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Related Experiment Videos

Cell density-dependent decrease in cytoskeletal actin and myosin in cultured osteoblastic cells: correlation with

J J Egan1, G Gronowicz, G A Rodan

  • 1Laboratory of Cellular and Developmental Biology, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.

Journal of Cellular Biochemistry
|January 1, 1991
PubMed
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Increased cell density in osteoblasts leads to changes in cell shape and actin-myosin organization. Elevated intracellular cyclic AMP (cAMP) appears to mediate these density-dependent transformations in bone cells.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Bone Biology

Background:

  • Osteoblasts exhibit morphological changes from fibroblast-like to cuboidal with increasing cell density during bone development and in culture.
  • Parathyroid hormone, known to increase intracellular cyclic AMP (cAMP) in osteoblasts, also induces similar shape changes.

Purpose of the Study:

  • To investigate the role of intracellular cyclic AMP (cAMP) in the density-dependent morphological and cytoskeletal changes of osteoblasts.
  • To determine the relationship between cell density, cyclic AMP levels, and the organization of actin and myosin in osteoblast cultures.

Main Methods:

  • Osteoblast-enriched rat calvaria cells were cultured at varying densities.
  • Quantified Triton X-100 extractable and nonextractable actin and myosin using polyacrylamide gel electrophoresis.

Related Experiment Videos

  • Measured intracellular cyclic AMP (cAMP) levels via 3H-adenine prelabeling and HPLC separation.

    • Main Results:

    • Osteoblastic cells contain significant amounts of actin and myosin, with a majority in a nonextractable (crosslinked) form at lower densities.
    • Above 50,000 cells/cm2, cell density correlated with decreased crosslinked actin and myosin and increased intracellular cyclic AMP (cAMP).
    • Agents that increase cyclic AMP (e.g., phosphodiesterase inhibitors) mimicked these changes, while cytochalasin B affected actin polymerization and cell shape.

    Conclusions:

    • Elevated intracellular cyclic AMP (cAMP) may be a key mediator of density-dependent changes in osteoblast cell shape.
    • Cyclic AMP influences microfilament organization and cytoskeletal dynamics in osteoblasts in response to increasing cell density.