GATA3 Is Selectively Expressed in the Trophectoderm of Peri-implantation Embryo and Directly Regulates Cdx2 Gene Expression

Abstract

During early mammalian development, genesis of the first two cell lineages, inner cell mass (ICM) and trophectoderm (TE), is dependent upon functions of key transcription factors that are expressed in a regulated and spatially restricted fashion. In this study, we demonstrate that during early mouse development, mRNA expression of transcription factor GATA3 is induced at the 4-cell stage and is consistently present during pre-implantation embryonic development. Interestingly, at the blastocyst stage, Gata3 mRNA is selectively up-regulated within the TE lineage, and GATA3 protein is abundantly present only in the TE but not in the ICM. Using mouse trophoblast stem cells (TS cells) as a model, we found that, knockdown of GATA3 by RNA interference (RNAi) down-regulates expression of caudal-type homeobox 2 (CDX2), a key regulator of the TE lineage. Chromatin immunoprecipitation (ChIP) analyses revealed that, in TS cells, GATA3 directly regulates Cdx2 transcription from a conserved GATA motif at the intron 1 region of the Cdx2 locus. ChIP analyses with mouse blastocysts also detected GATA3 occupancy at intron 1 of the Cdx2 locus. In addition, down-regulation of GATA3 in pre-implantation mouse embryos reduces Cdx2 expression and inhibits morula to blastocyst transformation. Our results indicate a novel function of GATA3, in which it is selectively expressed in TE, regulates expression of key genes in TE lineage, and is involved in morula to blastocyst transformation. During early mammalian development, genesis of the first two cell lineages, inner cell mass (ICM) and trophectoderm (TE), is dependent upon functions of key transcription factors that are expressed in a regulated and spatially restricted fashion. In this study, we demonstrate that during early mouse development, mRNA expression of transcription factor GATA3 is induced at the 4-cell stage and is consistently present during pre-implantation embryonic development. Interestingly, at the blastocyst stage, Gata3 mRNA is selectively up-regulated within the TE lineage, and GATA3 protein is abundantly present only in the TE but not in the ICM. Using mouse trophoblast stem cells (TS cells) as a model, we found that, knockdown of GATA3 by RNA interference (RNAi) down-regulates expression of caudal-type homeobox 2 (CDX2), a key regulator of the TE lineage. Chromatin immunoprecipitation (ChIP) analyses revealed that, in TS cells, GATA3 directly regulates Cdx2 transcription from a conserved GATA motif at the intron 1 region of the Cdx2 locus. ChIP analyses with mouse blastocysts also detected GATA3 occupancy at intron 1 of the Cdx2 locus. In addition, down-regulation of GATA3 in pre-implantation mouse embryos reduces Cdx2 expression and inhibits morula to blastocyst transformation. Our results indicate a novel function of GATA3, in which it is selectively expressed in TE, regulates expression of key genes in TE lineage, and is involved in morula to blastocyst transformation. Genesis of the trophectoderm (TE) 2The abbreviations used are: TEtrophectodermICMinner cell massEGFPenhanced green fluorescent proteinChIPchromatin immunoprecipitation assayEomeseomesoderminshRNAshort-hairpin RNAMEFmouse embryonic fibroblastsRTreverse transcriptaseTSmouse trophoblast stem cellsEShuman embryonic stem cellsUTRuntranslated regionCDX2caudal-type homeobox 2. 2The abbreviations used are: TEtrophectodermICMinner cell massEGFPenhanced green fluorescent proteinChIPchromatin immunoprecipitation assayEomeseomesoderminshRNAshort-hairpin RNAMEFmouse embryonic fibroblastsRTreverse transcriptaseTSmouse trophoblast stem cellsEShuman embryonic stem cellsUTRuntranslated regionCDX2caudal-type homeobox 2. and inner cell mass (ICM) lineages during early mouse development appears to occur in two stages (1Rossant J. Semin. Cell Dev. Biol. 2004; 15: 573-581Crossref PubMed Scopus (96) Google Scholar, 2Zernicka-Goetz M. Nat. Rev. Mol. Cell Biol. 2005; 6: 919-928Crossref PubMed Scopus (85) Google Scholar, 3Kunath T. Strumpf D. Rossant J. Placenta. 2004; 25: S32-S38Crossref PubMed Scopus (72) Google Scholar). First, cells are allocated to different inside and outside positions via asymmetric divisions. Then, the cells in these different positions become specified, and they become committed to restricted developmental fates. Outside cells become committed to the TE, and inside cells become ICM. Development of ICM and TE is regulated by key transcription factors that specify TE and ICM cell fate, and CDX2 has been implicated in this process (4Chawengsaksophak K. James R. Hammond V.E. Köntgen F. Beck F. Nature. 1997; 386: 84-87Crossref PubMed Scopus (554) Google Scholar, 5Strumpf D. Mao C.A. Yamanaka Y. Ralston A. Chawengsaksophak K. Beck F. Rossant J. Development. 2005; 132: 2093-2102Crossref PubMed Scopus (832) Google Scholar, 6Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (902) Google Scholar). Multiple studies indicated the importance of CDX2 in TS cell proliferation, proper function of TE, and successful implantation of blastocyst (5Strumpf D. Mao C.A. Yamanaka Y. Ralston A. Chawengsaksophak K. Beck F. Rossant J. Development. 2005; 132: 2093-2102Crossref PubMed Scopus (832) Google Scholar, 6Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (902) Google Scholar, 7Jedrusik A. Parfitt D.E. Guo G. Skamagki M. Grabarek J.B. Johnson M.H. Robson P. Zernicka-Goetz M. Genes Dev. 2008; 22: 2692-2706Crossref PubMed Scopus (193) Google Scholar, 8Ralston A. Rossant J. Dev. Biol. 2008; 313: 614-629Crossref PubMed Scopus (269) Google Scholar). However, molecular mechanisms that regulate Cdx2 expression in trophoblast cell lineages are poorly understood. Two other transcription factors, eomesodermin (Eomes) and TEA domain family member 4 (TEAD4), are also implicated in TE development. Mutation studies showed that the lack of Eomes also arrests blastocyst development (9Russ A.P. Wattler S. Colledge W.H. Aparicio S.A. Carlton M.B. Pearce J.J. Barton S.C. Surani M.A. Ryan K. Nehls M.C. Wilson V. Evans M.J. Nature. 2000; 404: 95-99Crossref PubMed Scopus (484) Google Scholar). However, Cdx2 is still expressed in Eomes mutants (5Strumpf D. Mao C.A. Yamanaka Y. Ralston A. Chawengsaksophak K. Beck F. Rossant J. Development. 2005; 132: 2093-2102Crossref PubMed Scopus (832) Google Scholar). Tead4 mutants show more severe phenotypes than Cdx2 mutants and are characterized by loss of Cdx2 expression (10Yagi R. Kohn M.J. Karavanova I. Kaneko K.J. Vullhorst D. DePamphilis M.L. Buonanno A. Development. 2007; 134: 3827-3836Crossref PubMed Scopus (374) Google Scholar, 11Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (346) Google Scholar). However, unlike Cdx2, Tead4 expression is not restricted to the TE lineage during pre-implantation development indicating that additional regulatory mechanisms are involved for the restricted expression of Cdx2 in TE lineage. trophectoderm inner cell mass enhanced green fluorescent protein chromatin immunoprecipitation assay eomesodermin short-hairpin RNA mouse embryonic fibroblasts reverse transcriptase mouse trophoblast stem cells human embryonic stem cells untranslated region caudal-type homeobox 2. trophectoderm inner cell mass enhanced green fluorescent protein chromatin immunoprecipitation assay eomesodermin short-hairpin RNA mouse embryonic fibroblasts reverse transcriptase mouse trophoblast stem cells human embryonic stem cells untranslated region caudal-type homeobox 2. Earlier, we found that, among the six members (GATA1–6) of GATA family of transcription factors, only GATA3 is abundantly expressed in TS cells (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar). GATA3 was first cloned as a T cell-specific transcript (13Zheng W. Flavell R.A. Cell. 1997; 89: 587-596Abstract Full Text Full Text PDF PubMed Scopus (1883) Google Scholar, 14Flavell R.A. Li B. Dong C. Lu H.T. Yang D.D. Enslen H. Tournier C. Whitmarsh A. Wysk M. Conze D. Rincon M. Davis R.J. Cold Spring Harb. Symp. Quant. Biol. 1999; 64: 563-571Crossref PubMed Scopus (24) Google Scholar), and germ line deletion of Gata3 results in embryonic lethality due to a multitude of phenotypic abnormalities (15Pandolfi P.P. Roth M.E. Karis A. Leonard M.W. Dzierzak E. Grosveld F.G. Engel J.D. Lindenbaum M.H. Nat. Genet. 1995; 11: 40-44Crossref PubMed Scopus (500) Google Scholar). Interestingly, expression analysis during early mouse development indicated that at embryonic day (E) 8.5, Gata3 expression is only detectable at the ectoplacental cone and trophoblast cells surrounding the embryonic cavity (16George K.M. Leonard M.W. Roth M.E. Lieuw K.H. Kioussis D. Grosveld F. Engel J.D. Development. 1994; 120: 2673-2686Crossref PubMed Google Scholar). Furthermore, it is also shown that in vitro differentiation of human embryonic stem (ES) cells toward trophoblast cell lineage is associated with GATA3 up-regulation (17Xu R.H. Chen X. Li D.S. Li R. Addicks G.C. Glennon C. Zwaka T.P. Thomson J.A. Nat. Biotechnol. 2002; 20: 1261-1264Crossref PubMed Scopus (872) Google Scholar). These results strongly indicate that GATA3 is involved in the transcription factor networks that drive trophoblast cell linage development. Although expression of several trophoblast cell-specific genes is impaired in Gata3 mutant mice (18Ma G.T. Roth M.E. Groskopf J.C. Tsai F.Y. Orkin S.H. 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Nature. 1994; 371: 221-226Crossref PubMed Scopus (1182) Google Scholar, 24Tsai F.Y. Orkin S.H. Blood. 1997; 89: 3636-3643Crossref PubMed Google Scholar), it is predicted that redundant function of another GATA factor, GATA2 might be the cause for this lack of early developmental phenotype in Gata3 mutant mice (18Ma G.T. Roth M.E. Groskopf J.C. Tsai F.Y. Orkin S.H. Grosveld F. Engel J.D. Linzer D.I. Development. 1997; 124: 907-914Crossref PubMed Google Scholar). Our observation that GATA3 negatively regulates Gata2 expression in TS cell population, and loss of GATA3 induces GATA2 expression (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar) in a rat trophoblast stem cell line, further supports this However, trophoblast cell-specific GATA3 expression during early mouse development as as expression in mouse TS cells to that GATA3 might be regulator in TE this in this study, we the expression of GATA3 during early mouse development, and TS cells as a we function the of key genes that are for TE lineage development. we demonstrate that, at the blastocyst stage, GATA3 expression is selectively restricted in the TE lineage. Furthermore, we also demonstrate that GATA3 directly regulates Cdx2 transcription by the intron 1 region of the Cdx2 chromatin Our results indicate that GATA3 is transcription factor for expression in TE lineage during early mammalian development. TS cells a of mouse embryonic fibroblasts in the of factor 4 and in TS cell as (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. 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Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar). and from was from was from and was from and from used as of mouse blastocysts was to the by and D. S. A. PubMed Scopus Google Scholar) with from blastocysts blastocysts for for blastocysts with at for with and with at in for for RNA by the RNA and analysis was as early blastocysts with in and with and in for 1 at with in for at 4 and in in with at at for 1 and and in short-hairpin mouse Gata3 mRNA cloned in in cells as (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar, D. S. Paul S. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar). and of mouse TS cells at without and with TS cells by of for mRNA and protein Gata3 expression of the cells with the Gata3 and which not knockdown GATA3 knockdown GATA3 in pre-implantation GATA3 was cloned in was with that the expression which enhanced green protein the of the was also with and used as in at a of 1 embryos to in both the with of embryos in at in a at cells and a fluorescent and for embryos used for of not to also as day the embryos used to RNA the RNA ChIP analysis with mouse TS cells was as (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar). analysis with for blastocysts with and ChIP analysis was a X. R. 2007; PubMed Scopus (72) Google Scholar) with In chromatin with was used as with and the from and by for GATA3 are in the was used to regulatory of mouse Cdx2 locus. region to of mouse Cdx2 was cloned in region of Cdx2 intron 1 the conserved was cloned of the Cdx2 in the the conserved of the motif was cloned of and of motif was cloned of the conserved GATA motif was and by TS cells with of to of with for at and to the of the was with Cell and was in a the assay for was to the protein of the of Earlier, we found that GATA3 is abundantly expressed in mouse TS cells (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar). GATA3 is selectively expressed in TS cells cells, we a analysis to Gata3 has been that Gata3 be from two distinct and H. M. K.M. J. 2002; PubMed Scopus Google Scholar, O. J. Biol. Chem. 2009; 284: Full Text Full Text PDF PubMed Scopus Google Scholar), two with untranslated first and of these two selectively Gata3 expression in a O. J. Biol. Chem. 2009; 284: Full Text Full Text PDF PubMed Scopus Google Scholar). to these two Gata3 are selectively expressed in TS cells, we found that in TS cells, only are expressed indicating that, in TS cells, the Gata3 In to TS cells, and to mouse embryonic fibroblasts for GATA3 of both and are present in for Gata3 also showed Gata3 mRNA expression in TS cells but not in the and cells not to the analysis also detected GATA3 expression only in TS cells These results indicate that Gata3 is expressed in the stem cells but is in stem the Gata3 mRNA expression at different developmental stages of the pre-implantation mouse we analyses In we detected of Gata3 in and However, Gata3 mRNA is induced from 4-cell and is consistently present to the blastocyst that during pre-implantation development Gata3 is up-regulated at the 4-cell Although Gata3 transcription is induced as early as the 4-cell stage, as GATA3 is expressed selectively in TS cells but not in the we that, at the blastocyst stage, GATA3 expression is restricted in the TE lineage that, we two different In the first we selectively mRNA from the ICM the TE by and Gata3 mRNA expression with to the In the we a to GATA3 protein expression in the both of these we used embryos that are at the early blastocyst stage to the of from the found that, mRNA only from the ICM Gata3 mRNA with that from blastocysts at the blastocyst stage, Gata3 mRNA expression is selectively induced in the TE lineage but not in the ICM lineage. to the of GATA3 protein in the we detected GATA3 protein expression in the TE lineage cells but not in the ICM lineage cells These results that at the blastocyst stage, GATA3 protein expression is restricted at the TE lineage. As GATA3 is selectively expressed in TE lineage cells and TS cells, we GATA3 regulates expression of key genes that are implicated in TE development. we to knockdown GATA3 expression in TS cells and of GATA3 expression of and of to knockdown GATA3 and a that Gata3 mRNA expression by in TS knockdown was also by analysis GATA3 was we the mRNA expression of Cdx2, and found that of GATA3 selectively down-regulates Cdx2 mRNA expression by Eomes mRNA expression not and Tead4 mRNA expression was by of protein expression also the mRNA expression analysis that CDX2 expression was expression was and Eomes expression not with GATA3 These results indicate that, in TS cells, GATA3 regulates expression of a key regulator of the TE lineage. the that GATA3 directly regulates Cdx2 transcription in TS cells, we GATA3 chromatin occupancy at the Cdx2 locus. analysis of 2 to region from the transcription of mouse Cdx2 revealed the of GATA (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar) and only GATA which is at within the intron 1 of the Cdx2 is conserved in mammalian and Two other GATA at and within the region of mouse Cdx2 are conserved mouse and human only conserved GATA motif within intron 1 is within a conserved region Interestingly, analysis in mice from two different revealed that the conserved region of intron 1 is regulatory for Cdx2 expression in J. Mol. Dev. 2007; PubMed Scopus Google Scholar, F. I. Beck F. F. C. E. M. I. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar). we GATA3 the conserved GATA motif within the intron 1 of the Cdx2 locus. In addition, as GATA might also as GATA we GATA3 occupancy at GATA that are within of the mouse Cdx2 locus. However, ChIP analyses in TS cells revealed that, at the Cdx2 GATA3 only the conserved GATA motif of intron 1 but not at other that lack conserved GATA As TS cells are from the TE, and GATA3 is selectively expressed in the TE of the mouse we to GATA3 chromatin occupancy at the Cdx2 intron 1 within Our analysis showed that, to TS cells, GATA3 is at the intron 1 of the Cdx2 in blastocysts a GATA3 region in TS cells (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar) is also by GATA3 in blastocysts However, to TS cells, the GATA motif within the of Cdx2 not show any GATA3 occupancy these results with the ChIP analysis and knockdown studies in TS cells indicate that GATA3 directly regulates Cdx2 expression in TE lineage. the importance of the intron 1 conserved GATA motif in trophoblast cells, we found that to the Cdx2 in of a the conserved intron 1 of the mouse Cdx2 enhanced the by in TS However, of the conserved motif the of the These strongly the motif as for Cdx2 intron 1 Our results show that GATA3 expression is induced as early as 4-cell stage in pre-implantation embryos and is restricted at the trophectoderm in In addition, ChIP analysis that GATA3 directly regulates the Cdx2 in TS cells and However, GATA3 function and of Cdx2 expression might be at than the early stage of trophectoderm development. apparent lack of defect during pre-implantation development in GATA3 mice further supports this However, to the of GATA3 during the early stage of trophectoderm development has not been in a with GATA3 in additional of we to the of GATA3 down-regulation in pre-implantation embryos Cdx2 expression and the pre-implantation development. this we the GATA3 in two cell mouse embryos and pre-implantation development by in that the expression during the of pre-implantation development, we in two cell embryos and expression of at the blastocyst stage also revealed that the of not pre-implantation development of mouse embryos they are in vitro for more to blastocysts and with GATA3 two cell embryos to within 2 and the and the GATA3 showed of However, day of the GATA3 not to blastocysts and additional to the blastocyst stage for additional day not analysis day with embryos that Gata3 mRNA expression was by In addition, analyses also showed that in Cdx2 mRNA expression was by However, loss in Tead4 mRNA was in In addition, mRNA expression of another transcription factor involved in lineage and development of ICM lineage in early embryos H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (902) Google Scholar), was not in embryos These results strongly that GATA3 regulates Cdx2 expression during early trophectoderm development, and GATA3 function is for of blastocysts from the morula networks that and molecular to the and proper function of the TE and ICM lineages are still poorly understood. of the to these molecular mechanisms is to that are selectively expressed in TE ICM lineages by Although studies indicated that GATA3 is expressed in trophoblast cell lineages (12Ray S. Dutta D. Rumi M.A. Kent L.N. Soares M.J. Paul S. J. Biol. Chem. 2009; 284: 4978-4988Abstract Full Text Full Text PDF PubMed Scopus (58) Google Scholar, K.M. Leonard M.W. Roth M.E. Lieuw K.H. Kioussis D. Grosveld F. Engel J.D. Development. 1994; 120: 2673-2686Crossref PubMed Google Scholar, G.T. Roth M.E. Groskopf J.C. Tsai F.Y. Orkin S.H. Grosveld F. Engel J.D. Linzer D.I. Development. 1997; 124: 907-914Crossref PubMed Google Scholar, K.M. Engel J.D. Linzer D.I. Development. 1994; 120: PubMed Google Scholar) and regulates expression of genes in trophoblast cells in (18Ma G.T. Roth M.E. Groskopf J.C. Tsai F.Y. Orkin S.H. Grosveld F. Engel J.D. Linzer D.I. Development. 1997; 124: 907-914Crossref PubMed Google Scholar), have not been to expression during early embryonic development. lack of early developmental phenotype in Gata3 mutant mice might be of the this lack of In this we have the expression of transcription factor GATA3 in pre-implantation embryos that it is selectively induced at the TE lineage at the blastocyst stage, and we have shown that down-regulation of GATA3 inhibits of blastocyst from the morula stage have also that GATA3 directly regulates Cdx2 expression in TS cells and in blastocysts and As CDX2 function is for TE development, we that GATA3 is regulator of molecular that to the early lineage and TE development TS cell In this GATA3 might have a transcription factors and has been shown that during early lineage functions of lineage to to the trophoblast lineage to this in cells W. C. D. W. M. Nat. Cell Biol. 2008; PubMed Scopus Google Scholar). indicated that is for TS cell F. J.A. G. Williams R. J. G. Dev. Biol. 2007; PubMed Scopus Google Scholar). GATA3, both and regulate Cdx2 expression but and not M. A. R. P. Development. 2005; 132: PubMed Scopus Google Scholar, H. M.L. S. Pearce J. R.A. Genes Dev. PubMed Scopus Google Scholar). As the of GATA2 and GATA3 might the lack of early developmental phenotype in mice (18Ma G.T. Roth M.E. Groskopf J.C. Tsai F.Y. Orkin S.H. Grosveld F. Engel J.D. Linzer D.I. Development. 1997; 124: 907-914Crossref PubMed Google Scholar). that GATA2 and GATA3 might function in is from a R. A. R. N. R. M. Ohneda K. Grosveld F. Yamamoto M. S. Blood. 2007; PubMed Scopus Google Scholar), in which both Gata2 and Gata3 expressed in the the of regulatory the defect of However, have not been in a both GATA2 and GATA3 are to during early embryonic development. GATA transcription factors regulate expression in by of a by the of by and by Our that GATA3 directly regulates Cdx2 transcription in a the via which GATA3 regulates Cdx2 expression in TE lineage. is shown that, transcription factor regulates Cdx2 expression during pre-implantation mouse development (10Yagi R. Kohn M.J. Karavanova I. Kaneko K.J. Vullhorst D. DePamphilis M.L. Buonanno A. Development. 2007; 134: 3827-3836Crossref PubMed Scopus (374) Google Scholar, 11Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (346) Google Scholar). analysis in pre-implantation embryos indicated that expression of Cdx2 be detected as early as However, Cdx2 are regulated the of the with in outside cells, and in inside cells A. Rossant J. Dev. Biol. 2008; 313: 614-629Crossref PubMed Scopus (269) Google Scholar). As GATA3 expression is also induced at the outside cells at the blastocyst stage and we that GATA3 induces and of Cdx2 transcription in cells of Cdx2 expression by a other However, other GATA3 and might regulate Cdx2 and other expression be Our also two other are the molecular mechanisms that GATA3 expression during development and expression in the TE lineage at blastocyst and GATA3 of to specify TE and development of trophoblast cell GATA3 it is that might directly regulate GATA3 expression in pre-implantation Our analysis revealed the of conserved within the Gata3 regulatory that expression is induced in TS cells and further a in which GATA3 might negatively regulate to the GATA3 it is to expression GATA3 expression in with this a analysis of GATA3 genes within blastocysts GATA3 function during early embryonic development. J. Soares for with