Chapter 22
Molecular Tension Microscopy of E-Cadherin During
Epithelial-Mesenchymal Transition
Helena Canever, Pietro Salvatore Carollo, Romain Fleurisson,
Philippe P. Girard, and Nicolas Borghi
Abstract
Molecular Tension Microscopy has been increasingly used in the last years to investigate mechanical forces
acting in cells at the molecular scale. Here, we describe a protocol to image the tension of the junctional
protein E-cadherin in cultured epithelial cells undergoing Epithelial-Mesenchymal Transition (EMT). We
report how to prepare cells and induce EMT, and how to acquire, analyze, and quantitatively interpret
FRET data.
Key words FRET Biosensor, E-cadherin, EMT, Microscopy, Mechanotransduction
1 Introduction
Molecular Tension Microscopy (MTM) is the microscopy of molecular tension sensors [1]. Molecular tension sensors are a class of
Fo ¨rster Resonance Energy Transfer (FRET) sensors that are sensitive to molecular tension. They consist in a FRET pair of fluorophores separated by an elastic linker. When a force is exerted on the
sensor, the rate of FRET, which is sensitive to the distance and
orientation between the donor and acceptor fluorophores,
decreases. Knowledge of the relationship between a measure of
FRET and the force, from in vitro calibration, allows for the determination of molecular tension [2]. When inserted in a protein of
interest, such a sensor can report its tension in live cells.
Molecular tension sensors have been in use for nearly 10 years
[3]. A variety of sensors with specific operating ranges have been
designed [4, 5], and have been used to measure tension in matrix,
cytoskeleton, adhesion, glycocalyx, kinetochore, membranecytoskeleton linker or motor proteins [2, 3, 6–9] in cultured cells,
C. elegans, Xenopus, Zebrafish, or Drosophila [3, 10–12].
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_22,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
289
Molecular Tension Microscopy of E-Cadherin During
Epithelial-Mesenchymal Transition
Helena Canever, Pietro Salvatore Carollo, Romain Fleurisson,
Philippe P. Girard, and Nicolas Borghi
Abstract
Molecular Tension Microscopy has been increasingly used in the last years to investigate mechanical forces
acting in cells at the molecular scale. Here, we describe a protocol to image the tension of the junctional
protein E-cadherin in cultured epithelial cells undergoing Epithelial-Mesenchymal Transition (EMT). We
report how to prepare cells and induce EMT, and how to acquire, analyze, and quantitatively interpret
FRET data.
Key words FRET Biosensor, E-cadherin, EMT, Microscopy, Mechanotransduction
1 Introduction
Molecular Tension Microscopy (MTM) is the microscopy of molecular tension sensors [1]. Molecular tension sensors are a class of
Fo ¨rster Resonance Energy Transfer (FRET) sensors that are sensitive to molecular tension. They consist in a FRET pair of fluorophores separated by an elastic linker. When a force is exerted on the
sensor, the rate of FRET, which is sensitive to the distance and
orientation between the donor and acceptor fluorophores,
decreases. Knowledge of the relationship between a measure of
FRET and the force, from in vitro calibration, allows for the determination of molecular tension [2]. When inserted in a protein of
interest, such a sensor can report its tension in live cells.
Molecular tension sensors have been in use for nearly 10 years
[3]. A variety of sensors with specific operating ranges have been
designed [4, 5], and have been used to measure tension in matrix,
cytoskeleton, adhesion, glycocalyx, kinetochore, membranecytoskeleton linker or motor proteins [2, 3, 6–9] in cultured cells,
C. elegans, Xenopus, Zebrafish, or Drosophila [3, 10–12].
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_22,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
289