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Vol. 9, Issue 8, 2249-2258, August 1998
§§ and§
*Division of Bone and Mineral Diseases, Departments of
Bone-forming cells are organized in a multicellular network
interconnected by gap junctions. In these cells, gap junctions are
formed by connexin43 (Cx43) and connexin45 (Cx45). Cx43 gap junctions
form pores that are more permeable to negatively charged dyes such as
Lucifer yellow and calcein than are Cx45 pores. We studied whether
altering gap junctional communication by manipulating the relative
expression of Cx43 and Cx45 affects the osteoblast phenotype.
Transfection of Cx45 in cells that express primarily Cx43 (ROS 17/2.8
and MC3T3-E1) decreased both dye transfer and expression of osteocalcin
(OC) and bone sialoprotein (BSP), genes pivotal to bone matrix
formation and calcification. Conversely, transfection of Cx43 into
cells that express predominantly Cx45 (UMR 106-01) increased both cell
coupling and expression of OC and BSP. Transient cotransfection of
promoter-luciferase constructs and connexin expression vectors
demonstrated that OC and BSP gene transcription was down-regulated by
Cx45 cotransfection in ROS 17/2.8 and MC3T3-E1 cells, in association
with a decrease in dye coupling. Conversely, cotransfection of Cx43 in
UMR 106-01 cells up-regulated OC and BSP gene transcription. Activity
of other less specific osteoblast promoters, such as osteopontin and
osteonectin, was less sensitive to changes in gap junctional
communication. Thus, altering gap junctional permeability by
manipulating the expression of Cx43 and Cx45 in osteoblastic cells
alters transcriptional activity of osteoblast-specific promoters,
presumably via modulation of signals that can diffuse from cell to
cell. A communicating intercellular network is required for the full
elaboration of a differentiated osteoblastic phenotype.
Medicine,
Molecular Biology,
Molecular Biology of the Cell
Vol. 9, 2249-2258, August 1998
Copyright © 1998 by The American Society for Cell Biology
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