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tnni3xenopus   

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Experiment details for tnni3

Gessert S et al. (2008) Assay

DM-GRASP/ALCAM/CD166 is required for cardiac morphogenesis and maintenance of cardiac identity in first heart field derived cells.

Gene Clone Species Stages Anatomy
tnni3.L laevis NF stage 24 heart primordium
tnni3.L laevis NF stage 29 and 30 heart primordium

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  Fig. 2. Expression of DM-GRASP in the FHF of Xenopus laevis. (A) Expression of different cardiac markers as determined by whole mount in situ hybridization at different stages as indicated. Arrows indicate cardiac progenitor cell population at stage 20, first heart field (FHF) at stage 24 and cells of the developing primary heart tube (PHT) at stage 29. Arrowheads indicate second heart field (SHF) at stage 24 and 29. The cement gland is highlighted by a dotted line. (B) Schematic representation of the location of cardiac progenitor cells, first heart field (FHF), second heart field (SHF) and forming primary heart tube (PHT) with the corresponding marker genes. DM-GRASP is not expressed in the cardiac progenitor cell population but in the FHF and the forming PHT at later stages. Note that at this stage, the linear heart tube has not yet formed.

Gene Clone Species Stages Anatomy
tnni3.L laevis NF stage 28 heart , cardiac mesoderm , heart primordium

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  Fig. 5. Loss of DM-GRASP affects marker gene expression in the FHF but not the SHF. (A) DM-GRASP MO was unilaterally injected at the 8-cell stage and expression of cardiac marker genes was monitored at stage 20. No changes for Tbx20, Isl-1 and Nkx2.5 could be detected. Anterior views of whole embryos are given. (B) DM-GRASP MO was unilaterally injected at the 8-cell stage and expression of cardiac marker genes as indicated was monitored at stage 28. Loss of DM-GRASP resulted in down-regulation of some marker genes of the FHF (arrows) but not SHF such as Isl-1 or BMP-4. Ventral views of embryos are given. (C) Quantitative presentation of observed phenotype in panel B. Error bars are standard errors of the means. N = number of examined embryos, n = number of independent experiments.

Gene Clone Species Stages Anatomy
tnni3.L laevis NF stage 28 heart , heart primordium

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  Fig. 6. ?5 -UTR DM-GRASP RNA can rescue the loss of marker gene expression in DM-GRASP MO injected embryos. (A) DM-GRASP MO was unilaterally injected at 8-cell stage together with RNA coding for ?5 UTR DM-GRASP or GFP. Expression of cardiac marker genes was monitored at stage 28. (B) Quantitative presentation of observed phenotype. Error bars are standard errors of the means. N = number of examined embryos, n = number of independent experiments.

Gene Clone Species Stages Anatomy
tnni3.L laevis NF stage 28 heart , myocardium , heart primordium

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  Fig. 7. DM-GRASP and Wnt11-R are functionally linked. (A) Wnt11-R MO was unilaterally injected at the 8-cell stage and expression of cardiac marker genes was monitored at stage 28. Wnt11-R does not affect expression of the early cardiac marker genes Nkx2.5, Isl-1, and Tbx20, but markers of differentiated cardiomyocytes of the FHF such as TnIc, cActin, and MHCα. Bar graphs depict the quantitative presentation of the observed phenotype. Error bars are standard errors of the means. N = number of examined embryos, n = number of independent experiments. (B) Co-injection of RNA coding for DM-GRASP partially reverted the phenotype triggered by the Wnt11-R MO. Bar graphs depict the quantitative presentation of observed phenotype. Error bars are standard errors of the means. N = number of examined embryos, n = number of independent experiments.