Study on bone formation in Ano5-knockout mice
【摘要】：Gnathodiaphyseal dysplasia(GDD; OMIM#166260) is a rare skeletal disorder with autosomal dominant inherit pattern. It is characterized by lesions of jawbones, thickening cortical diaphysis of tubular bones and frequent fractures as a results of minimal injury. We found previously that the silence of Ano5 gene resulted in increased mineral nodule formation in differentiating MC3 T3-E1 osteoblast precursors in vitro and findings suggests that ANO5 plays a role in osteoblast differentiation. However, the pathological effects of ANO5 deficiency on GDD in vivo has not been elucidated completely. Now we generated a Ano5-konckout mouse modal with CRISPR/Cas 9 method. The expression of Ano5 in bone tissue decreased significantly and some clinical features of human GDD have been replicated. Meanwhile, the mouse calvarial osteoblast(mCOBs) cultures were performed and the expression of osteoblast-related genes as well as bone matrix formation assays were investigated by quantitative PCR and alizarin red staining respectively. The results showed that Osteocalcin, Col1 a1, Runx2, Osterix, Osteopontin and Rankl highly elevated and mineralization enhanced drastically in Ano5-/-mCOB. The data are consistent with the achievements we observed before in vitro. We believe this new mouse model can contribute to research into the pathogenesis of skeletal abnormalities in GDD and provide the clues to develop the therapeutic approaches for GDD.