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Published on: December 10, 2010
Expression of glutamyl aminopeptidase by osteogenic induction in rat bone marrow stromal cells
Yao Wu1, Jingang Xiao, Ling Wu
1Department of Oral and Maxillofacial Surgery, West China College of Stomatology, Sichuan University, 14 Section 3, Renminnan Road, Chengdu, Sichuan 610041, PR China.
Insights
Glutamyl aminopeptidase (GluAP) expression increases during the osteogenic differentiation of rat bone marrow stromal cells (BMSCs). This suggests GluAP plays a role in bone cell development.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Glutamyl aminopeptidase (GluAP) is a zinc metallopeptidase involved in peptide metabolism.
- Its role in bone cell differentiation is not well understood.
Purpose of the Study:
- To investigate the expression of GluAP during osteogenic differentiation of rat bone marrow stromal cells (BMSCs).
- To explore the potential correlation between GluAP and osteogenesis.
Main Methods:
- Osteogenic induction of rat BMSCs in vitro.
- DNA microarray hybridization (Genechip Rat Expression Array 230A).
- Real-time PCR, MTT assay, alkaline phosphatase assay, alizarin red staining, and immunohistochemistry.
Main Results:
- GluAP expression was significantly upregulated in osteogenic-induced BMSCs compared to non-induced cells.
- Multiple assays confirmed successful osteogenic differentiation of BMSCs.
Conclusions:
- GluAP is upregulated during in vitro osteogenic differentiation of rat BMSCs.
- GluAP may be a novel factor involved in the process of osteogenesis.
Abstract:
Glutamyl aminopeptidase (GluAP, EC 3.4.11.7, ENPEP) is a 130-kDa homodimeric zinc metallopeptidase which specifically cleaves the N-terminal glutamate or aspartate residue of peptidic substrates such as cholecystokinin-8 or angiotensin (Ang) II, in vitro. We used a DNA microarray hybridization (Genechip Rat Expression Array 230A, Affymetrix Inc., Santa Clara, CA, USA) to demonstrate that GluAP was upregulated in osteogenic induced rat bone marrow stromal cells (BMSCs). To compare the expression of GluAP in the osteogenic differentiation and non-osteogenic differentiation of rat BMSCs in vitro, the cells were osteogenic induced in vitro. We also performed an MTT assay, alkaline phosphatase assay, alizarin red staining, and an immunohistochemical analysis to determine the osteogenic differentiation of BMSCs. The expression of GluAP was examined by real-time polymerase chain reaction (PCR). The real-time PCR results showed that GluAP was upregulated in osteogenic differentiated BMSCs in vitro, suggesting that GluAP may be correlated with the osteogenic differentiation of BMSCs.
