Feeling for Filaments: Quantification of the Cortical Actin Web in Live Vascular Endothelium

24Citations
Citations of this article
52Readers
Mendeley users who have this article in their library.

Abstract

Contact-mode atomic force microscopy (AFM) has been shown to reveal cortical actin structures. Using live endothelial cells, we visualized cortical actin dynamics simultaneously by AFM and confocal fluorescence microscopy. We present a method that quantifies dynamic changes in the mechanical ultrastructure of the cortical actin web. We argue that the commonly used, so-called error signal imaging in AFM allows a qualitative, but not quantitative, analysis of cortical actin dynamics. The approach we used comprises fast force-curve-based topography imaging and subsequent image processing that enhances local height differences. Dynamic changes in the organization of the cytoskeleton network can be observed and quantified by surface roughness calculations and automated morphometrics. Upon treatment with low concentrations of the actin-destabilizing agent cytochalasin D, the cortical cytoskeleton network is thinned out and the average mesh size increases. In contrast, jasplakinolide, a drug that enhances actin polymerization, consolidates the cytoskeleton network and reduces the average mesh area. In conclusion, cortical actin dynamics can be quantified in live cells. To our knowledge, this opens a new pathway for conducting quantitative structure-function analyses of the endothelial actin web just beneath the apical plasma membrane.

Cite

CITATION STYLE

APA

Kronlage, C., Schäfer-Herte, M., Böning, D., Oberleithner, H., & Fels, J. (2015). Feeling for Filaments: Quantification of the Cortical Actin Web in Live Vascular Endothelium. Biophysical Journal, 109(4), 687–698. https://doi.org/10.1016/j.bpj.2015.06.066

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free