Project description:The lateral plate mesoderm (LPM) is a transient tissue that produces a diverse range of differentiated structures, including the limbs. However, the molecular mechanisms that drive early LPM specification and development are poorly understood. In this study, we utilize single-cell transcriptomics to define the cell-fate decisions directing LPM specification, subdivision, and early initiation of the forelimb mesenchyme in chicken embryos. We establish a transcriptional atlas and global cell-cell signalling interactions in progenitor, transitional and mature cell types throughout the developing forelimb field.
Project description:The lung mesenchyme plays important roles in lung development and is affected in many respiratory diseases, yet relatively little is known about the biology of lung mesenchymal progenitors. We sought to establish an induced pluripotent stem cell (iPSC)-based model to study lung mesenchyme development and epithelial-mesenchymal interactions. We generated a mouse iPSC line carrying a lung mesenchyme-specific reporter/tracer to establish a protocol for differentiation into lung mesenchymal progenitors. We derived lung mesenchyme from iPSCs both by directed differentiation via a lateral plate mesodermal progenitor state (induced lung mesenchyme, iLM), and by co-development during lung epithelial differentiation (co-developed lung mesenchyme, cLM). We found that directed differentiation via a lateral plate mesoderm progenitor was not only more efficient, but also yielded engineered lung mesenchymal cells that were more similar to primary lung mesenchyme from day 12.5 mouse embryos, as determined by single cell RNAseq. Our iPSC-derived population will provide an inexhaustible source of cells for studying lung development, modeling diseases, and developing therapeutics.
Project description:The vertebrate forelimb initiates as a localized swelling in the somatic lateral plate mesoderm (somatic LPM) in response to Tbx5-dependent transcription. The molecular pathways driving limb morphogenesis have been extensively studied but the steps directly preceding limb bud formation remain poorly characterized. To address this, we defined the temporal onset of forelimb initiation in mouse embryos using sequencing based high-throughput approaches (RNA-seq, scRNA-seq, ChIP-seq, and Ribo-ITP) benchmarked to known features in forelimb development, identifying four distinct stages. Using scRNA-seq at the onset of forelimb-specific transcription, we determined the transcriptional profile of the somatic LPM and identified signature genes that distinguish the nascent forelimb from other cell types. This group includes multiple genes involved in neural projection as well as cell adhesion. Interestingly, these genes are highly enriched for TBX5 binding sites, suggesting they are candidate early transcriptional targets of TBX5. As TBX5 is essential for forelimb outgrowth, the identification of these genes suggests new mechanistic models for TBX5-driven limb initiation.
Project description:The vertebrate forelimb initiates as a localized swelling in the somatic lateral plate mesoderm (somatic LPM) in response to Tbx5-dependent transcription. The molecular pathways driving limb morphogenesis have been extensively studied but the steps directly preceding limb bud formation remain poorly characterized. To address this, we defined the temporal onset of forelimb initiation in mouse embryos using sequencing based high-throughput approaches (RNA-seq, scRNA-seq, ChIP-seq, and Ribo-ITP) benchmarked to known features in forelimb development, identifying four distinct stages. Using scRNA-seq at the onset of forelimb-specific transcription, we determined the transcriptional profile of the somatic LPM and identified signature genes that distinguish the nascent forelimb from other cell types. This group includes multiple genes involved in neural projection as well as cell adhesion. Interestingly, these genes are highly enriched for TBX5 binding sites, suggesting they are candidate early transcriptional targets of TBX5. As TBX5 is essential for forelimb outgrowth, the identification of these genes suggests new mechanistic models for TBX5-driven limb initiation.
Project description:Renal precusors of the Xenopus pronephros arise from dorso-lateral mesoderm at the early neurula stage. This process is under the control of retinoic acid (RA). We have used microarrays to identify RA targets in dorso-latearl mesoderm by performing differential expression analysis between control and RA-depleted situations
Project description:The data revealed differential expression between floor plate and ventral lateral region in E10.5 mouse embryo midbrain. Several differentially expressed genes in these regions have been reported in the literature, demonstrating reliability of tissue dissection. Midbrain floor plate and non-overlapping adjacent ventral lateral region of mouse E10.5 embryo midbrain was dissected. Each sample was a pool from 6 embryos. Three replicates for each region were used for the experiment.