The Osteoblast Transcriptome in Developing Zebrafish Reveals Key Roles for Extracellular Matrix Proteins Col10a1a and Fbln1 in Skeletal Development and Homeostasis
Ratish Raman,
Mishal Antony,
Renaud Nivelle,
Arnaud Lavergne,
Jérémie Zappia,
Gustavo Guerrero-Limón,
Caroline Caetano da Silva,
Priyanka Kumari,
Jerry Maria Sojan,
Christian Degueldre,
Mohamed Ali Bahri,
Agnes Ostertag,
Corinne Collet,
Martine Cohen-Solal,
Alain Plenevaux,
Yves Henrotin,
Jörg Renn,
Marc Muller
Affiliations
Ratish Raman
Laboratory for Organogenesis and Regeneration (LOR), GIGA Institute, University of Liège, 4000 Liège, Belgium
Mishal Antony
Laboratory for Organogenesis and Regeneration (LOR), GIGA Institute, University of Liège, 4000 Liège, Belgium
Renaud Nivelle
Laboratory for Organogenesis and Regeneration (LOR), GIGA Institute, University of Liège, 4000 Liège, Belgium
Arnaud Lavergne
GIGA Genomics Platform, B34, GIGA Institute, University of Liège, 4000 Liège, Belgium
Jérémie Zappia
MusculoSKeletal Innovative Research Lab, Center for Interdisciplinary Research on Medicines, University of Liège, 4000 Liège, Belgium
Gustavo Guerrero-Limón
Laboratory for Organogenesis and Regeneration (LOR), GIGA Institute, University of Liège, 4000 Liège, Belgium
Caroline Caetano da Silva
Hospital Lariboisière, Reference Centre for Rare Bone Diseases, INSERM U1132, Université de Paris-Cité, F-75010 Paris, France
Priyanka Kumari
Laboratory of Pharmaceutical and Analytical Chemistry, Department of Pharmacy, CIRM, Sart Tilman, 4000 Liège, Belgium
Jerry Maria Sojan
Department of Life and Environmental Sciences, Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, Italy
Christian Degueldre
GIGA CRC In Vivo Imaging, University of Liège, Sart Tilman, 4000 Liège, Belgium
Mohamed Ali Bahri
GIGA CRC In Vivo Imaging, University of Liège, Sart Tilman, 4000 Liège, Belgium
Agnes Ostertag
Hospital Lariboisière, Reference Centre for Rare Bone Diseases, INSERM U1132, Université de Paris-Cité, F-75010 Paris, France
Corinne Collet
Hospital Lariboisière, Reference Centre for Rare Bone Diseases, INSERM U1132, Université de Paris-Cité, F-75010 Paris, France
Martine Cohen-Solal
Hospital Lariboisière, Reference Centre for Rare Bone Diseases, INSERM U1132, Université de Paris-Cité, F-75010 Paris, France
Alain Plenevaux
GIGA CRC In Vivo Imaging, University of Liège, Sart Tilman, 4000 Liège, Belgium
Yves Henrotin
MusculoSKeletal Innovative Research Lab, Center for Interdisciplinary Research on Medicines, University of Liège, 4000 Liège, Belgium
Jörg Renn
Laboratory for Organogenesis and Regeneration (LOR), GIGA Institute, University of Liège, 4000 Liège, Belgium
Marc Muller
Laboratory for Organogenesis and Regeneration (LOR), GIGA Institute, University of Liège, 4000 Liège, Belgium
Zebrafish are now widely used to study skeletal development and bone-related diseases. To that end, understanding osteoblast differentiation and function, the expression of essential transcription factors, signaling molecules, and extracellular matrix proteins is crucial. We isolated Sp7-expressing osteoblasts from 4-day-old larvae using a fluorescent reporter. We identified two distinct subpopulations and characterized their specific transcriptome as well as their structural, regulatory, and signaling profile. Based on their differential expression in these subpopulations, we generated mutants for the extracellular matrix protein genes col10a1a and fbln1 to study their functions. The col10a1a−/− mutant larvae display reduced chondrocranium size and decreased bone mineralization, while in adults a reduced vertebral thickness and tissue mineral density, and fusion of the caudal fin vertebrae were observed. In contrast, fbln1−/− mutants showed an increased mineralization of cranial elements and a reduced ceratohyal angle in larvae, while in adults a significantly increased vertebral centra thickness, length, volume, surface area, and tissue mineral density was observed. In addition, absence of the opercle specifically on the right side was observed. Transcriptomic analysis reveals up-regulation of genes involved in collagen biosynthesis and down-regulation of Fgf8 signaling in fbln1−/− mutants. Taken together, our results highlight the importance of bone extracellular matrix protein genes col10a1a and fbln1 in skeletal development and homeostasis.
