Project description:Recently, serotonin and serotonin reuptake inhibitor (SSRI) drugs have been shown to have an effect on the development and maintenance of bone. However, little is known about its role in craniofacial development. We used microarray as one tool to determine the gene expression profile of intramembranous (calvarial) derived murine pre-osteoblasts after citalopram (SSRI) exposure.
Project description:Recently, serotonin and serotonin reuptake inhibitor (SSRI) drugs have been shown to have an effect on the development and maintenance of bone. However, little is known about its role in craniofacial development. We used microarray as one tool to determine the gene expression profile of intramembranous (calvarial) derived murine pre-osteoblasts after citalopram (SSRI) exposure. We isolated whole RNA from MC3T3-E1 cells treated with proliferation media or proliferation media with SSRI at a dose of 10^-4 mol./liter for 3 or 7 days in culture. We then used an Affymetrix array and compared expression profiles between control and experimental treatments.
Project description:Effects of Ilex paraguariensis and Ligularia fischeri extracts on osteogenesis and bone microenvironment in mouse osteoblasts (MC3T3-E1)
Project description:Effects of Ilex paraguariensis and Ligularia fischeri extracts on osteogenesis and bone microenvironment in mouse osteoblasts (MC3T3-E1)
Project description:Background: It has widely been observed that young children are capable of reossifying large calvarial defects, while adults lack this endogenous tissue-engineering capacity. The ability of juvenile animals to regenerate calvarial defects has been investigated in multiple animal models, including mice. In this study, the authors used cDNA microarrays to investigate the expression of osteogenesis-associated genes upstream and downstream of Runx2 in juvenile and adult mouse calvaria. Methods: Nonsuture-associated parietal bone discs were harvested from 6-day-old (n = 50) and 60-day-old (n = 35) male CD-1 mice. After separation of the underlying dura mater and overlying pericranium, the calvarial discs were snap-frozen and RNA was extracted from pooled samples of calvaria for microarray analysis. Genes analyzed included cytokines, receptors, and cell-surface and matrix proteins both upstream and downstream of Runx2. Results: Genes associated with the Runx2 pathway had notably higher levels in the juvenile versus adult calvaria. All genes except for osteocalcin were expressed at least twofold higher in the juvenile calvaria. This pattern was validated with quantitative real-time polymerase chain reaction. In addition, mRNA for potent osteoinductive growth factors was present at higher levels in the juvenile compared with the adult calvaria. Conclusions: These findings reflect a genomic environment of active osteoblast differentia-tion and ossification in the juvenile calvaria compared with the adult aquiescent calvarial tissue. These data suggest that a decreased osteogenic potential of adult calvarial osteoblasts may, in part, explain the inability of adult animals to heal calvarial defects.
Project description:Sp7/Osterix is a master regulator of osteoblast specification. To identify the Sp7-mediated gene regulatory network in osteoblasts, we performed Sp7 ChIP-seq on primary mouse calvarial osteoblasts comparing the DNA binding profile with the transcriptional profile of Sp7-positive osteoblasts. Analysis of these identified a network of Sp7 regulated osteoblast targets and provides a new insight into the mode of Sp7 action in osteoblast. To further study for the Sp7 mode, we performed ChIP-seq for Sp1, Sp7, Dlx5, as a potential Sp7 partner identified in this study and mutated Sp7 which has mutations in the zinc finger domain, by using in vitro system with a pre-osteoblast mouse cell line, MC3T3E1.
