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Atsuo Ogura - One of the best experts on this subject based on the ideXlab platform.
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somatic donor cell type correlates with embryonic but not extra embryonic gene expression in postimplantation cloned embryos
PLOS ONE, 2013Co-Authors: Ryutaro Hirasawa, Kimiko Inoue, Atsuo Ogura, Shogo MatobaAbstract:The great majority of embryos generated by somatic cell nuclear transfer (SCNT) display defined abnormal phenotypes after implantation, such as an increased likelihood of death and abnormal placentation. To gain better insight into the underlying mechanisms, we analyzed genome-wide gene expression profiles of day 6.5 postimplantation mouse embryos cloned from three different cell types (cumulus cells, neonatal Sertoli cells and fibroblasts). The embryos retrieved from the uteri were separated into embryonic (epiblast) and Extraembryonic (Extraembryonic Ectoderm and ectoplacental cone) tissues and were subjected to gene microarray analysis. Genotype- and sex-matched embryos produced by in vitro fertilization were used as controls. Principal component analysis revealed that whereas the gene expression patterns in the embryonic tissues varied according to the donor cell type, those in Extraembryonic tissues were relatively consistent across all groups. Within each group, the embryonic tissues had more differentially expressed genes (DEGs) (>2-fold vs. controls) than did the Extraembryonic tissues (P<1.0×10–26). In the embryonic tissues, one of the common abnormalities was upregulation of Dlk1, a paternally imprinted gene. This might be a potential cause of the occasional placenta-only conceptuses seen in SCNT-generated mouse embryos (1–5% per embryos transferred in our laboratory), because dysregulation of the same gene is known to cause developmental failure of embryos derived from induced pluripotent stem cells. There were also some DEGs in the Extraembryonic tissues, which might explain the poor development of SCNT-derived placentas at early stages. These findings suggest that SCNT affects the embryonic and Extraembryonic development differentially and might cause further deterioration in the embryonic lineage in a donor cell-specific manner. This could explain donor cell-dependent variations in cloning efficiency using SCNT.
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Preparation and validation of the samples.
2013Co-Authors: Ryutaro Hirasawa, Kimiko Inoue, Shogo Matoba, Atsuo OguraAbstract:(A) Embryos were retrieved at E6.5 (left) and dissected into four parts (right). EPC, ectoplacental cone; VE, visceral endoderm; EXE, Extraembryonic Ectoderm; EPI, epiblast. Scale bar = 100 µm. B) Raw signal values of Pou5f1 (left), Cdx2 (center) and Pgk1 (right) genes extracted from microarray data. Pou5f1 and Cdx2 were detected exclusively in the embryonic and Extraembryonic samples, respectively, while Pgk1 was detected in both samples. These results confirmed the accuracy of sample preparation from the embryonic and Extraembryonic tissues. Em, embryonic samples; Ex, Extraembryonic samples; IVF, samples from in vitro fertilized control embryos; CC, cumulus cell-derived clone; FC, fibroblast-derived clone; SC, Sertoli cell-derived clone. See Figure S2 for further validation by several other marker genes.
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Somatic Donor Cell Type Correlates with Embryonic, but Not Extra-Embryonic, Gene Expression in Postimplantation Cloned Embryos
2013Co-Authors: Ryutaro Hirasawa, Kimiko Inoue, Shogo Matoba, Atsuo OguraAbstract:The great majority of embryos generated by somatic cell nuclear transfer (SCNT) display defined abnormal phenotypes after implantation, such as an increased likelihood of death and abnormal placentation. To gain better insight into the underlying mechanisms, we analyzed genome-wide gene expression profiles of day 6.5 postimplantation mouse embryos cloned from three different cell types (cumulus cells, neonatal Sertoli cells and fibroblasts). The embryos retrieved from the uteri were separated into embryonic (epiblast) and Extraembryonic (Extraembryonic Ectoderm and ectoplacental cone) tissues and were subjected to gene microarray analysis. Genotype- and sex-matched embryos produced by in vitro fertilization were used as controls. Principal component analysis revealed that whereas the gene expression patterns in the embryonic tissues varied according to the donor cell type, those in Extraembryonic tissues were relatively consistent across all groups. Within each group, the embryonic tissues had more differentially expressed genes (DEGs) (>2-fold vs. controls) than did the Extraembryonic tissues (P
Azim M Surani - One of the best experts on this subject based on the ideXlab platform.
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-5
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.ical removal the VE (-VE) or the ExE (-ExE). (B) E5.75 embryo showing GFP signal in the VE (white arrows). PGC precursors emerge approximately 12 hours later in the proximal epiblast region adjacent to the asterisks. (C, D) Explants derived from E5.75 -VE embryos cultured for 1 day, D1 (C) and 3 days, D3 (D). (E-H) Explants derived from E5.75 whole and -ExE embryos cultured for 1 day, D1 (E, G) and 3 days, D3 (F, H). E, G, white arrows point to Blimp1(GFP)-positive VE; F, white asterisk marks Blimp1(GFP)-positive PGC precursors in the proximal-posterior part of explant; H, red arrow points to Blimp1(GFP)-negative VE, but Blimp1(GFP)-positive epiblast. Cartoons adjacent to the explants elucidate the morphology of the explant. Abbreviations: Emb, tissue derived from epiblast; ExE, Extraembryonic Ectoderm; thick black line, endoderm. (I, J) Explants derived from E5.75 whole and -ExE (I) and (J) embryos cultured for 3 days, D3. White asterisk marks the proximal-posterior part of explant; -ExE explants were inserted in the lower left corner of I and J. K, After 4 days of culture, the cluster of Blimp1(GFP)-positive cells present in explants from whole embryos turns positive for alkaline phosphatase-activity (AP). Scale bar in B-J is 100 μm and K is 25 μm
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-0
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.ical removal the VE (-VE) or the ExE (-ExE). (B) E5.75 embryo showing GFP signal in the VE (white arrows). PGC precursors emerge approximately 12 hours later in the proximal epiblast region adjacent to the asterisks. (C, D) Explants derived from E5.75 -VE embryos cultured for 1 day, D1 (C) and 3 days, D3 (D). (E-H) Explants derived from E5.75 whole and -ExE embryos cultured for 1 day, D1 (E, G) and 3 days, D3 (F, H). E, G, white arrows point to Blimp1(GFP)-positive VE; F, white asterisk marks Blimp1(GFP)-positive PGC precursors in the proximal-posterior part of explant; H, red arrow points to Blimp1(GFP)-negative VE, but Blimp1(GFP)-positive epiblast. Cartoons adjacent to the explants elucidate the morphology of the explant. Abbreviations: Emb, tissue derived from epiblast; ExE, Extraembryonic Ectoderm; thick black line, endoderm. (I, J) Explants derived from E5.75 whole and -ExE (I) and (J) embryos cultured for 3 days, D3. White asterisk marks the proximal-posterior part of explant; -ExE explants were inserted in the lower left corner of I and J. K, After 4 days of culture, the cluster of Blimp1(GFP)-positive cells present in explants from whole embryos turns positive for alkaline phosphatase-activity (AP). Scale bar in B-J is 100 μm and K is 25 μm
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-4
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.aining Blimp1(GFP)-positive PGC precursors isolated and transplanted either distally of proximal-posteriorly into a E6.5 wild type (WT) embryo. The recombinates were cultured for 3 days and stained for LacZ and alkaline phosphatase-activity. (B, C) Sagittal section showing a recombinate explant in which the PGC precursor cluster were placed distally (B) and proximal-posteriorly (C). The inserts are a magnification of the region containing PGCs depicted in box. For orientation, a cartoon of the whole explant is included showing in the dashed box the region of the explant imaged. Scale bar is 200 μm
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-7
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.on of a E5.75 embryo showing the anterior-posterior (A-P) and left-right (L-R) axes. (C) E5.75 embryo cut longitudinally through the L-R axis (red-dashed line in B) showing GFP expression in the AVE. (D) E5.75 embryo cut longitudinally through the L-R axis after 4 days culture stained for alkaline phosphatase-activity. Black arrow in D points to PGCs. Scale bar is 100 μm. (E) Quantification of the number or explant-pairs cut longitudinally through the L-R axis that exhibited PGCs after 4 days of culture in one explant-half (posterior), both explants-halves or in none of the explant-halves
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-2
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.on of a E5.75 embryo showing the anterior-posterior (A-P) and left-right (L-R) axes. (C) E5.75 embryo cut longitudinally through the L-R axis (red-dashed line in B) showing GFP expression in the AVE. (D) E5.75 embryo cut longitudinally through the L-R axis after 4 days culture stained for alkaline phosphatase-activity. Black arrow in D points to PGCs. Scale bar is 100 μm. (E) Quantification of the number or explant-pairs cut longitudinally through the L-R axis that exhibited PGCs after 4 days of culture in one explant-half (posterior), both explants-halves or in none of the explant-halves
Katsuhiko Hayashi - One of the best experts on this subject based on the ideXlab platform.
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-5
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.ical removal the VE (-VE) or the ExE (-ExE). (B) E5.75 embryo showing GFP signal in the VE (white arrows). PGC precursors emerge approximately 12 hours later in the proximal epiblast region adjacent to the asterisks. (C, D) Explants derived from E5.75 -VE embryos cultured for 1 day, D1 (C) and 3 days, D3 (D). (E-H) Explants derived from E5.75 whole and -ExE embryos cultured for 1 day, D1 (E, G) and 3 days, D3 (F, H). E, G, white arrows point to Blimp1(GFP)-positive VE; F, white asterisk marks Blimp1(GFP)-positive PGC precursors in the proximal-posterior part of explant; H, red arrow points to Blimp1(GFP)-negative VE, but Blimp1(GFP)-positive epiblast. Cartoons adjacent to the explants elucidate the morphology of the explant. Abbreviations: Emb, tissue derived from epiblast; ExE, Extraembryonic Ectoderm; thick black line, endoderm. (I, J) Explants derived from E5.75 whole and -ExE (I) and (J) embryos cultured for 3 days, D3. White asterisk marks the proximal-posterior part of explant; -ExE explants were inserted in the lower left corner of I and J. K, After 4 days of culture, the cluster of Blimp1(GFP)-positive cells present in explants from whole embryos turns positive for alkaline phosphatase-activity (AP). Scale bar in B-J is 100 μm and K is 25 μm
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-0
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.ical removal the VE (-VE) or the ExE (-ExE). (B) E5.75 embryo showing GFP signal in the VE (white arrows). PGC precursors emerge approximately 12 hours later in the proximal epiblast region adjacent to the asterisks. (C, D) Explants derived from E5.75 -VE embryos cultured for 1 day, D1 (C) and 3 days, D3 (D). (E-H) Explants derived from E5.75 whole and -ExE embryos cultured for 1 day, D1 (E, G) and 3 days, D3 (F, H). E, G, white arrows point to Blimp1(GFP)-positive VE; F, white asterisk marks Blimp1(GFP)-positive PGC precursors in the proximal-posterior part of explant; H, red arrow points to Blimp1(GFP)-negative VE, but Blimp1(GFP)-positive epiblast. Cartoons adjacent to the explants elucidate the morphology of the explant. Abbreviations: Emb, tissue derived from epiblast; ExE, Extraembryonic Ectoderm; thick black line, endoderm. (I, J) Explants derived from E5.75 whole and -ExE (I) and (J) embryos cultured for 3 days, D3. White asterisk marks the proximal-posterior part of explant; -ExE explants were inserted in the lower left corner of I and J. K, After 4 days of culture, the cluster of Blimp1(GFP)-positive cells present in explants from whole embryos turns positive for alkaline phosphatase-activity (AP). Scale bar in B-J is 100 μm and K is 25 μm
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-4
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.aining Blimp1(GFP)-positive PGC precursors isolated and transplanted either distally of proximal-posteriorly into a E6.5 wild type (WT) embryo. The recombinates were cultured for 3 days and stained for LacZ and alkaline phosphatase-activity. (B, C) Sagittal section showing a recombinate explant in which the PGC precursor cluster were placed distally (B) and proximal-posteriorly (C). The inserts are a magnification of the region containing PGCs depicted in box. For orientation, a cartoon of the whole explant is included showing in the dashed box the region of the explant imaged. Scale bar is 200 μm
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-7
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.on of a E5.75 embryo showing the anterior-posterior (A-P) and left-right (L-R) axes. (C) E5.75 embryo cut longitudinally through the L-R axis (red-dashed line in B) showing GFP expression in the AVE. (D) E5.75 embryo cut longitudinally through the L-R axis after 4 days culture stained for alkaline phosphatase-activity. Black arrow in D points to PGCs. Scale bar is 100 μm. (E) Quantification of the number or explant-pairs cut longitudinally through the L-R axis that exhibited PGCs after 4 days of culture in one explant-half (posterior), both explants-halves or in none of the explant-halves
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Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors-2
2011Co-Authors: Susana Chuva M De Sousa Lopes, Katsuhiko Hayashi, Azim M SuraniAbstract:Copyright information:Taken from "Proximal visceral endoderm and Extraembryonic Ectoderm regulate the formation of primordial germ cell precursors"http://www.biomedcentral.com/1471-213X/7/140BMC Developmental Biology 2007;7():140-140.Published online 20 Dec 2007PMCID:PMC2231376.on of a E5.75 embryo showing the anterior-posterior (A-P) and left-right (L-R) axes. (C) E5.75 embryo cut longitudinally through the L-R axis (red-dashed line in B) showing GFP expression in the AVE. (D) E5.75 embryo cut longitudinally through the L-R axis after 4 days culture stained for alkaline phosphatase-activity. Black arrow in D points to PGCs. Scale bar is 100 μm. (E) Quantification of the number or explant-pairs cut longitudinally through the L-R axis that exhibited PGCs after 4 days of culture in one explant-half (posterior), both explants-halves or in none of the explant-halves
Elizabeth J. Robertson - One of the best experts on this subject based on the ideXlab platform.
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the nodal precursor acting via activin receptors induces mesoderm by maintaining a source of its convertases and bmp4
Developmental Cell, 2006Co-Authors: Nadav Benhaim, Elizabeth J. Robertson, Marcela Guzmanayala, L Pescatore, Daniel Mesnard, Mirko Bischofberger, Felix Naef, Daniel B ConstamAbstract:During early mouse development, the subtilisin-like proprotein convertases (SPC) Furin and PACE4 pattern the primitive Ectoderm and visceral endoderm, presumably by activating the TGFss-related Nodal precursor. Here, mutation of the SPC motif provides direct evidence that Nodal processing is essential to specify anterior visceral endoderm and mesendoderm. Surprisingly, however, the Nodal precursor binds and activates activin receptors to maintain expression of Furin, PACE4, and Bmp4 in Extraembryonic Ectoderm at a distance from the Nodal source. In return, Bmp4 induces Wnt3, which amplifies Nodal expression in the epiblast and mediates induction of mesoderm. We conclude that uncleaved Nodal sustains the Extraembryonic source of proprotein convertases and Bmp4 to amplify Nodal signaling in two nonredundant feedback loops with dual timescales and to localize primitive streak formation at the posterior pole. Based on mathematical modeling, we discuss how these sequential loops control cell fate.
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Control of early anterior-posterior patterning in the mouse embryo by TGF-beta signalling
Philosophical transactions of the Royal Society of London. Series B Biological sciences, 2003Co-Authors: Elizabeth J. Robertson, Dominic P. Norris, Jane Brennan, Elizabeth K. BikoffAbstract:Prior to gastrulation the mouse embryo exists as a symmetrical cylinder consisting of three tissue layers. Positioning of the future anterior-posterior axis of the embryo occurs through coordinated cell movements that rotate a pre-existing proximal-distal (P-D) axis. Overt axis formation becomes evident when a discrete population of proximal epiblast cells become induced to form mesoderm, initiating primitive streak formation and marking the posterior side of the embryo. Over the next 12-24 h the primitive streak gradually elongates along the posterior side of the epiblast to reach the distal tip. The most anterior streak cells comprise the 'organizer' region and include the precursors of the so-called 'axial mesendoderm', namely the anterior definitive endoderm and prechordal plate mesoderm, as well as those cells that give rise to the morphologically patent node. Signalling pathways controlled by the transforming growth factor-beta ligand nodal are involved in orchestrating the process of axis formation. Embryos lacking nodal activity arrest development before gastrulation, reflecting an essential role for nodal in establishing P-D polarity by generating and maintaining the molecular pattern within the epiblast, Extraembryonic Ectoderm and the visceral endoderm. Using a genetic strategy to manipulate temporal and spatial domains of nodal expression reveals that the nodal pathway is also instrumental in controlling both the morphogenetic movements required for orientation of the final axis and for specification of the axial mesendoderm progenitors.
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from fertilization to gastrulation axis formation in the mouse embryo
Current Opinion in Genetics & Development, 2001Co-Authors: Cindy C Lu, Jane Brennan, Elizabeth J. RobertsonAbstract:Abstract Although much remains unknown about how the embryonic axis is laid down in the mouse, it is now clear that reciprocal interactions between the Extraembryonic and embryonic lineages establish and reinforce patterning of the embryo. Atearly post-implantation stages, the Extraembryonic Ectoderm appears to impart proximal–posterior identity to the adjacent proximal epiblast, whereas the distal visceral endoderm signals to the underlying epiblast to restrict posterior identity as it moves anteriorward. At gastrulation, the visceral endoderm is necessary for specifying anterior primitive streak derivatives, which, in turn, pattern the anterior epiblast. Polarity of these Extraembryonic tissues can be traced back to the blastocyst stage, where asymmetry has been linked to the point of sperm entry at fertilization.
Janet Rossant - One of the best experts on this subject based on the ideXlab platform.
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esrrb function is required for proper primordial germ cell development in presomite stage mouse embryos
Developmental Biology, 2019Co-Authors: Eiichi Okamura, Oliver H Tam, Eszter Posfai, Katie Cockburn, Cheryl Q E Lee, Jodi Garner, Janet RossantAbstract:Estrogen related receptor beta (Esrrb) is an orphan nuclear receptor that is required for self-renewal and pluripotency in mouse embryonic stem (ES) cells. However, in the early post-implantation mouse embryo, Esrrb is specifically expressed in the Extraembryonic Ectoderm (ExE) and plays a crucial role in trophoblast development. Previous studies showed that Esrrb is also required to maintain trophoblast stem (TS) cells, the in vitro stem cell model of the early trophoblast lineage. In order to identify regulatory targets of Esrrb in vivo, we performed microarray analysis of Esrrb-null versus wild-type post-implantation ExE, and identified 30 genes down-regulated in Esrrb-mutants. Among them is Bmp4, which is produced by the ExE and known to be critical for primordial germ cell (PGC) specification in vivo. We further identified an enhancer region bound by Esrrb at the Bmp4 locus by performing Esrrb ChIP-seq and luciferase reporter assay using TS cells. Finally, we established a knockout mouse line in which the enhancer region was deleted using CRISPR/Cas9 technology. Both Esrrb-null embryos and enhancer knockout embryos expressed lower levels of Bmp4 in the ExE, and had reduced numbers of PGCs. These results suggested that Esrrb functions as an upstream factor of Bmp4 in the ExE, regulating proper PGC development in mice.
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BMP signaling induces visceral endoderm differentiation of XEN cells and parietal endoderm.
Developmental biology, 2011Co-Authors: Agnieszka Paca, Janet Rossant, Cheryle A Seguin, Melanie Clements, Michael Ryczko, Tristan A. Rodriguez, Tilo KunathAbstract:The Extraembryonic endoderm of mammals is essential for nutritive support of the fetus and patterning of the early embryo. Visceral and parietal endoderm are major subtypes of this lineage with the former exhibiting most, if not all, of the embryonic patterning properties. Extraembryonic endoderm (XEN) cell lines derived from the primitive endoderm of mouse blastocysts represent a cell culture model of this lineage, but are biased towards parietal endoderm in culture and in chimeras. In an effort to promote XEN cells to adopt visceral endoderm character we have mimicked different aspects of the in vivo environment. We found that BMP signaling promoted a mesenchymal-to-epithelial transition of XEN cells with up-regulation of E-cadherin and down-regulation of vimentin. Gene expression analysis showed the differentiated XEN cells most resembled Extraembryonic visceral endoderm (exVE), a subtype of VE covering the Extraembryonic Ectoderm in the early embryo, and during gastrulation it combines with Extraembryonic mesoderm to form the definitive yolk sac. We found that laminin, a major component of the extracellular matrix in the early embryo, synergised with BMP to promote highly efficient conversion of XEN cells to exVE. Inhibition of BMP signaling with the chemical inhibitor, Dorsomorphin, prevented this conversion suggesting that Smad1/5/8 activity is critical for exVE induction of XEN cells. Finally, we show that applying our new culture conditions to freshly isolated parietal endoderm (PE) from Reichert's membrane promoted VE differentiation showing that the PE is developmentally plastic and can be reprogrammed to a VE state in response to BMP. Generation of visceral endoderm from XEN cells uncovers the true potential of these blastocyst-derived cells and is a significant step towards modelling early developmental events ex vivo.
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diethylstilbestrol regulates trophoblast stem cell differentiation as a ligand of orphan nuclear receptor err beta
Genes & Development, 2001Co-Authors: Gilles Tremblay, Janet Rossant, Tilo Kunath, Denis Bergeron, Line Lapointe, Celine Champigny, Joann Bader, Vincent GiguereAbstract:The orphan nuclear receptor ERR beta is expressed in undifferentiated trophoblast stem cell lines and Extraembryonic Ectoderm, and genetic ablation of ERR beta results in abnormal trophoblast proliferation and precocious differentiation toward the giant cell lineage. Here, we show that the synthetic estrogen diethylstilbestrol (DES) promotes coactivator release from ERR beta and inhibits its transcriptional activity. Strikingly, treatment of trophoblast stem cells with DES led to their differentiation toward the polyploid giant cell lineage. In addition, DES-treated pregnant mice exhibited abnormal early placenta development associated with an overabundance of trophoblast giant cells and an absence of diploid trophoblast. These results define a novel pathway for DES action and provide evidence for steroidlike control of trophoblast development.
