Analysis Tools
cellular
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• blastocysts exhibit low reactive oxygen species (ROS) levels, similar to wild-type and heterozygous controls, indicating normal oxidative homeostasis
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• blastocysts show a significantly higher percentage of TUNEL-positive nuclei than controls, indicating increased DNA breaks and cell death
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• blastocysts show a significantly lower percentage of EdU-positive nuclei than controls, indicating decreased cell proliferation
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• blastocysts fail to form typical outgrowth colonies after 3 days in culture
(J:279207)
• in vitro outgrowth assays show that 6 of 11 blastocysts hatch but fail to form a typical inner cell mass (ICM) colony after 3 days in culture
(J:301719)
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embryo
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• blastocysts fail to hatch from the zona pellucida after 3 days in culture
(J:279207)
• in vitro outgrowth assays show that 5 of 11 fail to hatch from the zona pellucida with all blastomeres trapped in the zona after 3 days in culture
(J:301719)
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• embryos fail to develop beyond blastocyst formation
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• both the number of NANOG-positive cells per blastocyst and percentage of NANOG-positive epiblast cells is significantly reduced, indicating a defective epiblast lineage with far fewer cells
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• both the number of SOX17-positive cells per blastocyst and percentage of SOX17-positive primitive endoderm cells is significantly reduced, indicating a defective primitive endoderm lineage with far fewer cells
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• total cell number per blastocyst is significantly lower than in controls
• however, no active TRP53-positive cells are detected, similar to controls
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• number of POU5F1/OCT4-positive cells per blastocyst is severely decreased; percentage of POU5F1/OCT4-positive ICM cells is significantly reduced, indicating far fewer ICM specified cells
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• number of CDX2-positive cells per blastocyst is severely reduced but the percentage of CDX2-positive cells is significantly increased, indicating more trophectoderm (TE) specified cells
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• blastocysts show a significantly lower percentage of EdU-positive nuclei than controls, indicating decreased cell proliferation
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• blastocysts show a significantly higher percentage of TUNEL-positive nuclei than controls, indicating increased DNA breaks and cell death
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• blastocysts fail to form typical outgrowth colonies after 3 days in culture
(J:279207)
• in vitro outgrowth assays show that 6 of 11 blastocysts hatch but fail to form a typical inner cell mass (ICM) colony after 3 days in culture
(J:301719)
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mortality/aging
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IMPC - UCD
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• embryos exhibit peri-implantation lethality; an increased number of empty decidua is noted at E7.5, suggesting that most embryos elicit a decidual response but likely degenerate and resorb with no embryos found at later stages
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• normal looking blastocysts are present in expected Mendelian ratios at E3.5; excess resorption sites but no homozygous embryos are observed at E7.5
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IMPC - UCD
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IMPC - UCD
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reproductive system
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• an increased number of empty decidua is noted at E7.5
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• embryos fail to implant in the uterus
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