Lighting Up the Force: Investigating Mechanisms of Mechanotransduction Using Fluorescent Tension Probes

Jurchenko, Carol; Salaita, Khalid

doi:10.1128/mcb.00195-15

Cited by 78 publications

(73 citation statements)

References 145 publications

(161 reference statements)

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“…Molecular tension fluorescence microscopy (MTFM, previously reviewed by Jurchenko and Salaita [131]) is a method to optically image receptor mechanics at the living-nonliving interface. Previously, this method has been applied to map T cell receptor, epidermal growth factor receptor, and integrin forces with high spatiotemporal resolution [11, 15, 131–136].…”

Section: Methods To Measure Receptor Forces At Fluid Interfacesmentioning

confidence: 99%

“…Previously, this method has been applied to map T cell receptor, epidermal growth factor receptor, and integrin forces with high spatiotemporal resolution [11, 15, 131–136]. In MTFM, an immobilized probe molecule comprised of a flexible linker and flanked by a fluorephore-quencher pair presents a ligand to a receptor of interest.…”

Section: Methods To Measure Receptor Forces At Fluid Interfacesmentioning

confidence: 99%

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Supported lipid bilayer platforms to probe cell mechanobiology

Glazier¹,

Salaita²

2017

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Mammalian and bacterial cells sense and exert mechanical forces through the process of mechanotransduction, which interconverts biochemical and physical signals. This is especially important in contact-dependent signaling, where ligand-receptor binding occurs at cell-cell or cell-ECM junctions. By virtue of occurring within these specialized junctions, receptors engaged in contact-dependent signaling undergo oligomerization and coupling with the cytoskeleton as part of their signaling mechanisms. While our ability to measure and map biochemical signaling within cell junctions has advanced over the past decades, physical cues remain difficult to map in space and time. Recently, supported lipid bilayer (SLB) technologies have emerged as a flexible platform to measure and manipulate membrane receptor mechanotransduction, allowing one to mimic cell-cell junctions. Changing the lipid composition and underlying substrate tunes bilayer fluidity, and lipid and ligand micro- and nano-patterning spatially control positioning and clustering of receptors. Patterning metal gridlines within SLBs introduces corrals that confine lipid mobility and introduce mechanical resistance. Here we review fundamental SLB mechanics and how SLBs can be engineered as tunable cell substrates for mechanotransduction studies. Finally, we highlight the impact of this work in understanding the biophysical mechanisms of cell adhesion.

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Section: Methods To Measure Receptor Forces At Fluid Interfacesmentioning

confidence: 99%

Section: Methods To Measure Receptor Forces At Fluid Interfacesmentioning

confidence: 99%

Supported lipid bilayer platforms to probe cell mechanobiology

Glazier¹,

Salaita²

2017

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…[219–221]. The technique consists in inserting a molecular spacer with known extensional rigidity (spring constant) in between the donor and acceptor of a FRET system.…”

Section: Methodologies To Probe the Mechanics Of Living Embryonic mentioning

confidence: 99%

A toolbox to explore the mechanics of living embryonic tissues

Campàs

2016

Seminars in Cell & Developmental Biology

128

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The sculpting of embryonic tissues and organs into their functional morphologies involves the spatial and temporal regulation of mechanics at cell and tissue scales. Decades of in vitro work, complemented by some in vivo studies, have shown the relevance of mechanical cues in the control of cell behaviors that are central to developmental processes, but the lack of methodologies enabling precise, quantitative measurements of mechanical cues in vivo have hindered our understanding of the role of mechanics in embryonic development. Several methodologies are starting to enable quantitative studies of mechanics in vivo and in situ, opening new avenues to explore how mechanics contributes to shaping embryonic tissues and how it affects cell behavior within developing embryos. Here we review the present methodologies to study the role of mechanics in living embryonic tissues, considering their strengths and drawbacks as well as the conditions in which they are most suitable.

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“…[3] The initial tension probes were comprised of an extendable polyethylene glycol (PEG) spring, flanked by a fluorophore and a spectroscopically-matched quencher. [4] pN forces extend the mean end-to-end distance of the polymer, which reduces energy transfer through an R −6 distance-dependent relationship.…”

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confidence: 99%

Mechanically Induced Catalytic Amplification Reaction for Readout of Receptor‐Mediated Cellular Forces

et al. 2016

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Mechanics play a fundamental role in cell biology, but detecting piconewton forces is challenging due to the lack of accessible and high throughput assays. Herein we report the mechanically-induced catalytic amplification reaction (MCR) for readout of receptor-mediated forces in cells. Mechanically labile DNA duplexes presenting ligands are surface immobilized such that specific receptor forces denature the duplex and thus expose a blocked primer. Amplification of primers is achieved using an isothermal polymerization reaction and quantified by fluorescence readout. As a proof-of-concept, the assay was used to test the activity of a mechano-modulatory drug and integrin adhesion receptor antibodies. To the best of our knowledge, this is the first example of a catalytic reaction triggered in response to molecular piconewton forces. The MCR may transform the field of mechanobiology by providing a new facile tool to detect receptor-specific mechanics with the convenience of the PCR.

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Lighting Up the Force: Investigating Mechanisms of Mechanotransduction Using Fluorescent Tension Probes

Cited by 78 publications

References 145 publications

Supported lipid bilayer platforms to probe cell mechanobiology

Supported lipid bilayer platforms to probe cell mechanobiology

A toolbox to explore the mechanics of living embryonic tissues

Mechanically Induced Catalytic Amplification Reaction for Readout of Receptor‐Mediated Cellular Forces

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