Direct visualization and quantitative analysis of epidermal growth factor-induced receptor clustering
Article first published online: 4 FEB 2005
Copyright © 1988 Wiley-Liss, Inc.
Journal of Cellular Physiology
Volume 134, Issue 3, pages 413–420, March 1988
How to Cite
van Belzen, N., Rijken, P. J., Hage, W. J., de Laat, S. W., Verkleij, A. J. and Boonstra, J. (1988), Direct visualization and quantitative analysis of epidermal growth factor-induced receptor clustering. J. Cell. Physiol., 134: 413–420. doi: 10.1002/jcp.1041340312
- Issue published online: 4 FEB 2005
- Article first published online: 4 FEB 2005
- Manuscript Accepted: 30 OCT 1987
- Manuscript Received: 19 AUG 1987
Several observations have indicated that clustering of growth factor receptors plays an important role in the action of growth factors. In this investigation, we have used the label fracture method to study the effects of epidermal growth factor (EGF) on the lateral distribution of its receptors in A431 epidermoid carcinoma cells. This method allows a direct visualization of immunogold-labeled plasma membrane receptors on ultrastructural level and in addition permits an quantitative analysis of their lateral distribution. EGF receptors were immunogold-labeled according to standard procedures with the monoclonal anti-EGF receptor antibody 2E9 (lgG1), which binds to the EGF receptor in a 1:1 ratio. In the absence of EGF, EGF receptors located on the surface of A431 cells were found to be clustered, as deduced from Poisson variance analysis (p < 0.001). Following treatment of A431 cells with EGF, receptor clustering increased rapidly, reaching the maximum within 10 min. Maximal clustering was maintained for 1 h, after which the lateral distribution of receptors returned to the control situation within another hour.