Abstract
The actin cytoskeleton with its dynamic properties serves as the driving force for the movement and division of cells and gives the cell shape and structure. Disorders in the actin cytoskeleton occur in many diseases. Deeper understanding of its regulation is essential in order to better understand these biochemical processes. In our study, we use metal-induced energy transfer (MIET) as a tool to quantitatively examine the rarely considered third dimension of the actin cytoskeleton with nanometer accuracy. In particular, we investigate the influence of different drugs acting on the ROCK pathway on the three-dimensional actin organization. We find that cells treated with inhibitors have a lower actin height to the substrate while treatment with a stimulator for the ROCK pathway increases the actin height to the substrate, while the height of the membrane remains unchanged. This reveals the precise tuning of adhesion and cytoskeleton tension, which leads to a rich three-dimensional structural behaviour of the actin cytoskeleton. This finetuning is differentially affected by either inhibition or stimulation. The high axial resolution shows the importance of the precise finetuning of the actin cytoskeleton and the disturbed regulation of the ROCK pathway has a significant impact on the actin behavior in the z dimension.
Highlights
The cells cytoskeleton gives mechanical support, allows the cell to deliver cargo and is fundamental for cell division
We investigate the regulation of the actin cytoskeleton through different targeting sites in the Rho-associated coiled coil-containing protein kinase (ROCK) pathway
The major goal of this study was the examination of the distance from the substrate to the actin cytoskeleton, a parameter we call “actin height”
Summary
The cells cytoskeleton gives mechanical support, allows the cell to deliver cargo and is fundamental for cell division. It consists of three biological polymers: microtubules, intermediate filaments and actin. Actin is located in a variety of areas of the cell (Figure 1a). It is found in the cell cortex to give the cell strength and shape [1]. To divide cells, it forms the contractile ring between the nuclei [2,3]. It forms lamellipodia and filopodia [4,5,6,7]
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