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Interference with endogenous ras function inhibits cellular responses to wounding
R G Sosnowski1, S Feldman, J R Feramisco
1Department of Pharmacology, University of California, San Diego, La Jolla 92093-0636.
Abstract:
Wounding of tissue induces cellular responses that ultimately result in wound repair. Studies in tissue culture model systems indicate that these responses include induction of AP-1 regulated genes, cell migration and mitogenesis which are also characteristic of cellular responses to growth factors. Investigations have identified cellular ras proteins as critical components of growth factor-stimulated signal transduction pathways, however their role in the wounding response is less clear. Investigation of the potential involvement of c-Ras in this process utilized quiescent living bovine corneal endothelium cells (BCE) which were microinjected with ras dominant interfering mutant protein (N17) and subsequently stimulated by mechanical wounding. Analysis of these cells demonstrated that microinjection of dominant-interfering ras protein, but not control proteins, inhibited the wounding response as evidenced by diminished Fos expression, lack of cell migration and a block in DNA synthesis.
Insights
Cellular ras proteins are crucial for wound repair. Inhibiting ras signaling in bovine corneal endothelium cells blocked key repair responses like Fos expression and DNA synthesis after wounding.
Area of Science:
- Cellular biology
- Molecular biology
- Tissue repair mechanisms
Background:
- Tissue injury triggers cellular responses essential for repair, including gene induction, migration, and proliferation.
- Cellular ras proteins are known regulators of growth factor signaling, but their role in wound healing is not fully understood.
Purpose of the Study:
- To investigate the involvement of cellular ras proteins in the cellular response to tissue wounding.
- To determine if ras signaling is essential for key wound repair processes in corneal endothelial cells.
Main Methods:
- Utilized quiescent bovine corneal endothelium (BCE) cells in a tissue culture model.
- Microinjected BCE cells with a dominant-interfering ras mutant protein (N17) or control proteins.
- Stimulated cells with mechanical wounding and analyzed cellular responses.
Main Results:
- Microinjection of dominant-interfering ras protein significantly inhibited the wound healing response.
- Inhibition of ras signaling led to diminished Fos expression, a marker of AP-1 regulated genes.
- Ras inhibition also blocked cell migration and DNA synthesis, crucial for tissue repair.
Conclusions:
- Cellular ras proteins play a critical role in mediating the cellular responses to mechanical wounding.
- Ras signaling is essential for the induction of gene expression, cell migration, and proliferation during wound repair.
- Targeting ras pathways may offer therapeutic potential for enhancing wound healing.