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The organization of microfilaments in spreading platelets: a comparison with fibroblasts and glial cells
Abstract:
Platelets respond to stimulatory agents in general by the formation of long spikelike surface projections built up of tightly bundled microfilaments. During contact stimulation this is followed by a second phase when thin membrane lamellae grow out between the projections. This behaviour resembles that seen for instance in fibroblasts and glial cells, sending out microspikes and lamellipodia as a step in their advancement over solid substrata. Conditions, designed earlier for the preservation and visualization of the fragile organization of microfilaments and microtubules in the peripheral, highly motile parts (leading lamellae) of such cells (Höglund et al. (1980) J. Musc. Res. Cell Motility, 1:127-146), were used here to produce high-resolution images of the ultrastructural organization of platelets spreading on a solid substratum. This revealed an unexpected arrangement of actin filaments running parallel to the advancing edge, and small tufts of microfilaments on the outside of this edge-bundle. Cytochalasin D caused a regression of the spikelike projections as well as of both types of structures in the advancing platelet lamella and led to the appearance of a dense filamentous mat in juxtaposition to the plasma membrane. Analysis of the actin pools using the DNAase inhibition assay showed that the dramatic reorganizations of actin seen during the two phases of contact stimulation was reflected in a shift in the G/F-actin ratio only during the early phase.
Insights
Platelets form spiky projections and lamellae when stimulated. This study reveals unexpected actin filament organization during platelet spreading on surfaces, impacting cell motility.
Area of Science:
- Cell Biology
- Biophysics
- Cytoskeleton Dynamics
Background:
- Platelets exhibit dynamic surface projections and lamellae upon stimulation.
- Cellular motility involves microspikes and lamellipodia, similar to platelet behavior.
- Previous methods preserved microfilament and microtubule organization in motile cell regions.
Purpose of the Study:
- To visualize the ultrastructural organization of platelets spreading on a solid substratum.
- To investigate the role of actin filaments in platelet lamellar structures.
- To analyze changes in actin pools during platelet contact stimulation.
Main Methods:
- High-resolution imaging of platelet ultrastructure using preserved cytoskeletal organization.
- Treatment with Cytochalasin D to observe effects on platelet structures.
- DNAase inhibition assay to analyze G/F-actin ratios.
Main Results:
- Revealed an unexpected parallel arrangement of actin filaments along the advancing platelet edge.
- Observed small microfilament tufts outside the edge-bundle.
- Cytochalasin D induced regression of projections and formation of a dense filamentous mat.
- G/F-actin ratio shifts occurred only during the early phase of contact stimulation.
Conclusions:
- Platelet spreading involves a unique actin organization at the leading edge.
- Actin dynamics are crucial for platelet shape changes and motility.
- Cytochalasin D disrupts actin organization, affecting platelet lamellar structures.