Laser Tweezers in the Study of Mechanobiology in Live Cells

Botvinick, Elliot L.; Wang, Yingxiao

doi:10.1016/s0091-679x(06)82018-2

Cited by 8 publications

(6 citation statements)

References 25 publications

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“…Single-beam gradient optical laser tweezers with controlled 300 pN of mechanical force were applied to pull the adhered beads. A similar optical trapping system has been described in our previous report ( Botvinick and Wang, 2007 ).…”

Section: Methodsmentioning

confidence: 99%

“…Various mechanical stimulations can affect cytosolic Ca 2+ signals as well as Ca 2+ dynamics in organelles or subcellular compartments, such as mitochondria and focal adhesion sites ( Balasubramanian et al, 2007 ; Belmonte and Morad, 2008 ; Hayakawa et al, 2008 ; Horner and Wolfner, 2008 ). To apply mechanical force precisely, we utilize optical laser tweezers to generate force in a bead coupled to the cell surface through the ligation of adhesion receptors and transmit it into the cell at subcellular locations to trigger signal transduction ( Berns, 2007 ; Botvinick and Wang, 2007 ).…”

Section: Introductionmentioning

confidence: 99%

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Distinct mechanisms regulating mechanical force-induced Ca2+ signals at the plasma membrane and the ER in human MSCs

Kim

Joo

Seong

et al. 2015

eLife

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It is unclear that how subcellular organelles respond to external mechanical stimuli. Here, we investigated the molecular mechanisms by which mechanical force regulates Ca2+ signaling at endoplasmic reticulum (ER) in human mesenchymal stem cells. Without extracellular Ca2+, ER Ca2+ release is the source of intracellular Ca2+ oscillations induced by laser-tweezer-traction at the plasma membrane, providing a model to study how mechanical stimuli can be transmitted deep inside the cell body. This ER Ca2+ release upon mechanical stimulation is mediated not only by the mechanical support of cytoskeleton and actomyosin contractility, but also by mechanosensitive Ca2+ permeable channels on the plasma membrane, specifically TRPM7. However, Ca2+ influx at the plasma membrane via mechanosensitive Ca2+ permeable channels is only mediated by the passive cytoskeletal structure but not active actomyosin contractility. Thus, active actomyosin contractility is essential for the response of ER to the external mechanical stimuli, distinct from the mechanical regulation at the plasma membrane.DOI: http://dx.doi.org/10.7554/eLife.04876.001

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Distinct mechanisms regulating mechanical force-induced Ca2+ signals at the plasma membrane and the ER in human MSCs

Kim

Joo

Seong

et al. 2015

eLife

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“…This improved biosensor displayed a high sensitivity and specificity toward Src kinase, and allowed the successful visualization of a wave propagation of Src activation upon mechanical stimulation induced by a laser-tweezer-traction of a bead seeded on top of human umbilical vein endothelial cell (HUVECs). 5,52 Recently, ECFP and YPet were shown to serve as an improved FRET pair to allow a significantly enhanced dynamic range for various FRET biosensors, including the Src FRET biosensor. 41 This third version of Src biosensor with enhanced sensitivity can allow the detection of subtle, but physiologically crucial signals.…”

Section: Live Cell Imaging Of the Src/fak Signaling By Fretmentioning

confidence: 99%

Live Cell Imaging of Src/FAK Signaling by FRET

Wang

2011

Cel. Mol. Bioeng.

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The Src/FAK complex is involved in many signaling pathways and plays crucial roles in cell adhesion/migration. It becomes clear that the subcellular localization of Src and FAK is crucial for their activities and functions. In this article, we first overview the molecular mechanisms and functions of Src and FAK involved in cell adhesion/migration. We then introduce the development of genetically encoded biosensors based on fluorescence resonance energy transfer (FRET) to visualize the activities of Src and FAK in live cells with high spatiotemporal resolutions. Different kinds of signal peptides targeting subcellular compartments are also discussed. FRET-based biosensors fused with these targeting signals peptides are further introduced to provide an overview on how these targeting signals can facilitate the localization of biosensors to continuously monitor the local activity of Src and FAK at subcellular compartments. In summary, genetically-encoded FRET biosensors integrated with subcellular compartment-targeting signals can provide powerful tools for the visualization of subcellular Src and FAK activities in live cells and advance our in-depth understanding of Src/FAK functions at different subcellular compartments.

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“…The development of genetically encoded FRET-based reporters allows the investigation of different mechanically induced signals with high sensitivity within living cells but it is a challenging process due to the need of calibration and control experiments. A first step in this direction was taken by Wang et al who developed a FRET-based reporter for the activation of the Src protein, a regulator of the integrin-cytoskeleton interaction (Botvinick and Wang 2007;Wang et al 2005). Application of a pulling force via laser tweezers on human umbilical vein endothelial cells (HUVECs) expressing the membrane-targeted Src reporter led to a directional FRET response, with the majority of activations transmitted towards distal areas of the cell opposite to the force direction (Botvinick and Wang 2007;Wang et al 2005).…”

Section: Measuring Mechanotransduction Signals In Living Cellsmentioning

confidence: 99%

Probing force in living cells with optical tweezers: from single-molecule mechanics to cell mechanotransduction

Arbore¹,

Perego²,

Σεργίδης³

et al. 2019

Biophys Rev

109

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The invention of optical tweezers more than three decades ago has opened new avenues in the study of the mechanical properties of biological molecules and cells. Quantitative force measurements still represent a challenging task in living cells due to the complexity of the cellular environment. Here, we review different methodologies to quantitatively measure the mechanical properties of living cells, the strength of adhesion/receptor bonds, and the active force produced during intracellular transport, cell adhesion, and migration. We discuss experimental strategies to attain proper calibration of optical tweezers and molecular resolution in living cells. Finally, we show recent studies on the transduction of mechanical stimuli into biomolecular and genetic signals that play a critical role in cell health and disease.

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Laser Tweezers in the Study of Mechanobiology in Live Cells

Cited by 8 publications

References 25 publications

Distinct mechanisms regulating mechanical force-induced Ca2+ signals at the plasma membrane and the ER in human MSCs

Distinct mechanisms regulating mechanical force-induced Ca2+ signals at the plasma membrane and the ER in human MSCs

Live Cell Imaging of Src/FAK Signaling by FRET

Probing force in living cells with optical tweezers: from single-molecule mechanics to cell mechanotransduction

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