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Actin polymerization and intracellular solvent flow in cell surface blebbing

C C Cunningham1

  • 1Department of Medicine, Brigham & Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.

The Journal of Cell Biology
|June 1, 1995
PubMed
Summary

This study explores how cell surface blebs form and expand. Blebs are balloon-like protrusions that appear when cells spread. The researchers found that bleb growth is driven by fluid flow and limited by actin polymerization. They observed that blebs expand rapidly when actin concentration is low. As actin structures strengthen, bleb size and frequency decrease. The study also showed that the protein ABP-280 enhances this actin stabilization. The findings suggest that fluid-driven expansion precedes actin formation in bleb dynamics. This process may also influence other cell protrusions like ruffles.

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Area of Science:

  • Cell membrane dynamics in biophysics
  • Actin cytoskeleton regulation in cell biology
  • Cancer cell motility in oncology

Background:

The regulation of cell surface protrusions remains poorly understood. While actin structures are known to influence membrane behavior, the role of fluid flow in this process is unclear. Established knowledge shows that actin cross-linking proteins like ABP-280 stabilize the cortical actin network. However, how this affects bleb formation is unknown. This gap motivated the investigation of bleb dynamics in melanoma cells. Prior research has shown that blebs alternate with ruffles, suggesting shared mechanisms. Yet, the sequence of events in bleb formation is not fully resolved. This paper's contribution lies in linking bleb expansion to actin polymerization rates. The study addresses whether fluid flow alone can initiate bleb formation. It also examines how ABP-280 influences this process.

Purpose Of The Study:

This study aimed to clarify the relationship between actin polymerization and bleb dynamics in human melanoma cells. The specific problem addressed is whether bleb expansion is driven by fluid flow and limited by actin gelation. The motivation arises from the observation that blebs occur in cells with altered actin cross-linkers. The researchers sought to determine if bleb size correlates with actin concentration. They also wanted to test if ABP-280 affects bleb growth rates. The study focused on how rapidly expanding blebs are halted by actin structures. The goal was to determine if fluid-driven expansion precedes actin polymerization. This could clarify how membrane protrusions are regulated in cancer cells.

Keywords:
actin dynamicscell membrane expansioncancer cell motilitycytoskeletal regulation

Frequently Asked Questions

The authors propose that fluid flow drives bleb expansion before actin polymerization occurs.

ABP-280 expressing cells show decreased bleb growth rates at lower actin concentrations.

Blebs lack initial actin structures, which appear later to stop expansion.

Higher F-actin concentrations reduce bleb expansion rates and final size.

Fluorescent actin labeling and phalloidin injections tracked polymerization events.

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Main Methods:

The researchers used human melanoma cell lines with and without ABP-280 expression. They observed bleb dynamics during cell spreading using fluorescence microscopy. Fluorescently labeled actin was used to track polymerization events. Phalloidin injections helped identify F-actin structures in blebbing cells. The study measured bleb expansion rates and correlated them with actin concentration. Time-lapse imaging captured bleb formation and cessation. The cells were plated on surfaces to monitor gelation over time. The absence of initial actin structures in blebs was confirmed through these methods.

Main Results:

Bleb expansion rates varied and predicted final bleb size. These rates decreased as F-actin concentration increased. In ABP-280 expressing cells, this decrease occurred at lower actin concentrations. Fluorescent labeling showed no initial actin structures in blebs. Polymerized actin structures appeared later and halted bleb expansion. The study found that fluid flow drives bleb expansion before actin gelation. As the actin network strengthens, bleb size and frequency decrease. This suggests that fluid-driven expansion is rate-limited by actin polymerization.

Conclusions:

The authors propose that bleb formation occurs when fluid flow outpaces actin polymerization. They suggest that actin gelation limits bleb expansion regardless of ABP-280 presence. The findings imply that fluid-driven expansion is a primary mechanism in bleb formation. The study supports the idea that actin concentration modulates bleb size. The results suggest that ABP-280 enhances actin gelation efficiency. The researchers propose that this mechanism may influence other protrusions like ruffles. They conclude that bleb dynamics are regulated by the interplay of fluid flow and actin polymerization. These findings may inform future studies on membrane protrusion control.

The authors suggest this mechanism may influence other protrusions like ruffles.