Construction of adenovirus vector expressing duck sclerostin and its induction effect on myogenic proliferation and differentiation in vitro.

BACKGROUND

Bones and muscles originated together from the mesoderm during embryogenesis, and they can influence each other through mechanical stimulations and chemical signals. The sclerostin (SOST) is secreted from mature osteocytes. Here, we used a bird model to illustrate the potential roles of SOST on duck myoblasts to verify the hypothesis that SOST might play functions in coordinating the development of bones and muscles.

METHODS AND

RESULTS

Firstly, a recombinant adenovirus vector carrying duck SOST was constructed. Then, the adenovirus-mediated duck SOST was transfected into duck myoblasts. The results revealed by CCK-8 showed that the cell proliferation of myoblasts was inhibited after 12 h, 36 h, and 48 h treatment by transfection of SOST. The labeling rates of EdU positive cells in the Ad-duSOST group were significantly lower than the Ad-NC group (P < 0.05). However, the flow cytometry showed that the cells' G0/G1 phase number was not significantly different. Furthermore, the immunofluorescence results showed that the formation of myotubes was inhibited. Subsequent transcriptome revealed that, under the ectopic expression of SOST, the genes related to Cytokine-cytokine receptor interaction, muscle development (regulation of action cytoskeleton, Wnt signaling pathway), and intercellular regulation were changed. Six of the top 20 DEGs were related to morphogenesis.

CONCLUSIONS

Our studies demonstrated that the SOST played critical roles in myoblasts differentiation by mediating the crosstalk among several pathways and transcription factors related to cell differentiation. Our data provided cellular evidence supporting the combined functions of SOST in coordinating bone and muscle co-development.