Analysis Tools
embryo
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• embryos show a complete lack of nuclear EFTUD2 (a key component of U5 spliceosome) with no reduction of Eftud2 or other U5 or R2TP/PFDL mRNA levels, and a significant increase in intronic transcripts, indicating global splicing defects during early embryonic development
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• in vitro, embryos fail to hatch from the zona pellucida after 24 h of culture in outgrowth media
(J:279207)
• in vitro, embryos fail to hatch from the zona and mostly collapse/die after 24 h of culture in outgrowth media
(J:354661)
• however, embryos appear more expanded after exogenous FGF4 treatment, with 20% initiating hatching in KSOM + heparin + FGF4 culture whereas hatching is never observed without FGF4
(J:354661)
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• at E3.5, embryos show a complete absence of SOX17+ primitive endoderm (PrE) cells
(J:279207)
• embryos show a complete absence of SOX17+ PrE cells in KSOM + heparin culture
(J:354661)
• although some embryos show a few SOX17+ cells in KSOM + heparin + FGF4 culture, exogenous FGF4 treatment does not fully rescue the SOX17 phenotype
(J:354661)
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• embryos show depletion of the critical pluripotency gene SOX2 and PrE marker SOX17
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• embryos exhibit fragmented DNA, as indicated by scattered DAPI signal
(J:279207)
• at E3.5 + 24 h, embryos show a nearly 50% reduction in total cell count along with fragmented DNA/nuclei, as indicated by scattered DAPI signal
(J:354661)
• loss of Sox2 mRNA expression coincides with the collapse of blastocysts
(J:354661)
• total cell count is significantly reduced in KSOM + heparin culture
(J:354661)
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• at E3.5, embryos show a complete absence of SOX2+ inner cell mass (ICM) cells
(J:279207)
• at E3.5 + 24 h, embryos show a complete absence of SOX2 expression, suggesting defective lineage specification in the ICM
(J:354661)
• proportion of ICM cells per total cell number is significantly reduced in KSOM + heparin culture
(J:354661)
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• at E3.5, NANOG (an epiblast marker) is not turned off properly in CDX2+ trophectoderm (TE) cells
(J:279207)
• analysis of tight junctions in expanded blastocysts shows irregular accumulation and breaks in ZO-1 membrane localization between cells, indicating TE cell-cell junction defects
(J:354661)
• NANOG is abnormally expressed in CDX2+ TE cells in KSOM + heparin culture
(J:354661)
• however, exogenous FGF4 treatment largely rescues the aberrant expression of NANOG in TE cells, restricting NANOG expression to the epiblast
(J:354661)
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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, they cannot expand further or hatch out in vitro; many embryos become collapsed or found dead after 24 h culture
(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)
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IMPC - JAX
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reproductive system
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• an increased number of empty decidua (22%) is found at E7.5, suggesting that decidualization may be triggered
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