Journal article
Variation in cell-substratum adhesion in relation to cell cycle phases.
- Meredith DO Interface Biology, AO Research Institute, Davos-Platz, Switzerland.
- Owen GR
- ap Gwynn I
- Richards RG
- 2004-01-20
Published in:
- Experimental cell research. - 2004
3T3 Cells
Animals
Autoradiography
Cell Adhesion
Cell Cycle
Fibroblasts
Focal Adhesions
Genetic Variation
Immunohistochemistry
Mice
Microscopy, Electron, Scanning
S Phase
English
The quantification of focal adhesion sites offers an assessable method of measuring cell-substrate adhesion. Such measurement can be hindered by intra-sample variation that may be cell cycle derived. A combination of autoradiography and immunolabelling techniques, for scanning electron microscopy (SEM), were utilised simultaneously to identify both S-phase cells and their focal adhesion sites. Electron-energy 'sectioning' of the sample, by varying the accelerating voltage of the electron beam, combined with backscattered electron (BSE) imaging, allowed for S-phase cell identification in one energy 'plane' image and quantitation of immunogold label in another. As a result, it was possible simultaneously to identify S-phase cells and their immunogold-labelled focal adhesions sites on the same cell. The focal adhesion densities were calculated both for identified S-phase cells and the remaining non-S-phase cells present. The results indicated that the cell cycle phase was a significant factor in determining the density of focal adhesions, with non-S-phase cells showing a larger adhesion density than S-phase cells. Focal adhesion morphology was also seen to correspond to cell cycle phase; with 'dot' adhesions being more prevalent on smaller non-S-phase and the mature 'dash' type on larger S-phase cells. This study demonstrated that when quantitation of focal adhesion sites is required, it is necessary to consider the influence of cell cycle phases on any data collected.
- Language
-
- English
- Open access status
- closed
- Identifiers
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- DOI 10.1016/j.yexcr.2003.10.005
- PMID 14729057
- Persistent URL
- https://sonar.ch/global/documents/190924
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