in casanova morphant embryos but no defects in myelopoiesis. To establish whether endoderm plays a role in myelopoiesis, endodermless embryos were created by injection of casanova morpholinos. Casanova morphants (casmo) were assessed for endoderm formation and myelopoiesis. Myelopoiesis was not affected in cas morphants as l-plastin and mpx remained unaffected (B). Gene expression in the ALM of cas morphants was examined at 5 and 10 somites (C). The formation of myeloid precursors occurred as normal in cas morphants. Expression of pu1 at 5 somites, and runx1, ikaros and cmyb at 10 somites, was unaffected in cas morphants. Views are anterior-dorsal. For each gene analysed n=38-67, and the images shown depict the findings for >95% of the embryos.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
ikzf1 WT control 10-13 somites anterior lateral plate mesoderm ISH
WT + MO1-sox32 standard conditions 10-13 somites anterior lateral plate mesoderm ISH
lcp1 WT control 26+ somites Not Detected endoderm ISH
26+ somites endoderm ISH
Prim-5 common myeloid progenitor ISH
WT + MO1-gata5 + MO1-gata6 standard conditions 26+ somites endoderm ISH
26+ somites Not Detected endoderm ISH
WT + MO1-sox32 standard conditions Prim-5 common myeloid progenitor ISH
mpx WT control Prim-5 common myeloid progenitor ISH
WT + MO1-sox32 standard conditions Prim-5 common myeloid progenitor ISH
myb WT control 10-13 somites anterior lateral plate mesoderm ISH
WT + MO1-sox32 standard conditions 10-13 somites anterior lateral plate mesoderm ISH
myl7 WT control 26+ somites heart rudiment ISH
26+ somites Not Detected heart rudiment ISH
WT + MO1-gata5 + MO1-gata6 standard conditions 26+ somites heart rudiment ISH
26+ somites Not Detected heart rudiment ISH
nkx2.5 WT control 26+ somites Not Detected heart rudiment ISH
26+ somites heart rudiment ISH
WT + MO1-gata5 + MO1-gata6 standard conditions 26+ somites heart rudiment ISH
26+ somites Not Detected heart rudiment ISH
runx1 WT control 10-13 somites anterior lateral plate mesoderm ISH
WT + MO1-sox32 standard conditions 10-13 somites anterior lateral plate mesoderm ISH
spi1b WT control 5-9 somites anterior lateral plate mesoderm ISH
WT + MO1-sox32 standard conditions 5-9 somites anterior lateral plate mesoderm ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-gata5 + MO1-gata6 standard conditions 26+ somites cardioblast migration to the midline involved in heart field formation disrupted, abnormal
26+ somites heart formation disrupted, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development