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Scientific Reports 2012Directed specification and prospective isolation of chondrogenic paraxial mesoderm progeny from human pluripotent stem (PS) cells have not yet been achieved. Here we...
Directed specification and prospective isolation of chondrogenic paraxial mesoderm progeny from human pluripotent stem (PS) cells have not yet been achieved. Here we report the successful generation of KDR(-)PDGFRα(+) progeny expressing paraxial mesoderm genes and the mesendoderm reporter MIXL1-GFP in a chemically defined medium containing the canonical WNT signaling activator, BMP-inhibitor, and the Nodal/Activin/TGFβ signaling controller. Isolated (GFP(+))KDR(-)PDGFRα(+) mesoderm cells were sensitive to sequential addition of the three chondrogenic factors PDGF, TGFβ and BMP. Under these conditions, the cells showed robust chondrogenic activity in micromass culture, and generated a hyaline-like translucent cartilage particle in serum-free medium. In contrast, both STRO1(+) mesenchymal stem/stromal cells from adult human marrow and mesenchymal cells spontaneously arising from hPS cells showed a relatively weaker chondrogenic response in vitro, and formed more of the fibrotic cartilage particles. Thus, hPS cell-derived KDR(-)PDGFRα(+ )paraxial mesoderm-like cells have potential in engineered cartilage formation and cartilage repair.
Topics: Adult; Animals; Antigens, Surface; Basic Helix-Loop-Helix Transcription Factors; Bone Morphogenetic Proteins; Cell Differentiation; Cells, Cultured; Chondrocytes; Chondrogenesis; Embryonic Stem Cells; Flow Cytometry; Gene Expression Profiling; Green Fluorescent Proteins; Homeodomain Proteins; Humans; Mesoderm; Mice; Microscopy, Fluorescence; Platelet-Derived Growth Factor; Pluripotent Stem Cells; Prospective Studies; Receptor, Platelet-Derived Growth Factor alpha; Transcription Factors; Transforming Growth Factor beta; Vascular Endothelial Growth Factor Receptor-2
PubMed: 22701159
DOI: 10.1038/srep00455 -
Annals of the New York Academy of... Jun 1996
Review
Topics: Amino Acid Sequence; Animals; Bone and Bones; Cartilage; Chick Embryo; Consensus Sequence; Embryonic and Fetal Development; Gene Expression Regulation, Developmental; Helix-Loop-Helix Motifs; Mesoderm; Molecular Sequence Data; Sequence Homology, Amino Acid; Transcription Factors; Vertebrates
PubMed: 8702115
DOI: 10.1111/j.1749-6632.1996.tb56248.x -
Nature Reviews. Molecular Cell Biology Nov 2014Segmentation of the paraxial mesoderm is a major event of vertebrate development that establishes the metameric patterning of the body axis. This process involves the... (Review)
Review
Segmentation of the paraxial mesoderm is a major event of vertebrate development that establishes the metameric patterning of the body axis. This process involves the periodic formation of sequential units, termed somites, from the presomitic mesoderm. Somite formation relies on a molecular oscillator, the segmentation clock, which controls the rhythmic activation of several signalling pathways and leads to the oscillatory expression of a subset of genes in the presomitic mesoderm. The response to the periodic signal of the clock, leading to the establishment of the segmental pre-pattern, is gated by a system of travelling signalling gradients, often referred to as the wavefront. Recent studies have advanced our understanding of the molecular mechanisms involved in the generation of oscillations and how they interact and are coordinated to activate the segmental gene expression programme.
Topics: Animals; Biological Clocks; Body Patterning; CLOCK Proteins; Embryonic Development; Gene Expression Regulation, Developmental; Humans; Mesoderm; Models, Biological; Receptors, Notch; Signal Transduction; Vertebrates; Wnt Proteins
PubMed: 25335437
DOI: 10.1038/nrm3891 -
Cell Cycle (Georgetown, Tex.) Jul 2020It is hard to supply satellite cells as a cell source for therapy of muscle degenerative disease since the sampling of muscle tissue is very invasive to a patient with...
It is hard to supply satellite cells as a cell source for therapy of muscle degenerative disease since the sampling of muscle tissue is very invasive to a patient with muscular disease. Direct conversion allows us to get specific cell types by transduction of defined transcriptional factors. To induce myogenic direct conversion, we transduced five transcriptional factors including Pax3, Sox2, Klf4, c-Myc, and Esrrb into mouse embryonic fibroblasts. We found that the transduction of the five transcriptional factors induced myogenic direct conversion of fibroblast. We revealed that the transduced cells with the five transcriptional factors were converted to myogenic lineage cells through a paraxial mesoderm-like stage. The expression level of myogenic-related genes of the transduced cells gradually increased as the passage increased. The induced myogenic lineage cells differentiated into muscle fibers and . The current study revealed that the five transcription factors generated myogenic lineage cells from fibroblast via a paraxial mesoderm stage. The induced myogenic lineage cells may have a potential being applied as cell source for degenerative muscle disease.
Topics: Animals; Biomarkers; Cell Differentiation; Cell Lineage; Cellular Reprogramming; Embryo, Mammalian; Fibroblasts; Gene Expression Regulation, Developmental; Kruppel-Like Factor 4; Mesoderm; Mice; Muscle Cells; Muscle Development; Muscle Fibers, Skeletal; Stem Cells; Transcription Factors
PubMed: 32579865
DOI: 10.1080/15384101.2020.1780384 -
The International Journal of... 2017The vertebrate head characteristically exhibits a complex pattern with sense organs, brain, paired eyes and jaw muscles, and the brain case is not found in other... (Review)
Review
The vertebrate head characteristically exhibits a complex pattern with sense organs, brain, paired eyes and jaw muscles, and the brain case is not found in other chordates. How the extant vertebrate head has evolved remains enigmatic. Historically, there have been two conflicting views on the origin of the vertebrate head, segmental and non-segmental views. According to the segmentalists, the vertebrate head is organized as a metameric structure composed of segments equivalent to those in the trunk; a metamere in the vertebrate head was assumed to consist of a somite, a branchial arch and a set of cranial nerves, considering that the head evolved from rostral segments of amphioxus-like ancestral vertebrates. Non-segmentalists, however, considered that the vertebrate head was not segmental. In that case, the ancestral state of the vertebrate head may be non-segmented, and rostral segments in amphioxus might have been secondarily gained, or extant vertebrates might have evolved through radical modifications of amphioxus-like ancestral vertebrate head. Comparative studies of mesodermal development in amphioxus and vertebrate gastrula embryos have revealed that mesodermal gene expressions become segregated into two domains anteroposteriorly to specify the head mesoderm and trunk mesoderm only in vertebrates; in this segregation, key genes such as delta and hairy, involved in segment formation, are expressed in the trunk mesoderm, but not in the head mesoderm, strongly suggesting that the head mesoderm of extant vertebrates is not segmented. Taken together, the above finding possibly adds a new insight into the origin of the vertebrate head; the vertebrate head mesoderm would have evolved through an anteroposterior polarization of the paraxial mesoderm if the ancestral vertebrate had been amphioxus-like.
Topics: Animals; Body Patterning; Cephalochordata; Gene Expression Regulation, Developmental; Head; Lancelets; Models, Biological; Somites; Vertebrates
PubMed: 29319111
DOI: 10.1387/ijdb.170121to -
Development (Cambridge, England) Mar 2018Body skeletal muscles derive from the paraxial mesoderm, which forms in the posterior region of the embryo. Using microarrays, we characterize novel mouse presomitic...
Body skeletal muscles derive from the paraxial mesoderm, which forms in the posterior region of the embryo. Using microarrays, we characterize novel mouse presomitic mesoderm (PSM) markers and show that, unlike the abrupt transcriptome reorganization of the PSM, neural tube differentiation is accompanied by progressive transcriptome changes. The early paraxial mesoderm differentiation stages can be efficiently recapitulated using mouse and human pluripotent stem cells. While Wnt activation alone can induce posterior PSM markers, acquisition of a committed PSM fate and efficient differentiation into anterior PSM Pax3 identity further requires BMP inhibition to prevent progenitors from drifting to a lateral plate mesoderm fate. When transplanted into injured adult muscle, these precursors generated large numbers of immature muscle fibers. Furthermore, exposing these mouse PSM-like cells to a brief FGF inhibition step followed by culture in horse serum-containing medium allows efficient recapitulation of the myogenic program to generate myotubes and associated Pax7 cells. This protocol results in improved differentiation and maturation of mouse muscle fibers over serum-free protocols and enables the study of myogenic cell fusion and satellite cell differentiation.
Topics: Animals; Bone Morphogenetic Proteins; Cell Differentiation; Flow Cytometry; Gene Expression Profiling; Gene Expression Regulation, Developmental; Humans; Immunohistochemistry; Immunophenotyping; In Situ Hybridization; In Vitro Techniques; Mesoderm; Mice; Muscle Development; Muscle, Skeletal; Pluripotent Stem Cells; Real-Time Polymerase Chain Reaction; Tissue Array Analysis; Wnt Signaling Pathway
PubMed: 29555813
DOI: 10.1242/dev.157339 -
Developmental Biology Jul 2004During vertebrate embryogenesis, the newly formed mesoderm is allocated to the paraxial, intermediate, and lateral domains, each giving rise to different cell and tissue... (Comparative Study)
Comparative Study
During vertebrate embryogenesis, the newly formed mesoderm is allocated to the paraxial, intermediate, and lateral domains, each giving rise to different cell and tissue types. Here, we provide evidence that the forkhead genes, Foxc1 and Foxc2, play a role in the specification of mesoderm to paraxial versus intermediate fates. Mouse embryos lacking both Foxc1 and Foxc2 show expansion of intermediate mesoderm markers into the paraxial domain, lateralization of somite patterning, and ectopic and disorganized mesonephric tubules. In gain of function studies in the chick embryo, Foxc1 and Foxc2 negatively regulate intermediate mesoderm formation. By contrast, their misexpression in the prospective intermediate mesoderm appears to drive cells to acquire paraxial fate, as revealed by expression of the somite markers Pax7 and Paraxis. Taken together, the data indicate that Foxc1 and Foxc2 regulate the establishment of paraxial versus intermediate mesoderm cell fates in the vertebrate embryo.
Topics: Animals; Basic Helix-Loop-Helix Transcription Factors; Body Patterning; Cell Differentiation; Chick Embryo; DNA-Binding Proteins; Electroporation; Embryo, Mammalian; Forkhead Transcription Factors; Gene Expression Regulation, Developmental; Homeodomain Proteins; Immunohistochemistry; In Situ Hybridization; Mesoderm; Mice; PAX7 Transcription Factor; Somites; Transcription Factors
PubMed: 15196959
DOI: 10.1016/j.ydbio.2004.03.034 -
Nature Communications Feb 2021Somites arising from paraxial mesoderm are a hallmark of the segmented vertebrate body plan. They form sequentially during axis extension and generate musculoskeletal...
Somites arising from paraxial mesoderm are a hallmark of the segmented vertebrate body plan. They form sequentially during axis extension and generate musculoskeletal cell lineages. How paraxial mesoderm becomes regionalised along the axis and how this correlates with dynamic changes of chromatin accessibility and the transcriptome remains unknown. Here, we report a spatiotemporal series of ATAC-seq and RNA-seq along the chick embryonic axis. Footprint analysis shows differential coverage of binding sites for several key transcription factors, including CDX2, LEF1 and members of HOX clusters. Associating accessible chromatin with nearby expressed genes identifies cis-regulatory elements (CRE) for TCF15 and MEOX1. We determine their spatiotemporal activity and evolutionary conservation in Xenopus and human. Epigenome silencing of endogenous CREs disrupts TCF15 and MEOX1 gene expression and recapitulates phenotypic abnormalities of anterior-posterior axis extension. Our integrated approach allows dissection of paraxial mesoderm regulatory circuits in vivo and has implications for investigating gene regulatory networks.
Topics: Animals; Basic Helix-Loop-Helix Transcription Factors; CDX2 Transcription Factor; Cell Lineage; Chick Embryo; Chromatin; Female; Gastrulation; Gene Expression Regulation, Developmental; Homeodomain Proteins; Lymphoid Enhancer-Binding Factor 1; Mesoderm; Regulatory Sequences, Nucleic Acid; Somites; Transcription Factors; Transcriptome; Xenopus laevis
PubMed: 33608545
DOI: 10.1038/s41467-021-21426-7 -
Developmental Biology Jun 2000A new bHLH gene from mouse that we call pMesogenin1 (referring to paraxial mesoderm-specific expression and regulatory capacities) and its candidate ortholog from...
A new bHLH gene from mouse that we call pMesogenin1 (referring to paraxial mesoderm-specific expression and regulatory capacities) and its candidate ortholog from Xenopus were isolated and studied comparatively. In both organisms the gene is specifically expressed in unsegmented paraxial mesoderm and its immediate progenitors. A striking feature of pMesogenin1 expression is that it terminates abruptly in presumptive somites (somitomeres). Somitomeres rostral to the pMesogenin1 domain strongly upregulate expression of pMesogenin's closest known paralogs, MesP1 and MesP2 (Thylacine1/2 in Xenopus). Subsequently, the most rostral somitomere becomes a new somite and expression of MesP1/2 is sharply downregulated before this transition. Thus, expression patterns of these bHLH genes, together with that of an additional bHLH gene in the mouse, Paraxis, collectively define discrete but highly dynamic prepatterned subdomains of the paraxial mesoderm. In functional assays, we show that pMesogenin1 from either mouse or frog can efficiently drive nonmesodermal cells to assume a phenotype with molecular and cellular characteristics of early paraxial mesoderm. Among genes induced by added pMesogenin1 is Xwnt-8, a signaling factor that induces a similar repertoire of marker genes and a similar cellular phenotype. Additional target genes induced by pMesogenin1 are ESR4/5, regulators known to play a significant role in segmentation of paraxial mesoderm (W. C. Jen et al., 1999, Genes Dev. 13, 1486-1499). pMesogenin1 differs from other known mesoderm-inducing transcription factors because it does not also activate a dorsal (future axial) mesoderm phenotype, suggesting that pMesogenin1 is involved in specifying paraxial mesoderm. In the context of the intact frog embryo, ectopic pMesogenin1 also actively suppressed axial mesoderm markers and disrupted normal formation of notochord. In addition, we found evidence for cross-regulatory interactions between pMesogenin1 and T-box transcription factors, a family of genes normally expressed in a broader pattern and known to induce multiple types of mesoderm. Based on our results and results from prior studies of related bHLH genes, we propose that pMesogenin1 and its closest known relatives, MesP1/2 (in mouse) and Thylacine1/2 (in Xenopus), comprise a bHLH subfamily devoted to formation and segmentation of paraxial mesoderm.
Topics: Amino Acid Sequence; Animals; Basic Helix-Loop-Helix Transcription Factors; Body Patterning; DNA-Binding Proteins; Embryonic and Fetal Development; Gene Expression Regulation, Developmental; Gene Library; Helix-Loop-Helix Motifs; In Situ Hybridization; Mesoderm; Mice; Mice, Inbred Strains; Molecular Sequence Data; Phenotype; Phylogeny; Reverse Transcriptase Polymerase Chain Reaction; Sequence Alignment; Sequence Homology, Amino Acid; Signal Transduction; Transcription Factors; Xenopus laevis
PubMed: 10837126
DOI: 10.1006/dbio.2000.9717 -
Seminars in Cell & Developmental Biology Dec 2017Pluripotent stem cells represent important tools for both basic and translational science as they enable to study mechanisms of development, model diseases in vitro and... (Review)
Review
Pluripotent stem cells represent important tools for both basic and translational science as they enable to study mechanisms of development, model diseases in vitro and provide a potential source of tissue-specific progenitors for cell therapy. Concomitantly with the increasing knowledge of the molecular mechanisms behind activation of the skeletal myogenic program during embryonic development, novel findings in the stem cell field provided the opportunity to begin recapitulating in vitro the events occurring during specification of the myogenic lineage. In this review, we will provide a perspective of the molecular mechanisms responsible for skeletal myogenic commitment in the embryo and how this knowledge was instrumental for specifying this lineage from pluripotent stem cells. In addition, we will discuss the current limitations for properly recapitulating skeletal myogenesis in the petri dish, and we will provide insights about future applications of pluripotent stem cell-derived myogenic cells.
Topics: Animals; Cell Differentiation; Cell Lineage; Gene Expression Regulation, Developmental; Humans; Mesoderm; Muscle Development; Muscle Proteins; Muscle, Skeletal; Pluripotent Stem Cells
PubMed: 29107681
DOI: 10.1016/j.semcdb.2017.10.031