Craniofacial morphogenesis depends on complex cell fate decisions during the differentiation of post-migratory cranial neural crest cells. Molecular mechanisms of cell differentiation of mesenchymal cells to developing bones, cartilage, teeth, tongue, and other craniofacial tissues are still poorly understood. We performed single-cell transcriptomic analysis of craniofacial mesenchymal cells derived from cranial NCCs in mouse embryo. Using FACS sorting of Wnt1-Cre2 progeny, we carefully mapped the cell heterogeneity in the craniofacial region during the initial stages of cartilage and bone formation. Transcriptomic data and in vivo validations identified molecular determinants of major cell populations involved in the development of lower and upper jaw, teeth, tongue, dermis, or periocular mesenchyme. Single-cell transcriptomic analysis of Meis2-deficient mice revealed critical gene expression differences, including increased osteogenic and cell adhesion markers. This leads to affected mesenchymal cell differentiation and increased ossification, resulting in impaired bone, cartilage, and tongue formation.

Department of Developmental Biology Institute of Experimental Medicine Czech Academy of Sciences Videnska 1083 14200 Prague Czech Republic

Department of Developmental Biology Institute of Experimental Medicine Czech Academy of Sciences Videnska 1083 14200 Prague Czech Republic; Department of Cell Biology Faculty of Science Charles University Vinicna 7 12000 Prague Czech Republic

Laboratory of Gene Expression Institute of Biotechnology Czech Academy of Sciences Prumyslova 595 25200 Vestec Czech Republic

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