Analysis Tools
mortality/aging
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• no homozygous mutant embryos are recovered at E10.5 or later
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embryo
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• TUNEL analysis revealed significant cell death in the allantois at E8.5
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• embryos fail to undergo axial rotation; none of the embryos are turned at E9.5
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• at E8.5 and E9.5, embryos fail to elongate along the anterior-posterior axis and show limited anterior development relative to wild-type controls
• however, embryos undergo normal gastrulation and neuroectodermal, mesodermal and endodermal patterning is unaffected
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• embryos fail to develop beyond the 4-5 somite stage, likely due to extraembryonic defects
• severe growth defects are seen as early as E8.0
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• embryos undergo developmental delay starting around the transition from allantoic bud to head fold stages
• all E8.5 and E9.5 embryos are growth delayed with obvious defects
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• at E8.5 and E9.5, embryos show an overall reduction in size relative to wild-type controls
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• at E8.5, 4 of 27 embryos are still at the late head fold (LHF) stage while the remaining 23 embryos exhibit 0-4 somites, whereas wild-type embryos exhibit 3-11 somites
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• at E9.5, all embryos (17 of 17) exhibit less than 5 somites, whereas all wild-type controls have >13 somites
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• embryos show abnormal development of extraembryonic tissues with defects detected in the amnion, chorion, and visceral yolk sac
• defective extraembryonic development is likely due, at least in part, to reduced BMP2 signaling
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• at E8.5, the allantois is considerably shorter than normal
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• at the early allantoic bud (EB) to late bud (LB) stages, 3 of 24 embryos exhibit clear defects in the amnion and amniochorionic fold (ACF); the proamniotic canal appears wider and the region of the forming amnion is ruffled
• at the early head fold (EHF) to late head fold (LHF) stages, 5 of 7 embryos show ACF defects; ACF is not resolved into amnion and chorion
• at E8.5, all embryos show ACF defects and the posterior amnion is thickened
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• at E8.5, abnormal tissue aggregates composed of chorionic ectoderm are seen above the developing cranial neural folds within the amniotic cavity
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• at the early allantoic bud (EB) to late bud (LB) stages, 3 of 24 embryos exhibit clear defects in the amniochorionic fold (ACF)
• at the early head fold (EHF) to late head fold (LHF) stages, 5 of 7 embryos show ACF defects; ACF is not resolved into amnion and chorion
• at E8.5, a mass of chorionic tissue accumulates at the anterior midline within the amniotic cavity
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• at E8.5, abnormal tissue aggregates composed of chorionic ectoderm are seen above the developing cranial neural folds within the exocoelom
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• embryos exhibit excess or mislocalization of the extraembryonic ectoderm (ExE) component of the amniochorionic fold (ACF) such that the anterior separation point (ASP) contains only ExE; following separation there is ExE derived tissue within the amniotic cavity
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• at E8.5, the visceral yolk sac exhibits abnormal crater-like structures, unlike in wild-type controls
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• at E8.5, the visceral yolk sac lacks the apical vacuoles normally seen in the endoderm; smaller and fewer apical vacuoles are already seen at the late allantoic bud/early head fold stage, suggesting early impairment of the transport functions
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• TUNEL analysis revealed significant cell death in the visceral yolk sac endodermal layer at E8.5
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growth/size/body
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• embryos undergo developmental delay starting around the transition from allantoic bud to head fold stages
• all E8.5 and E9.5 embryos are growth delayed with obvious defects
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• at E8.5 and E9.5, embryos show an overall reduction in size relative to wild-type controls
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nervous system
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• TUNEL analysis revealed significant cell death in the neuroectoderm at E8.5
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• at E8.5, expression of Otx2 (a marker of the anterior neuroectoderm) and of Six3 (normally present in the most rostral neuroectoderm) is either reduced or absent in some embryos
• at E9.0, embryos fail to show robust Sox2 expression in the anterior neuroectoderm, unlike wild-type controls
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cellular
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• TUNEL analysis revealed significant cell death in the allantois at E8.5
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• TUNEL analysis revealed significant cell death in the neuroectoderm at E8.5
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