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Identification and bioinformatic analysis of differentially expressed genes in osteoblasts and osteocytes by genome-wide expression analysis of murine calvarial tissue

To elucidate the differential gene expression between osteoblasts and osteocytes a comprehensive analysis of their transcriptional profiles was completed in this PhD study. Selective identification of these two cell population enzymatically obtained from neonatal murine calvarial tissue was achieved by utilization of dual GFP transgenic mice. Isolation of distinct cell populations was performed by fluorescence activated8 (FACS) cell sorting. Gene expression was analyzed using the Illumina WG-6v1 BeadChip. Altogether, a total of 2541 genes were called present in GFP negative, Col2.3cyan (+) (osteoblasts), and DMP1topaz (+) (preosteocytes and osteocytes) cell populations. The selected genes were bioinformatically classified and grouped according to their associated gene ontology terms. The osteoblastic cell population exhibited high levels of expression h of bone matrix protein genes as well known characteristics of osteoblasts. In contrast, the osteocyte population does not express major matrix protein genes but has relatively high levels of genes (DMP1, Phex and Ank) that participate in the phosphate regulation. A large number of genes associated with structural myofibril components and muscle cell development and function are found highly expressed in osteocytes. The presence of neuropeptide Y (NPY) gene expression, a central regulator of bone mass that exhibit higher levels in DMP1topaz (+) cells was also detected.

osteoblast; osteocyte; transgenic mice; gene expression

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