Single-cell analysis identifies a key role for Hhip in murine coronal suture development

• Published in: Nature communications Vol. 12; no. 1; pp. 7132 - 16
• Main Authors: Holmes, Greg, Gonzalez-Reiche, Ana S., Saturne, Madrikha, Motch Perrine, Susan M., Zhou, Xianxiao, Borges, Ana C., Shewale, Bhavana, Richtsmeier, Joan T., Zhang, Bin, van Bakel, Harm, Jabs, Ethylin Wang
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 08.12.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 13/51; 14; 38; 38/32; 38/39; 38/91; 45; 631/136/1425; 631/136/1455; 631/136/818; 631/1647/514/1949; 631/553/2694; Animals; Biomedical materials; Bone Development; Bone growth; Bones; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell Proliferation; Cranial sutures; Cranial Sutures - growth & development; Cranial Sutures - pathology; Craniofacial growth; Craniosynostoses; DNA Topoisomerases, Type II; Embryos; Female; Gene Expression Regulation, Developmental; Hedgehog protein; Hedgehog Proteins - metabolism; Humanities and Social Sciences; Inhibitors; Maintenance; Male; Membrane Glycoproteins - genetics; Membrane Glycoproteins - metabolism; Mesenchyme; Mesoderm; Mice; Mice, Inbred C57BL; multidisciplinary; Neonates; Osteogenesis - genetics; Osteogenesis - physiology; Phenotype; Poly-ADP-Ribose Binding Proteins; Population structure; Populations; Science; Science (multidisciplinary); Sequence Analysis, RNA; Signal Transduction; Signaling; Single-Cell Analysis - methods; Skull; Sutures; Transcriptome
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-27402-5

Craniofacial development depends on formation and maintenance of sutures between bones of the skull. In sutures, growth occurs at osteogenic fronts along the edge of each bone, and suture mesenchyme separates adjacent bones. Here, we perform single-cell RNA-seq analysis of the embryonic, wild type murine coronal suture to define its population structure. Seven populations at E16.5 and nine at E18.5 comprise the suture mesenchyme, osteogenic cells, and associated populations. Expression of Hhip , an inhibitor of hedgehog signaling, marks a mesenchymal population distinct from those of other neurocranial sutures. Tracing of the neonatal Hhip -expressing population shows that descendant cells persist in the coronal suture and contribute to calvarial bone growth. In Hhip −/− coronal sutures at E18.5, the osteogenic fronts are closely apposed and the suture mesenchyme is depleted with increased hedgehog signaling compared to those of the wild type. Collectively, these data demonstrate that Hhip is required for normal coronal suture development. Craniofacial development depends on formation and maintenance of sutures between bones of the skull. Here the authors identify enriched expression of the hedgehog inhibitor Hhip, specifically in the mesenchyme of the murine coronal suture, and show sutural dysgenesis in Hhip−/− mutants.