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Peneti, S. K.

Publications and source records attributed to Peneti, S. K..

2 recordsLinked to original sources

Vimentin bridges scales to convert polarized cell locomotion into coordinated collective migration

Collective cell migration is central in development and disease. Vimentin is an intermediate filament protein expressed by epithelial cells at the edge of wounds where collective cell migration is most efficient. Yet, its functional role in this context remains underexplored. Here, we show that vimentin, over-expressed in cells undergoing partial epithelial to mesenchymal transition at the edge of epithelial monolayers, has a multiscale impact on the whole monolayer mechano-dynamics. Vimentin knock-down delays wound closure, reduces cell coordination, while increasing traction forces exerted by cells on the substratum. It also disrupts the directionality of leader cells migration, as well as the cohesion and coordinated motion of cells deep in the monolayer. We further show that vimentin promotes the conversion of polarized cell locomotion into coordinate collective migration by polarizing actin, focal adhesions and traction forces, sustaining leader cells lamellipodium protrusive activity and directionality, while allowing mechanical coupling of leader with follower cells. Altogether, we show that vimentin is essential for bridging polarized single cell locomotion and coordinated collective migration to allow efficient collective migration. Significant statementEpithelial cells migrate coordinately to repair tissue. Vimentin, transiently enriched at the wound edge plays a key role in guiding this process by converting polarized cell locomotion into coordinate collective migration, enabling leader cells to polarize, form stable protrusions, and maintain directed migration, while allowing mechanical coupling of front cells with followers, necessary for efficient coordinated collective migration. These findings reveal vimentin, a known EMT marker, as a novel positive regulator of efficient wound healing.

cell biology↗

Nuclear pores as versatile reference standards for quantitative superresolution microscopy

Quantitative fluorescence and superresolution microscopy are often limited by insufficient data quality or artifacts. In this context, it is essential to have biologically relevant control samples to benchmark and optimize the quality of microscopes, labels and imaging conditions.\n\nHere we exploit the stereotypic arrangement of proteins in the nuclear pore complex as in situ reference structures to characterize the performance of a variety of microscopy modalities. We created four genome edited cell lines in which we endogenously labeled the nucleoporin Nup96 with mEGFP, SNAP-tag or HaloTag or the photoconvertible fluorescent protein mMaple. We demonstrate their use a) as 3D resolution standards for calibration and quality control, b) to quantify absolute labeling efficiencies and c) as precise reference standards for molecular counting.\n\nThese cell lines will enable the broad community to assess the quality of their microscopes and labels, and to perform quantitative, absolute measurements.

biophysics↗