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Abstract
Bone morphogenetic proteins (BMPs) are important regulators of cellular differentiation and embryonic development. Beta catenin mediated nuclear signaling has been implicated in BMP-2-modulated chondrogenic differentiation in the pluripotential stem cell line C3H10T1/2. However, there is little information on the functional role of beta catenin in BMP-2-modulated differentiation of primary nontransformed mesenchymal cells. Here, we present evidence to show that BMP-2-induced chondrogenic differentiation in high-density primary mesenchymal culture is associated with a significant decrease in membrane-bound beta catenin by 72 hours compared to controls. Nuclear localization of beta catenin is not detectable by immunofluorescence and the TCF-responsive reporter vector TOPFLASH shows only background activity during chondrogenic differentiation. BMP-2-treated cultures show reduced cell-cell adhesion by 72 hours, which correlates with the changes in levels of membrane-bound beta catenin. Upregulation of membrane-bound beta catenin blocks the effect of BMP-2 on both chondrogenic differentiation and cell-cell adhesiveness. These findings suggest that BMP-2 can modulate the adhesivity of adherens junctions through regulation of membrane bound beta catenin.
| Abbreviations | ||
| BCA | = | bicinchoninic acid |
| BMP | = | bone morphogenetic protein |
| BSA | = | bovine serum albumin |
| COX-2 | = | cyclo-oxygenase-2 |
| DAPI, | = | 4′,6-diamidino-2-phenylindole dihydrochloride |
| ECL | = | electrochemiluminescent |
| EDTA | = | sodium ethylenediamine tetraacetate |
| FCS | = | fetal calf serum |
| GSK3beta | = | glycogen synthase kinase 3beta |
| HBSS | = | Hanks buffered saline solution |
| HCL | = | hydrochloric acid |
| HMG | = | high mobility group |
| HRP | = | horseradish peroxidase |
| LEF | = | lymphoid enhancer binding factor |
| PBS | = | phosphate buffered saline |
| PCR | = | polymerase chain reaction |
| R.T. | = | room temperature |
| SDS | = | sodium dodecyl sulphate |
| TCF | = | T-cell transcription factor |
| TGF-beta | = | transforming growth factor beta |
| Abbreviations | ||
| BCA | = | bicinchoninic acid |
| BMP | = | bone morphogenetic protein |
| BSA | = | bovine serum albumin |
| COX-2 | = | cyclo-oxygenase-2 |
| DAPI, | = | 4′,6-diamidino-2-phenylindole dihydrochloride |
| ECL | = | electrochemiluminescent |
| EDTA | = | sodium ethylenediamine tetraacetate |
| FCS | = | fetal calf serum |
| GSK3beta | = | glycogen synthase kinase 3beta |
| HBSS | = | Hanks buffered saline solution |
| HCL | = | hydrochloric acid |
| HMG | = | high mobility group |
| HRP | = | horseradish peroxidase |
| LEF | = | lymphoid enhancer binding factor |
| PBS | = | phosphate buffered saline |
| PCR | = | polymerase chain reaction |
| R.T. | = | room temperature |
| SDS | = | sodium dodecyl sulphate |
| TCF | = | T-cell transcription factor |
| TGF-beta | = | transforming growth factor beta |