Sox17 promotes differentiation in mouse embryonic stem cells by directly regulating extraembryonic gene expression and indirectly antagonizing self-renewal

Niakan, Kathy K.; Ji, Hongkai; Maehr, René; Vokes, Steven A.; Rodolfa, Kit T.; Sherwood, Richard I.; Yamaki, Mariko; Dimos, John; Chen, Alice E.; Melton, Douglas A.; McMahon, Andrew P.; Eggan, Kevin

doi:10.1101/gad.1833510

Cited by 272 publications

(309 citation statements)

References 46 publications

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“…Specifically, we note that Sox17 and Foxa2 are also expressed in some embryonic tissues, such as the primitive endoderm, where Brachyury is largely absent (Fig. 3 D and E and Movies S1-S4) (25). Thus, it is likely that these two genes can be regulated by distinct factors and mechanisms at different developmental stages.…”

Section: Brachyury Coordinates Gene Expression During In Vitro Streakmentioning

confidence: 99%

Charting Brachyury-mediated developmental pathways during early mouse embryogenesis

Lolas

Valenzuela

Tjian

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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To gain insights into coordinated lineage-specification and morphogenetic processes during early embryogenesis, here we report a systematic identification of transcriptional programs mediated by a key developmental regulator-Brachyury. High-resolution chromosomal localization mapping of Brachyury by ChIP sequencing and ChIP-exonuclease revealed distinct sequence signatures enriched in Brachyury-bound enhancers. A combination of genomewide in vitro and in vivo perturbation analysis and cross-species evolutionary comparison unveiled a detailed Brachyury-dependent gene-regulatory network that directly links the function of Brachyury to diverse developmental pathways and cellular housekeeping programs. We also show that Brachyury functions primarily as a transcriptional activator genome-wide and that an unexpected gene-regulatory feedback loop consisting of Brachyury, Foxa2, and Sox17 directs proper stem-cell lineage commitment during streak formation. Target gene and mRNA-sequencing correlation analysis of the T c mouse model supports a crucial role of Brachyury in up-regulating multiple key hematopoietic and musclefate regulators. Our results thus chart a comprehensive map of the Brachyury-mediated gene-regulatory network and how it influences in vivo developmental homeostasis and coordination.primitive streak | early development | mesoendoderm differentiation I n the past decade, significant insights have been gained in understanding gene-regulatory programs responsible for embryonic stem (ES) cell pluripotency and self-renewal. However, transcriptional control mechanisms underlying finely balanced lineage-segregation and morphogenetic processes during early mammalian embryogenesis remained elusive. During early mouse gastrulation, Brachyury (T), a classical enhancer-binding transcription factor (TF), has been reported to be required for the proper development of primitive streak, allantois, axial, and posterior mesoderm (reviewed in ref. 1).Specifically, mouse embryos that are homozygous for the Brachyury (T) deletion die at midgestation (2). T −/− mutant epiblast cells are compromised in their ability to migrate away from the primitive streak and, therefore, are unable to undergo the morphogenetic movements carried out by their wild-type (WT) counterparts during gastrulation. In addition to defects in the primitive streak, the notochord is absent in posterior portions of the embryo. Although the anterior portions of T mutant mice contain notochordal precursor-like cells, they fail to undergo normal terminal differentiation (3, 4). The embryonic pattern posterior to the forelimb region of T −/− animals is also disturbed, with somites posterior to the seventh pair absent or abnormal. Although neural folds fuse to form the neural tube, they are severely kinked in the caudal region, and the surface ectoderm tends to form fluid-filled blisters (2, 4, 5). In addition to these well-documented defects, numerous other phenotypic abnormalities have been reported in T −/− and T mutant embryos, including left-right pattern...

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Section: Brachyury Coordinates Gene Expression During In Vitro Streakmentioning

confidence: 99%

Charting Brachyury-mediated developmental pathways during early mouse embryogenesis

Lolas

Valenzuela

Tjian

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…Please click here to view a larger version of this figure. 19,22,27,30,[36][37][38][39][40] have been characterized for the formation of aggregates and, although subtle differences between cell lines are expected and have been observed, all the above lines show similar dynamics in terms of elongation and morphology. In terms of gene expression, the expression pattern is specific to the gene expressed, but within each cell line, the expression pattern is generally consistent over a number of passages.…”

Section: Aggregate Imaging and Quantitative Image Analysismentioning

confidence: 99%

Generation of Aggregates of Mouse Embryonic Stem Cells that Show Symmetry Breaking, Polarization and Emergent Collective Behaviour <em>In Vitro</em>

Baillie-Johnson

Brink

Balayo

et al. 2015

JoVE

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We have developed a protocol improving current Embryoid Body (EB) culture which allows the study of self-organization, symmetry breaking, axial elongation and cell fate specification using aggregates of mouse embryonic stem cells (mESCs) in suspension culture. Small numbers of mESCs are aggregated in basal medium for 48 hr in non-tissue-culture-treated, U-bottomed 96-well plates, after which they are competent to respond to experimental signals. Following treatment, these aggregates begin to show signs of polarized gene expression and gradually alter their morphology from a spherical mass of cells to an elongated, well organized structure in the absence of external asymmetry cues. These structures are not only able to display markers of the three germ layers, but actively display gastrulation-like movements, evidenced by a directional dislodgement of individual cells from the aggregate, which crucially occurs at one region of the elongated structure. This protocol provides a detailed method for the reproducible formation of these aggregates, their stimulation with signals such as Wnt/β-Catenin activation and BMP inhibition and their analysis by single time-point or time-lapse fluorescent microscopy. In addition, we describe modifications to current whole-mount mouse embryo staining procedures for immunocytochemical analysis of specific markers within fixed aggregates. The changes in morphology, gene expression and length of the aggregates can be quantitatively measured, providing information on how signals can alter axial fates. It is envisaged that this system can be applied both to the study of early developmental events such as axial development and organization, and more broadly, the processes of self-organization and cellular decision-making. It may also provide a suitable niche for the generation of cell types present in the embryo that are unobtainable from conventional adherent culture such as spinal cord and motor neurones.

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“…This is an ever-increasing pool that includes GATA binding proteins 4 and 6 [214,215], the Sry-related HMG-box transcription factor Sox7 and Sox17 [216][217][218][219][220], transcription factor 2, Tcf2 (vHnf-1b) [221], FoxA1 (Hnf-3a) [222], FoxA2 (Hnf-3b) [223], Hnf-4 [224] and Disabled-2 [225,226]. These and many more have been identified and validated by microarray analysis of ExEn isolated from embryos [227].…”

Section: Definitive Endoderm Vs the Extraembryonic Endoderm Lineagementioning

confidence: 99%

Retinoic Acid and the Development of the Endoderm

Kelly

Drysdale

2015

JDB

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Retinoic acid (RA) is an important signaling molecule in the development of the endoderm and an important molecule in protocols used to generate endodermal cell types from stem cells. In this review, we describe the RA signaling pathway and its role in the patterning and specification of the extra embryonic endoderm and different endodermal organs. The formation of endoderm is an ancient evolutionary feature and RA signaling appears to have coevolved with the vertebrate lineage. Towards that end, we describe how RA participates in many regulatory networks required for the formation of extraembryonic structures as well as the organs of the embryo proper.

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Sox17 promotes differentiation in mouse embryonic stem cells by directly regulating extraembryonic gene expression and indirectly antagonizing self-renewal

Cited by 272 publications

References 46 publications

Charting Brachyury-mediated developmental pathways during early mouse embryogenesis

Charting Brachyury-mediated developmental pathways during early mouse embryogenesis

Generation of Aggregates of Mouse Embryonic Stem Cells that Show Symmetry Breaking, Polarization and Emergent Collective Behaviour <em>In Vitro</em>

Retinoic Acid and the Development of the Endoderm

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