Characterize osteocyte transcriptome under control of PPARa

RNA-seq analysis of transcriptome of osteocytes isolated from femora cortical bone of 5 mo old C57BL6 mice with WT or PPARa KO genotype. Conclusion: In osteocytes, PPARa controls large number of transcripts coding for signaling proteins and osteocyte secretome which regulates development of bone marrow adipose tissue and peripheral fat metabolism. Overall design: Cell fraction enriched in osteocytes was isolated by sequential collagenase digestion of femora bone according to previously described protocol. In brief, isolated femurs were cleaned and scrapped to remove soft tissues and periosteum, respectively. Bone marrow was spun down and epiphysis were removed. Bone diaphysis were sequentially digested in buffers supplemented with collagenase or EDTA. Fraction number 3 which is an osteoblastic fraction and fraction number 6, osteocytic fractions were lysed with Tri Reagent and stored at -80°C. Total RNA was extracted from the samples according to the manufacturer's protocol.