Function of new candidate genes in zebrafish bone formation. Relevance for human (and general) skeletal pathology.

ESR23 Zebrafish models have been proven relevant to study skeletal pathologies encountered in humans such as osteoporosis or scoliosis but possibly also in farmed fish ARS has identified the FIBULIN3 protein to be upregulated in serum of osteoarthritic patients UPD7 has identified several variants of genes in cohorts of young patients with osteoporosis osteoarthritis or ectopic mineral deposits in the cartilage such as chondrocalcinosis The function of these factors will be evaluated at the wholebody level by generating the zebrafish mutants in the genes of interest eg fbln1 fbln3 il11ra lrp5 wnt1 iftim5 plekhm1 using the CRISPR methodologies The mutant fish phenotype is characterized using established skeletal staining methods fluorescent transgenic lines microCT histomorphometric methods and by determining the expression of marker genes by in situ hybridization or RTPCR Variant proteins will be tested for proliferation in human and fish osteoblast cell culture Genes affecting skeletal development will be characterized in aquaculture species and their expression will be followed in challenging conditions that generate skeletal malformations including exposure to altered gravity conditions to evaluate their contribution to the defects