Analysis Tools
embryo
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• at E5.25, 4 of 20 embryos (presumed to be mutant) are smaller in size
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• at E3.5, embryos exhibit fewer SOX17+ primitive endoderm (PrE) cells than wild-type controls
(J:279207)
• embryos show a 50% reduction of SOX17+ PrE cells in KSOM + heparin culture
(J:354661)
• loss of Fgf4 expression likely results in the reduction of SOX17+ cells and delayed PrE formation
(J:354661)
• however, the percentage of SOX17+ cells is restored to normal levels in KSOM + heparin + FGF4 culture, with no significant change in total cell number after exogenous FGF4 treatment
(J:354661)
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• at E3.5, embryos exhibit fewer SOX2+ inner cell mass (ICM) cells than wild-type controls
(J:279207)
• at E3.5 + 24 h, embryos exhibit only 5% SOX2+ ICM cells versus 12% in control embryos, suggesting defective lineage specification in the ICM; however, embryos show a normal proportion of ICM cells per total cell number in KSOM + heparin culture
(J:354661)
• in vitro outgrowth assays show a much smaller and irregular ICM-derived POU5F1/OCT4+ central colony, suggesting a loss of pluripotency
(J:354661)
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• at E3.5, NANOG (an epiblast marker) is aberrantly expressed in CDX2+ trophectoderm (TE) cells
(J:279207)
• NANOG is abnormally expressed in CDX2+ TE cells in KSOM + heparin culture
(J:354661)
• blastocysts show significantly reduced CDX2 levels in TE cells, in part explaining the aberrant NANOG expression in KSOM culture
(J:354661)
• in vitro outgrowth assays show a more dispersed CDX2high population, and fewer trophoblast giant cell (TGCs) with significantly less cytoplasmic PL-1 (placental lactogen 1) staining, suggesting impaired TE differentiation
(J:354661)
• exogenous FGF4 treatment largely rescues the aberrant expression of NANOG in TE cells, restricting NANOG expression to the epiblast
(J:354661)
• in vivo, E4.0 embryos show normal CDX2 intensity but some CDX2+ TE cells still retain POU5F1/OCT4 expression
(J:354661)
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• in vitro outgrowth assays show fewer TGCs with significantly less cytoplasmic PL-1 staining
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• at E5.25, in 4 of 20 embryos (presumed to be mutant), GATA6+, LAMININ+ parietal endoderm (PE) cells are not tightly adhered to the maternal decidua by TGCs but appear disconnected in uterine crypts
• however, POU5F1/OCT4+ epiblast cells, CDX2+ extraembryonic ectoderm (ExE) cells, and GATA6+ visceral endoderm (VE) cells are consistently observed
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growth/size/body
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• at E5.25, 4 of 20 embryos (presumed to be mutant) are smaller in size
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mortality/aging
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IMPC - JAX
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• although blastocysts are present in expected Mendelian ratios at E3.5, no homozygous embryos can be recovered at E7.5
(J:279207)
• in vitro, embryos can hatch from the zona pellucida and form normal enlarged blastocysts after 24 hours of culture in outgrowth media
(J:279207)
• although morphologically normal blastocysts are found in Mendelian ratios at E3.5, homozygous embryos show post-blastocyst lethality and cannot be recovered at E7.5
(J:354661)
• at E3.5 + 24 h, the total cell count (~80 cells) is comparable to that of control embryos, and no differences in total cell number are seen in KSOM + heparin culture with or without exogenous FGF4
(J:354661)
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IMPC - JAX
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reproductive system
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• an increased number of empty decidua (29%) is found at E7.5, suggesting that decidualization may be triggered
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• implantation defects, observed both in vitro and in vivo, include impaired TE differentiation and defective TGCs
• embryos likely fail to incorporate sufficient H2A.Z (a histone variant of H2A) at the promoter region of Fgf4 and other genes involved in cell projection organization resulting in impaired invasion of trophoblast cells during implantation
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