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Published on: July 21, 2021
Molecular fingerprinting of BMP2- and BMP4-treated embryonic maxillary mesenchymal cells
P Mukhopadhyay1, S Singh, R M Greene
1Department of Molecular Cellular and Craniofacial Biology, University of Louisville Birth Defects Center, ULSD, University of Louisville, KY 40292, USA.
Objective:
To determine the differences in gene expression between control-, bone morphogenetic protein (BMP)2- and BMP4-treated murine embryonic maxillary mesenchymal (MEMM) cells.
Design:
Transcript profiles of BMP2-, BMP4- and vehicle-treated MEMM cells were compared utilizing the murine high-density GeneChip arrays from Affymetrix. The raw chip data (probe intensities) were pre-processed using robust multichip averaging with GC-content background correction and further normalized with GeneSpring v7.2 software. Cluster analysis of the microarray data was performed with the GeneSpring software. Changes in the gene expression were verified by TaqMan quantitative real-time PCR.
Results:
Expression of approximately 50% of the 45 101 genes and expressed sequence tags examined in this study were detected in BMP2-, BMP4- and vehicle-treated MEMM cells and that of several hundred genes was significantly altered (up or downregulated) in these cells in response to BMP2 and BMP4. Expression profiles of each of the 26 mRNAs tested by TaqMan quantitative real-time PCR were found to be consistent with the microarray data. Genes whose expression was modulated following BMP2 or BMP4 treatment, could be broadly classified based on the functions of the encoded proteins such as the growth factors and signaling molecules, transcription factors, and proteins involved in epithelial-mesenchymal interactions, extracellular matrix synthesis, cell adhesion, proliferation, differentiation, and apoptosis.
Conclusion:
Utilization of the Affymetrix GeneChip microarray technology has enabled us to delineate a detailed transcriptional map of BMP2 and BMP4 responsiveness in embryonic maxillary mesenchymal cells and offers revealing insights into crucial molecular regulatory mechanisms employed by these two growth factors in orchestrating embryonic orofacial cellular responses.
