Enhancement of Connexin 43 Expression Increases Proliferation and Differentiation of an Osteoblast-like Cell Line

• Published in: Experimental cell research 2001-04, Vol.264 (2), p.397-407
• Main Authors: Gramsch, B., Gabriel, H.-D., Wiemann, M., Grümmer, R., Winterhager, E., Bingmann, D., Schirrmacher, K.
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - metabolism; Animals; Blotting, Northern - methods; Cell Adhesion - physiology; Cell Differentiation; Cell Division; Connexin 43 - biosynthesis; Connexin 43 - genetics; differentiation; electric coupling; Electrophysiology; gap junctions; Gap Junctions - physiology; Gene Expression; osteoblast-like cells; Osteoblasts - cytology; proliferation; Rats; Transfection; Tumor Cells, Cultured
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1006/excr.2000.5145

Bone cells form a functional syncytium as they are coupled by gap junctions composed mainly of connexin 43 (Cx43). To further understand the role of Cx43 in bone cell growth and differentiation, we stably transfected Cx45-expressing UMR 106-01 cells with Cx43 using an expression vector containing rat Cx43 cDNA. Three stably transfected clones were analyzed, all of which showed altered expression of Cx43 and/or Cx45 as was obvious from immunocytochemistry and Northern blotting. Double whole-cell patch clamping revealed single-channel conductances of 20 (Cx45) and 60 pS (Cx43). The overexpression of Cx43 led to an increase in dye coupling concomitant with elevated gap-junctional conductance. The phenotype of the transfected clones was characterized by an increased proliferation (4- to 7-fold) compared to controls. Moreover, a transfectant clone with 10- to 12-fold enhanced Cx43 expression showed a significantly increased calcium content of the extracellular matrix and enlarged mineralization nodules, while alkaline phosphatase was moderately increased. We conclude that enhanced gap-junctional coupling via Cx43 significantly promotes proliferation and differentiation of UMR cells.