Search

Background: Understanding fate choice and fate switching between the osteoblast lineage (ObL) and adipocyte lineage (AdL) is important to understand both the developmental inter-relationships between osteoblasts and adipocytes and the impact of changes in fate allocation between the two lineages in normal aging and certain diseases. The goal of this study was to determine when during lineage progression ObL cells are susceptible to an AdL fate switch by activation of endogenous peroxisome proliferator-activated receptor (PPAR)c. Methodology/Principal Findings: Multiple rat calvaria cells within the ObL developmental hierarchy were isolated by either fractionation on the basis of expression of alkaline phosphatase or retrospective identification of single cell-derived colonies, and treated with BRL-49653 (BRL), a synthetic ligand for PPARc. About 30% of the total single cell-derived colonies expressed adipogenic potential (defined cytochemically) when BRL was present. Profiling of ObL and AdL markers by qRTPCR on amplified cRNA from over 160 colonies revealed that BRL-dependent adipogenic potential correlated with endogenous PPARc mRNA levels. Unexpectedly, a significant subset of relatively mature ObL cells exhibited osteoadipogenic bipotentiality. Western blotting and immunocytochemistry confirmed that ObL cells co-expressed multiple mesenchymal lineage determinants (runt-related transcription factor 2 (Runx2), PPARc, Sox9 and MyoD which localized in the cytoplasm initially, and only Runx2 translocated to the nucleus during ObL progression. Notably, however, some cells exhibited both PPARc and Runx2 nuclear labeling with concomitant upregulation of expression of their target genes with BRL treatment. Conclusions/Significance: We conclude that not only immature but a subset of relatively mature ObL cells characterized by relatively high levels of endogenous PPARc expression can be switched to the AdL. The fact that some ObL cells maintain capacity for adipogenic fate selection even at relatively mature developmental stages implies an unexpected plasticity with important implications in normal and pathological bone development., This work was supported by Grants-in-Aid from the Ministry of Education, Science, Sports and Culture of Japan (13771074 to YY) and the Canadian Institutes of Health Research (FRN 83704 to JEA). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.