Abstract
Long bones of the appendicular skeleton are formed through endochondral ossification. Endochondral bone formation initiates with mesenchymal condensation, followed by the formation of a cartilage template which is replaced by bone. Fibroblast growth factor 9 (FGF9) regulates bone development. Fgf9−/− mice exhibit disproportionate shortening of proximal skeletal elements. Fgf9 missense mutations in mice and humans induce joint synostosis. Thus, FGF9 is critical for regulating bone length and joint formation. Conversely, mechanisms regulating bone width remain unclear. Here, we showed that the homozygous elbow knee synostosis (Eks) mutant mice harboring N143T mutation in Fgf9 have wide long bones at birth. We investigated the cellular and molecular mechanisms underlying the widened prospective humerus in Fgf9Eks/Eks embryos. Increased and expanded FGF signaling in concert with wider expression domain of Fgf receptor 3 (Fgfr3) during chondrogenic condensation of the humerus led to widened cartilage, which resulted in the formation of wider prospective humeri in neonatal Fgf9Eks/Eks mice. Increased and expanded FGF signaling during chondrogenic condensation led to increased density of chondrocytes of the humeri accompanied by increased proliferation of chondrocytes which express inappropriately higher levels of cyclin D1 in Fgf9Eks/Eks embryos. The results suggest that FGF9 regulates the width of prospective long bones by controlling the width of chondrogenic condensation.
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Abbreviations
- COL1:
-
Type I collagen
- COL2:
-
Type II collagen
- E:
-
Embryonic day
- EdU:
-
5-Ethynyl- 2′-deoxyuridine
- Eks:
-
Elbow knee synostosis
- FGF:
-
Fibroblast growth factor
- FGFR:
-
Fibroblast growth factor receptor
- HE:
-
Hematoxylin and eosin
- P:
-
Postnatal
- PBS:
-
Phosphate-buffered saline
- pERK1/2:
-
ERK1/2 phosphorylation
- RUNX2:
-
Runt-related transcription factor 2
- SOX9:
-
SRY-box9
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Acknowledgements
We thank H. Koseki for sharing the Eks mouse line. We also thank Enago (www.enago.jp) for English language editing. This work was supported in part by JSPS KAKENHI Grant Number JP19K07243 to M. H
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Masayo Harada designed the study and performed the experiments. Masayo Harada and Keiichi Akita wrote the manuscript.
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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted (Institutional Animal Care and Use Committee of Tokyo Medical and Dental University, permit number A2019-097C). This article does not contain any studies with human participants performed by any of the authors.
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Harada, M., Akita, K. Mouse fibroblast growth factor 9 N143T mutation leads to wide chondrogenic condensation of long bones. Histochem Cell Biol 153, 215–223 (2020). https://doi.org/10.1007/s00418-020-01844-2
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DOI: https://doi.org/10.1007/s00418-020-01844-2