PROJECT ID: GSE144944

Data source GEO: GSE144944
Description In order to study FGFR3’s role in cranial vault development, we generated the first fgfr3 loss-of-function zebrafish (fgfr3lof1/lof1). The mutant fish exhibited major changes in the craniofacial skeleton, with a lack of sutures, abnormal frontal and parietal bones, and the presence of ectopic bones. In order to define cellular and molecular processes responsible for this phenotype, we performed single-cell RNA sequencing of cranial vault cells from fgfr3lof1/lof1 and fgfr3+/+ zebrafish. Our data allow us to define transcriptomic profile of each osteogenic subpopulation involved in cranial vault formation and we revealed a defect in osteoblast differentiation associated with changes in the extracellular matrix. These findings demonstrate that fgfr3 is a positive regulator of osteogenesis and we conclude that changes in the extracellular matrix within growing bone impair cell-cell communication, mineralization, and new osteoblast recruitment.
Key word fgfr3; cranial vault defects; osteogenesis; osteoblasts; growth-factor receptor-3; osteogenic differentiation; extracellular-matrix; bone-formation; wormian bones; tall stature; mutation; expression; craniosynostosis; protein
Publication Dambroise E, Ktorza I, Brombin A, Abdessalem G et al. Fgfr3 Is a Positive Regulator of Osteoblast Expansion and Differentiation During Zebrafish Skull Vault Development. J Bone Miner Res 2020 Sep;35(9):1782-1797. PMID: 32379366
Abstract Gain or loss-of-function mutations in fibroblast growth factor receptor 3 (FGFR3) result in cranial vault defects highlighting the protein's role in membranous ossification. Zebrafish express high levels of fgfr3 during skull development; in order to study FGFR3's role in cranial vault development, we generated the first fgfr3 loss-of-function zebrafish (fgfr3( lof/lof)). The mutant fish exhibited major changes in the craniofacial skeleton, with a lack of sutures, abnormal frontal and parietal bones, and the presence of ectopic bones. Integrated analyses (in vivo imaging and single-cell RNA sequencing of the osteoblast lineage) of zebrafish fgfr3( lof/lof) revealed a delay in osteoblast expansion and differentiation, together with changes in the extracellular matrix. These findings demonstrate that fgfr3 is a positive regulator of osteogenesis. We conclude that changes in the extracellular matrix within growing bone might impair cell-cell communication, mineralization, and new osteoblast recruitment. (c) 2020 American Society for Bone and Mineral Research.


Dataset Information

Dataset ID Species Tissue / Organ Experiment type Sample Source dataset ID
1. GSE144944 (fgfr3 +|+) Danio rerio cranial vault baseline 1mpf, fgfr3 +/+ GEO: GSM4301248
2. GSE144944 (fgfr3 -|-) Danio rerio cranial vault growth 1mpf, fgfr3 -/- GEO: GSM4301250

Clustering Result

Cluster Cell type Gene id (symbol) Marker class Evidence
1 Osteoblast ENSDARG00000043806 (postna) marker DOI:10.1002/jbmr.4042
1 Osteoblast ENSDARG00000093303 (ifitm1) marker DOI:10.1002/jbmr.4042
2 Chondrocytes ENSDARG00000011407 (col2a1b) marker DOI:10.1002/jbmr.4042
3 Chondrocytes ENSDARG00000011407 (col2a1b) marker DOI:10.1002/jbmr.4042
4 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1016/j.celrep.2019.04.030
4 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1038/nature25969
5 Chondrocytes ENSDARG00000011407 (col2a1b) marker DOI:10.1002/jbmr.4042
6 Osteoblast ENSDARG00000019516 (sp7) marker DOI:10.1002/jbmr.4042
6 Osteoblast ENSDARG00000044276 (spp1) marker DOI:10.1002/jbmr.4042
6 Osteoblast ENSDARG00000058414 (bglap) marker DOI:10.1002/jbmr.4042
6 Osteoblast ENSDARG00000101535 (col10a1b) marker DOI:10.1002/jbmr.4042
7 Epidermal cells ENSDARG00000102750 (cdh1) marker DOI:10.1002/jbmr.4042
8 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1016/j.celrep.2019.04.030
8 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1038/nature25969
9 Epidermal cells ENSDARG00000040534 (epcam) marker DOI:10.1002/jbmr.4042
10 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1002/jbmr.4042
10 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1002/jbmr.4042
11 Epidermal cells ENSDARG00000102750 (cdh1) marker DOI:10.1002/jbmr.4042
12 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1016/j.celrep.2019.04.030
12 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1038/nature25969
13 Epidermal cells ENSDARG00000102750 (cdh1) marker DOI:10.1002/jbmr.4042
14 Neural crest cells ENSDARG00000021032 (foxd3) marker DOI:10.1002/jbmr.4042
15 Oligodendrocytes ENSDARG00000036186 (mbpa) marker DOI:10.1002/jbmr.4042
16 Periderm cells ENSDARG00000057437 (apodb) marker DOI:10.1002/jbmr.4042
17 Pigment cells ENSDARG00000003732 (mitfa) marker DOI:10.1002/jbmr.4042
17 Pigment cells ENSDARG00000057575 (pnp4a) marker DOI:10.1002/jbmr.4042
17 Pigment cells ENSDARG00000062688 (gpnmb) marker DOI:10.1002/jbmr.4042
17 Pigment cells ENSDARG00000025912 (bscl2l) marker DOI:10.1002/jbmr.4042
18 Macrophages ENSDARG00000077809 (il13) marker DOI:10.1002/jbmr.4042
19 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1038/nature25969
19 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1016/j.celrep.2019.04.030
20 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1016/j.celrep.2019.04.030
20 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1038/nature25969
21 Endothelium ENSDARG00000054632 (fli1a) marker DOI:10.1016/j.ydbio.2006.08.020
21 Endothelium ENSDARG00000105215 (kdrl) marker DOI:10.1016/j.ydbio.2006.08.020
21 Endothelium ENSDARG00000004105 (tie1) marker DOI:10.1016/j.ydbio.2006.08.020
21 Endothelium ENSDARG00000077039 (esama) marker DOI:10.1016/j.ydbio.2006.08.020

Cluster Cell type Gene id (symbol) Marker class Evidence
1 Osteoblast ENSDARG00000043806 (postna) marker DOI:10.1002/jbmr.4042
1 Osteoblast ENSDARG00000093303 (ifitm1) marker DOI:10.1002/jbmr.4042
2 Chondrocytes ENSDARG00000011407 (col2a1b) marker DOI:10.1002/jbmr.4042
3 Osteoblast ENSDARG00000043806 (postna) marker DOI:10.1002/jbmr.4042
3 Osteoblast ENSDARG00000093303 (ifitm1) marker DOI:10.1002/jbmr.4042
4 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1016/j.celrep.2019.04.030
4 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1038/nature25969
5 Epidermal cells ENSDARG00000102750 (cdh1) marker DOI:10.1002/jbmr.4042
5 Epidermal cells ENSDARG00000040534 (epcam) marker DOI:10.1002/jbmr.4042
6 Chondrocytes ENSDARG00000011407 (col2a1b) marker DOI:10.1002/jbmr.4042
7 Osteoblast ENSDARG00000019516 (sp7) marker DOI:10.1002/jbmr.4042
7 Osteoblast ENSDARG00000044276 (spp1) marker DOI:10.1002/jbmr.4042
7 Osteoblast ENSDARG00000058414 (bglap) marker DOI:10.1002/jbmr.4042
7 Osteoblast ENSDARG00000101535 (col10a1b) marker DOI:10.1002/jbmr.4042
8 Epidermal cells ENSDARG00000040534 (epcam) marker DOI:10.1002/jbmr.4042
8 Epidermal cells ENSDARG00000102750 (cdh1) marker DOI:10.1002/jbmr.4042
9 Central support cells ENSDARG00000009094 (gata2b) marker DOI:10.1002/jbmr.4042
11 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1016/j.celrep.2019.04.030
11 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1038/nature25969
12 Epidermal cells ENSDARG00000102750 (cdh1) marker DOI:10.1002/jbmr.4042
12 Epidermal cells ENSDARG00000040534 (epcam) marker DOI:10.1002/jbmr.4042
14 Oligodendrocytes ENSDARG00000036186 (mbpa) marker DOI:10.1002/jbmr.4042
15 Epidermal cells ENSDARG00000102750 (cdh1) marker DOI:10.1002/jbmr.4042
15 Epidermal cells ENSDARG00000040534 (epcam) marker DOI:10.1002/jbmr.4042
16 Periderm cells ENSDARG00000057437 (apodb) marker DOI:10.1002/jbmr.4042
17 Neuron ENSDARG00000077799 (egr4) marker DOI:10.1002/jbmr.4042
19 Pigment cells ENSDARG00000003732 (mitfa) marker DOI:10.1002/jbmr.4042
19 Pigment cells ENSDARG00000057575 (pnp4a) marker DOI:10.1002/jbmr.4042
19 Pigment cells ENSDARG00000062688 (gpnmb) marker DOI:10.1002/jbmr.4042
20 Immune cells ENSDARG00000055186 (ccr9a) marker DOI:10.1016/j.celrep.2019.04.030
20 Immune cells ENSDARG00000041959 (cxcr4b) marker DOI:10.1038/nature25969