Abstract
Adipocyte differentiation is a well defined process that is under the control of transcriptional activators and repressors. We show that histone deacetylase (HDAC) inhibitors efficiently block adipocyte differentiation in vitro. This effect is specific to adipogenesis, as another mesenchymal differentiation process, osteoblastogenesis, is enhanced upon HDAC inhibition. Through the systematic genetic deletion of HDAC genes in cultured mesenchymal precursor cells, we show that deletion of HDAC1 and HDAC2 leads to reduced lipid accumulation, revealing redundant and requisite roles of these class I HDACs in adipogenesis. These findings unveil a previously unrecognized role for HDACs in the control of adipogenesis.
Highlights
In humans, unused caloric energy resulting from an excessive net caloric intake is converted to triglycerides and stored in fat depots for further usage
Through the systematic genetic deletion of histone deacetylase (HDAC) genes in cultured mesenchymal precursor cells, we show that deletion of HDAC1 and HDAC2 leads to reduced lipid accumulation, revealing redundant and requisite roles of these class I HDACs in adipogenesis
This phenotype is not seen in cardiomyocytes deficient for either HDAC1, HDAC2, or HDAC8, indicating that fatty acid and adipocyte homeostasis might be under the control of distinct HDAC isoforms [19, 22]
Summary
Cell Culture and Adipocyte Differentiation—3T3-L1 cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum and antibiotics. D, up-regulation of adipogenic marker genes in induced 3T3-L1 cells is blocked by incubation with TSA, Scriptaid, and SAHA but not sodium butyrate as shown by RT-PCR for different markers of terminally differentiated adipocytes, such as hormone-sensitive lipase (LIPE), adiponectin (AdipoQ), and adipocyte lipid-binding protein (aP2). To detect C/EBP, post-confluent cells were induced to differentiate as described above for 18 h and eventually incubated with 100 nM TSA (Sigma). Cells were fixed, blocked with bovine serum albumin, and incubated with anti-C/EBP primary antibody (mouse monoclonal, 1:200 dilution, Santa Cruz Biotechnology) for 1 h at room temperature. Cells were fixed, treated with 1.5 M HCl, permeabilized, blocked with bovine serum albumin, and incubated with anti-BrdUrd primary antibody (mouse monoclonal, 1:200 dilution, Roche Applied Science) for 1 h at room temperature.
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