Expression and function of Ebf1 gene during chondrogenesis in chick embryo limb buds

El-Magd, Mohammed A.; Abdelfattah‐Hassan, Ahmed; Elsisy, Rasha A.; Hawsawi, Yousef M.; Oyouni, Atif Abdulwahab A.; Al‐Amer, Osama; El‐Shetry, Eman S.

doi:10.1016/j.gene.2021.145895

Cited by 5 publications

(6 citation statements)

References 35 publications

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“…Simultaneously, SOX9, COL2A1 and AP are significantly downregulated (82). Consistent with these observations, EBF1 gene silencing significantly downregulated SOX9 and ACAN expression in our model (although at different time points).…”

Section: Transient Gene Silencing Of Atoh8 or Ebf1 Blocks Chondrogenesissupporting

confidence: 90%

The temporal transcriptomic signature of cartilage formation

Takács

Vágó

Póliska

et al. 2022

Preprint

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Chondrogenesis is a multistep process whereby cartilage progenitor cells generate a tissue with distinct structural and functional properties. Although several approaches to cartilage regeneration rely on the differentiation of implanted progenitor cells, the temporal transcriptomic landscape of in vitro chondrogenesis in different models has not been reported. Using RNA sequencing, we examined the differences between gene expression patterns during early cartilage formation in micromass cultures of embryonic limb bud-derived progenitors. Principal component analysis indicated a progressively different and distinct transcriptome during chondrogenesis. Differentially expressed genes (DEGs) based on pairwise comparisons of samples from consecutive days were classified into clusters and analysed. We confirmed the involvement of the top DEGs in chondrogenic differentiation with pathway analysis. We discovered several chondrogenesis-associated transcription factors and new collagen subtypes not previously linked to cartilage formation. These results provide, for the first time, a detailed insight into the molecular mechanism of in vitro chondrogenesis and provide a new gold standard for the temporal transcriptomic landscape of chondrogenic differentiation that may serve as a platform for new approaches in cartilage tissue engineering.

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Section: Transient Gene Silencing Of Atoh8 or Ebf1 Blocks Chondrogenesissupporting

confidence: 90%

The temporal transcriptomic signature of cartilage formation

Takács

Vágó

Póliska

et al. 2022

Preprint

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“…Cell types assigned to Clusters 10 and 12 were annotated as myocyte progenitors or myocytes based on their expression of Ttn , Myog , Myod1 , Myf5 , and Tnnt1 ( Figure 3 ; Supplementary Table S2 ) ( Tajbakhsh et al, 1996 ; Beauchamp et al, 2000 ; Linke and Kruger, 2010 ; Wood et al, 2013 ; Ganassi et al, 2018 ; Li et al, 2021 ; Yagi et al, 2021 ). Cell types assigned to Cluster 6 were annotated as joint interzone cells based on their expression of Ebf1 , Jun , Hoxd13 , and Gdf5 ( Figure 3 , Supplementary Table S2 ) ( Storm and Kingsley, 1999 ; Kan and Tabin, 2013 ; Huang et al, 2016 ; Shwartz et al, 2016 ; Capellini et al, 2017 ; El-Magd et al, 2021 ). Cell types assigned to Cluster 11 were annotated as interdigital mesenchyme based on their significant expression of several interdigital tissue markers including: Hoxa13 , Hoxd13 , Msx1 , Wnt5a , and Aldh1a2 ( Figure 3 ; Supplementary Figure S3 ).…”

Section: Resultsmentioning

confidence: 99%

Digits in a dish: An in vitro system to assess the molecular genetics of hand/foot development at single-cell resolution

Fuiten

Yoshimoto

Shukunami

et al. 2023

Front. Cell Dev. Biol.

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In vitro models allow for the study of developmental processes outside of the embryo. To gain access to the cells mediating digit and joint development, we identified a unique property of undifferentiated mesenchyme isolated from the distal early autopod to autonomously re-assemble forming multiple autopod structures including: digits, interdigital tissues, joints, muscles and tendons. Single-cell transcriptomic analysis of these developing structures revealed distinct cell clusters that express canonical markers of distal limb development including: Col2a1, Col10a1, and Sp7 (phalanx formation), Thbs2 and Col1a1 (perichondrium), Gdf5, Wnt5a, and Jun (joint interzone), Aldh1a2 and Msx1 (interdigital tissues), Myod1 (muscle progenitors), Prg4 (articular perichondrium/articular cartilage), and Scx and Tnmd (tenocytes/tendons). Analysis of the gene expression patterns for these signature genes indicates that developmental timing and tissue-specific localization were also recapitulated in a manner similar to the initiation and maturation of the developing murine autopod. Finally, the in vitro digit system also recapitulates congenital malformations associated with genetic mutations as in vitro cultures of Hoxa13 mutant mesenchyme produced defects present in Hoxa13 mutant autopods including digit fusions, reduced phalangeal segment numbers, and poor mesenchymal condensation. These findings demonstrate the robustness of the in vitro digit system to recapitulate digit and joint development. As an in vitro model of murine digit and joint development, this innovative system will provide access to the developing limb tissues facilitating studies to discern how digit and articular joint formation is initiated and how undifferentiated mesenchyme is patterned to establish individual digit morphologies. The in vitro digit system also provides a platform to rapidly evaluate treatments aimed at stimulating the repair or regeneration of mammalian digits impacted by congenital malformation, injury, or disease.

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“…EOM progenitors were characterised by a unique signature of regulons involved in connective tissue/ECM remodelling including Egr1 (Havis and Duprez, 2020b) and Creb3l1, a downstream effector of Thr signaling (García et al, 2017) that plays a critical role during bone development and activates extracellular matrix genes such as Col1a1 and Fn1 (Sampieri et al, 2019). Top regulons of EOM progenitors are also involved in cell proliferation ( Foxc1 (Yang et al, 2017), Sox4 (Moreno, 2019), Fos (Almada et al, 2021), Klf6 (Dionyssiou et al, 2013), Ebf1 (Györy et al, 2012)), commitment into endothelial and smooth muscle fates ( Foxc1 (Han et al, 2017; Mayeuf-Louchart et al, 2016; Whitesell et al, 2019a; Yang et al, 2017)) as well as differentiation into mesenchymal lineages such as pericytes, adipocytes and chondrocytes (Ebf1, (El-Magd et al, 2021; Jimenez et al, 2007; Pagani et al, 2021). Notably, some of these regulons were also active in the EOM during development and implicated in non-myogenic cell fates from bipotent myogenic cells (Grimaldi et al, 2022).…”

Section: Resultsmentioning

confidence: 99%

Extraocular muscle stem cells exhibit distinct cellular properties associated with non-muscle molecular signatures

Girolamo

Benavente-Diaz

Grimaldi

et al. 2023

Preprint

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The muscle stem cell (MuSC) population is recognized as functionally heterogeneous. Cranial muscle stem cells, which originate from head mesoderm, can have greater proliferative capacity in culture and higher regenerative potential in transplantation assays when compared to those in the limb. The existence of such functional differences in phenotypic outputs remain unresolved as a comprehensive understanding of the underlying mechanisms is lacking. We addressed this issue using a combination of clonal analysis, live imaging, and scRNA-seq, identifying critical biological features that distinguish extraocular (EOM) and limb (Tibialis anterior, TA) MuSC populations. Time-lapse studies using a MyogenintdTomato reporter showed that the increased proliferation capacity of EOM MuSCs is accompanied by a differentiation delay in vitro. Unexpectedly, in vitro activated EOM MuSCs expressed a large array of distinct extracellular matrix (ECM) components, growth factors, and signaling molecules that are typically associated with mesenchymal non-muscle cells. These unique features are regulated by a specific set of transcription factors that constitute a coregulating module. This transcription factor network, which includes Foxc1 as one of the major players, appears to be hardwired to EOM identity as it is present in quiescent adult MuSCs, in the activated counterparts during growth and retained upon passages in vitro. These findings provide insights into how high-performing MuSCs regulate myogenic commitment by active remodeling of their local environment.

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Expression and function of Ebf1 gene during chondrogenesis in chick embryo limb buds

Cited by 5 publications

References 35 publications

The temporal transcriptomic signature of cartilage formation

The temporal transcriptomic signature of cartilage formation

Digits in a dish: An in vitro system to assess the molecular genetics of hand/foot development at single-cell resolution

Extraocular muscle stem cells exhibit distinct cellular properties associated with non-muscle molecular signatures

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