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Article · Scientific Reports · August 21, 2026

High temporal resolution tracking reveals region-specific control of focal adhesion nucleation, dynamics, and translocation

Vadim V. Mustyatsa, Arina Zhuravel, Iliyas Marat, Nurdaulet Kulsharip, Madina Tlegenova, Aleena Saidova

Ivan Vorobjev
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Abstract

Focal adhesions (FAs) are dynamic multiprotein complexes linking actin cytoskeleton to extracellular matrix and involved in cell attachment, spreading and motility. However, quantitative population analysis of FAs remains limited, as previous studies have primarily focused on selected FA subpopulations, leaving the full progression and coordination of FA behaviors across the entire cell underexplored. To address this, we developed an automated tracking framework that reconstructs complete FA life histories with high spatial and temporal resolution, simultaneously quantifying numerous FA parameters from formation to disassembly. Applying this approach to A549 cells revealed that nascent and mature adhesions constitute a continuous dynamic population dominated by small, short-lived structures. Multiparametric automated analysis uncovered several novel FA characteristics: distinct assembly and disassembly kinetics for FAs of similar size, stable protein density across a wide range of FA sizes, intermittent upregulation of FA formation behind rapidly advancing lamellae, coordinated paxillin and vinculin dynamics and pronounced FA translocation in retracting areas, accompanied by instances of separation of paxillin- and vinculin-containing FA layers. Microtubule depolymerization stabilized FAs and slowed turnover kinetics while preserving region-specific adhesion dynamics. We suggest microtubules play a dual role in FA dynamics, stimulating formation of FAs behind advancing lamellipodia and accelerating turnover of FAs in the entire cell. Overall, our framework enables comprehensive automated analysis of FA behavior across multiple proteins and dynamic states from the earliest stages of formation.

Cellular Mechanics and InteractionsCell Adhesion Molecules ResearchCell Image Analysis Techniquesfocal adhesionsfocal adhesion dynamicsfocal adhesion turnover

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