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Gap junctional communication modulates gene expression in osteoblastic cells

F Lecanda1, D A Towler, K Ziambaras

  • 1Division of Bone and Mineral Diseases, Washington University School of Medicine, and Barnes-Jewish Hospital, St. Louis, Missouri 63110, USA.

Molecular Biology of the Cell
|August 7, 1998
PubMed
Summary
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Altering gap junction communication in bone cells by changing connexin43 (Cx43) and connexin45 (Cx45) expression affects osteoblast gene activity. This suggests a communicating cell network is vital for bone cell development and function.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Biophysics

Background:

  • Bone-forming cells (osteoblasts) form a network connected by gap junctions.
  • These gap junctions are composed of connexin43 (Cx43) and connexin45 (Cx45).
  • Cx43 and Cx45 form gap junctions with differing permeability characteristics.

Purpose of the Study:

  • To investigate how altering gap junctional communication impacts osteoblast phenotype.
  • To determine the role of Cx43 and Cx45 expression levels in osteoblast differentiation.
  • To explore the relationship between gap junction communication and the transcription of bone-specific genes.

Main Methods:

  • Manipulating the expression of Cx43 and Cx45 in osteoblast cell lines (ROS 17/2.8, MC3T3-E1, UMR 106-01) via transfection.

Related Experiment Videos

  • Assessing dye transfer (e.g., Lucifer yellow, calcein) to measure gap junctional communication.
  • Quantifying the expression of osteoblast-specific genes (osteocalcin, bone sialoprotein) using molecular techniques.
  • Analyzing the transcriptional activity of osteoblast promoters using promoter-luciferase constructs.

    • Main Results:

    • Transfecting Cx45 into Cx43-dominant cells decreased dye transfer and expression of osteocalcin (OC) and bone sialoprotein (BSP).
    • Transfecting Cx43 into Cx45-dominant cells increased cell coupling and OC and BSP expression.
    • Cx45 cotransfection down-regulated OC and BSP gene transcription, while Cx43 up-regulated it, correlating with changes in dye coupling.
    • Less specific osteoblast promoters showed reduced sensitivity to altered gap junction communication.

    Conclusions:

    • Altering Cx43 and Cx45 expression in osteoblastic cells modulates the transcriptional activity of osteoblast-specific genes.
    • Changes in gene transcription are likely mediated by signals that diffuse through the gap junction network.
    • A functional intercellular communication network is essential for the complete development of the osteoblastic phenotype.