Microfabricated tissues for investigating traction forces involved in cell migration and tissue morphogenesis

Nerger, Bryan A.; Siedlik, Michael J.; Nelson, Celeste M.

doi:10.1007/s00018-016-2439-z

Cited by 26 publications

(22 citation statements)

References 163 publications

(252 reference statements)

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“…Forces not only generate form and flows within tissues [3,4] but can also control cell fate decisions [5,6] and trigger mitosis [7]. There are various ways to quantify forces at the cellular or tissue level [8,9]; however, mechanical forces experienced by proteins in cells have only recently become quantifiable with the development of Förster resonance energy transfer (FRET)-based molecular tension sensors [10]. These sensors contain a donor and an acceptor fluorophore connected by a mechanosensitive linker peptide, which reversibly unfolds and extends when experiencing mechanical forces.…”

Section: Introductionmentioning

confidence: 99%

A small proportion of Talin molecules transmit forces at developing muscle attachments in vivo

et al. 2019

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Cells in developing organisms are subjected to particular mechanical forces that shape tissues and instruct cell fate decisions. How these forces are sensed and transmitted at the molecular level is therefore an important question, one that has mainly been investigated in cultured cells in vitro. Here, we elucidate how mechanical forces are transmitted in an intact organism. We studied Drosophila muscle attachment sites, which experience high mechanical forces during development and require integrin-mediated adhesion for stable attachment to tendons. Therefore, we quantified molecular forces across the essential integrin-binding protein Talin, which links integrin to the actin cytoskeleton. Generating flies expressing 3 Förster resonance energy transfer (FRET)-based Talin tension sensors reporting different force levels between 1 and 11 piconewton (pN) enabled us to quantify physiologically relevant molecular forces. By measuring primary Drosophila muscle cells, we demonstrate that Drosophila Talin experiences mechanical forces in cell culture that are similar to those previously reported for Talin in mammalian cell lines. However, in vivo force measurements at developing flight muscle attachment sites revealed that average forces across Talin are comparatively low and decrease even further while attachments mature and tissue-level tension remains high. Concomitantly, the Talin concentration at attachment sites increases 5-fold as quantified by fluorescence correlation spectroscopy (FCS), suggesting that only a small proportion of Talin molecules are mechanically engaged at any given time. Reducing Talin levels at late stages of muscle development results in muscle–tendon rupture in the adult fly, likely as a result of active muscle contractions. We therefore propose that a large pool of adhesion molecules is required to share high tissue forces. As a result, less than 15% of the molecules experience detectable forces at developing muscle attachment sites at the same time. Our findings define an important new concept of how cells can adapt to changes in tissue mechanics to prevent mechanical failure in vivo.

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Section: Introductionmentioning

confidence: 99%

A small proportion of Talin molecules transmit forces at developing muscle attachments in vivo

et al. 2019

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confidence: 99%

Traction forces control cell-edge dynamics and mediate distance-sensitivity during cell polarization

Messi

Bornert

Raynaud

et al. 2019

Preprint

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Traction forces are generated by cellular actin-myosin system and transmitted to the environment through adhesions. They are believed to drive cell motion, shape changes, and extracellular matrix remodeling [1][2][3]. However, most of the traction force analysis has been performed on stationary cells, investigating forces at the level of individual focal adhesions or linking them to static cell parameters such as area and edge curvature [4][5][6][7][8][9][10]. It is not well understood how traction forces are related to shape changes and motion, e.g. forces were reported to either increase or drop prior to cell retraction [11][12][13][14][15]. Here, we analyze the dynamics of traction forces during the protrusion-retraction cycle of polarizing fish epidermal keratocytes and find that forces fluctuate in concert with the cycle, increasing during the protrusion phase and reaching maximum at the beginning of retraction. We relate force dynamics to the recently discovered phenomenological rule [16] that governs cell edge behavior during keratocyte polarization: both traction forces and the probability of switch from protrusion to retraction increase with the distance from the cell center. Diminishing traction forces with cell contractility inhibitor leads to decreased edge fluctuations and abnormal polarization, while externally applied force can induce protrusion-retraction switch. These results suggest that forces mediate distance-sensitivity of the edge dynamics and ultimately organize cell-edge behavior leading to spontaneous polarization. Actin flow rate did not exhibit the same distance-dependence as traction stress, arguing against its role in organizing edge dynamics. Finally, using a simple model of actin-myosin network, we show that force-distance relationship may be an emergent feature of such networks.

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“…We next wondered whether the polarization of actin networks could be correlated with a mechanical asymmetry during T cell migration. To answer this question, we used the dynamic Traction Force Microscopy technique (18) which allows the forces developed by the cells upon migration on gels to be measured. T cells are fast-moving cells, therefore they are expected to develop low forces on their substrate.…”

Section: Remodeling Of Actin Cytoskeleton In Chemokine-stimulated T Lmentioning

confidence: 99%

cAMP bursts control T cell directionality by actin cytoskeleton remodeling

Simao

Régnier

Taheraly

et al. 2020

Preprint

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AbstractT lymphocyte migration is an essential step to mounting an efficient immune response. The rapid and random motility of these cells which favors their sentinel role is conditioned by chemokines as well as by the physical environment. Morphological changes, underlaid by dynamic actin cytoskeleton remodeling, are observed throughout migration but especially when the cell modifies its trajectory. Using dynamic cell imaging, we investigated the signaling pathways involved in T cell directionality control. We monitored cAMP variation concomitantly with actin distribution upon T lymphocyte migration and highlighted the fact that spontaneous bursts in cAMP starting from the leading edge, are sufficient to promote stable actin redistribution triggering trajectory modification.

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Microfabricated tissues for investigating traction forces involved in cell migration and tissue morphogenesis

Cited by 26 publications

References 163 publications

A small proportion of Talin molecules transmit forces at developing muscle attachments in vivo

A small proportion of Talin molecules transmit forces at developing muscle attachments in vivo

Traction forces control cell-edge dynamics and mediate distance-sensitivity during cell polarization

cAMP bursts control T cell directionality by actin cytoskeleton remodeling

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