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investigate whether the trophoblast defect was restricted to the production of secondary TGCs (produced by diploid precursors in the ectoplacental cone and derived originally from the polar trophectoderm overlying the inner cell mass of the blastocyst) or also affected the production of primary TGCs (resulting from differentiation of the mural trophectoderm of the blastocyst), blastocysts (3.5 dpc) from crosses between Atrx WT/flox females and GATA1-Cre homozygous transgenic males (Atrx WT/Y;GATA1-Cre +/+) were cultured in vitro for 5 d to monitor outgrowth of the primary trophoblast. After 5 d, individual blastocyst cultures were scored for the extent of primary trophoblast outgrowth, and the Atrx genotype and sex of the blastocyst were determined by PCR. Most blastocysts hatched from the zona pellucida within 24 h, and trophoblast cells spreading out from the inner cell mass could usually be detected within 48 h of culture. No difference was observed in the rate or extent of trophoblast outgrowth over 5 d of culture between Atrxnull blastocysts (Atrx null/Y, n = 6) and blastocysts bearing an Atrx WT allele (Atrx WT/WT, n = 6; Atrx WT/null, n = 6; Atrx WT/Y, n = 6) (examples shown in Figure 7D), suggesting that the defect specifically involves the secondary giant cell compartment. This is consistent with the observation that Atrxnull conceptuses implant successfully and survive to gastrulation. Taken together, these data suggest that loss of Atrx results in a defect in formation of the secondary trophoblast that is apparent from 7.5 dpc. Despite initiating normal organisation in the embryo proper, Atrxnull conceptuses exhibit a proliferative defect by 7.5 dpc and die by around 9.5 dpc, probably due to a nutritional deficit resulting from failure to develop a normal trophoblast."}

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investigate whether the trophoblast defect was restricted to the production of secondary TGCs (produced by diploid precursors in the ectoplacental cone and derived originally from the polar trophectoderm overlying the inner cell mass of the blastocyst) or also affected the production of primary TGCs (resulting from differentiation of the mural trophectoderm of the blastocyst), blastocysts (3.5 dpc) from crosses between Atrx WT/flox females and GATA1-Cre homozygous transgenic males (Atrx WT/Y;GATA1-Cre +/+) were cultured in vitro for 5 d to monitor outgrowth of the primary trophoblast. After 5 d, individual blastocyst cultures were scored for the extent of primary trophoblast outgrowth, and the Atrx genotype and sex of the blastocyst were determined by PCR. Most blastocysts hatched from the zona pellucida within 24 h, and trophoblast cells spreading out from the inner cell mass could usually be detected within 48 h of culture. No difference was observed in the rate or extent of trophoblast outgrowth over 5 d of culture between Atrxnull blastocysts (Atrx null/Y, n = 6) and blastocysts bearing an Atrx WT allele (Atrx WT/WT, n = 6; Atrx WT/null, n = 6; Atrx WT/Y, n = 6) (examples shown in Figure 7D), suggesting that the defect specifically involves the secondary giant cell compartment. This is consistent with the observation that Atrxnull conceptuses implant successfully and survive to gastrulation. Taken together, these data suggest that loss of Atrx results in a defect in formation of the secondary trophoblast that is apparent from 7.5 dpc. Despite initiating normal organisation in the embryo proper, Atrxnull conceptuses exhibit a proliferative defect by 7.5 dpc and die by around 9.5 dpc, probably due to a nutritional deficit resulting from failure to develop a normal trophoblast."}

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After 5 d, individual blastocyst cultures were scored for the extent of primary trophoblast outgrowth, and the Atrx genotype and sex of the blastocyst were determined by PCR. Most blastocysts hatched from the zona pellucida within 24 h, and trophoblast cells spreading out from the inner cell mass could usually be detected within 48 h of culture. No difference was observed in the rate or extent of trophoblast outgrowth over 5 d of culture between Atrxnull blastocysts (Atrx null/Y, n = 6) and blastocysts bearing an Atrx WT allele (Atrx WT/WT, n = 6; Atrx WT/null, n = 6; Atrx WT/Y, n = 6) (examples shown in Figure 7D), suggesting that the defect specifically involves the secondary giant cell compartment. This is consistent with the observation that Atrxnull conceptuses implant successfully and survive to gastrulation. Taken together, these data suggest that loss of Atrx results in a defect in formation of the secondary trophoblast that is apparent from 7.5 dpc. Despite initiating normal organisation in the embryo proper, Atrxnull conceptuses exhibit a proliferative defect by 7.5 dpc and die by around 9.5 dpc, probably due to a nutritional deficit resulting from failure to develop a normal trophoblast."}