Decision letter: Analysis of cellular behavior and cytoskeletal dynamics reveal a constriction mechanism driving optic cup morphogenesis

Analysis of cellular behavior and cytoskeletal dynamics reveal a constriction mechanism driving optic cup morphogenesis

eLife ◽

10.7554/elife.15797 ◽

2016 ◽

Vol 5 ◽

Cited By ~ 33

Author(s):

María Nicolás-Pérez ◽

Franz Kuchling ◽

Joaquín Letelier ◽

Rocío Polvillo ◽

Jochen Wittbrodt ◽

...

Keyword(s):

Laser Cutting ◽

Mechanical Tension ◽

Cellular Behavior ◽

Optic Cup ◽

Tensile Forces ◽

Cytoskeletal Dynamics ◽

Cellular Machinery ◽

Vertebrate Eye ◽

Fast Oscillations ◽

Developmental Window

Contractile actomyosin networks have been shown to power tissue morphogenesis. Although the basic cellular machinery generating mechanical tension appears largely conserved, tensions propagate in unique ways within each tissue. Here we use the vertebrate eye as a paradigm to investigate how tensions are generated and transmitted during the folding of a neuroepithelial layer. We record membrane pulsatile behavior and actomyosin dynamics during zebrafish optic cup morphogenesis by live imaging. We show that retinal neuroblasts undergo fast oscillations and that myosin condensation correlates with episodic contractions that progressively reduce basal feet area. Interference with lamc1 function impairs basal contractility and optic cup folding. Mapping of tensile forces by laser cutting uncover a developmental window in which local ablations trigger the displacement of the entire tissue. Our work shows that optic cup morphogenesis is driven by a constriction mechanism and indicates that supra-cellular transmission of mechanical tension depends on ECM attachment.

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Author response: Analysis of cellular behavior and cytoskeletal dynamics reveal a constriction mechanism driving optic cup morphogenesis

10.7554/elife.15797.039 ◽

2016 ◽

Author(s):

María Nicolás-Pérez ◽

Franz Kuchling ◽

Joaquín Letelier ◽

Rocío Polvillo ◽

Jochen Wittbrodt ◽

...

Keyword(s):

Author Response ◽

Response Analysis ◽

Cellular Behavior ◽

Optic Cup ◽

Cytoskeletal Dynamics

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A Toolkit for Manipulating Cellular Behavior: Peptide with Tryptophan-Selective Ru-TAP Complex Regioselectively Photolabeling Specific Protein in Live Cell

10.21203/rs.3.rs-1073094/v1 ◽

2021 ◽

Author(s):

Tzu-Ho Chen ◽

Kevin Garnir ◽

Chong-Yen Chen ◽

Cheng-Bang Jian ◽

Hua-De Gao ◽

...

Keyword(s):

Blue Light ◽

Photoinduced Electron Transfer ◽

Protein Modification ◽

High Selectivity ◽

Specific Protein ◽

Protein Labeling ◽

Cellular Behavior ◽

Native Proteins ◽

Endogenous Protein ◽

Cytoskeletal Dynamics

Abstract Using a chemical approach to crosslink functionally versatile bioeffectors (such as peptides) to native proteins of interest (POI) directly inside a living cell is a useful toolbox for chemical biologists. However, this goal has not been reached due to unsatisfactory chemoselectivity, regioselectivity, and protein-selectivity in in-cellulo protein labeling. Herein we report a highly selective photoaffinity labeling (PAL) method using a tryptophan-specific Ru-TAP complex as photocrosslinker (Trp-tag). Aside from the high selectivity, the PAL is blue light driven by a photoinduced electron transfer (PeT) and allows the bioeffector to bear an additional UV-responsive unit. The two different photosensitivities are demonstrated by blue light photocrosslinking a UV-sensitive peptide to POI. The remote-control functionality of the peptide allows POI inhibition after blue light irradiation, and reactivation upon UV photolysis. Cytoskeletal dynamics regulation is demonstrated via the unprecedented in-cellulo POI photomanipulation, which opens a new avenue to endogenous protein modification for novel functions.

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Use of Flexible Materials with a Novel Pressure Driven Equibiaxial Cell Stretching Device for Mechanical Stimulation of Single Mammalian Cells

MRS Proceedings ◽

10.1557/proc-773-n5.6 ◽

2003 ◽

Vol 773 ◽

Author(s):

James D. Kubicek ◽

Stephanie Brelsford ◽

Philip R. LeDuc

Keyword(s):

Cell Fate ◽

Mechanical Stimulation ◽

Mammalian Cells ◽

Cellular Response ◽

Single Cells ◽

Complex Nature ◽

Cellular Behavior ◽

Cell Stretching ◽

Stimulation Of ◽

Pressure Driven

AbstractMechanical stimulation of single cells has been shown to affect cellular behavior from the molecular scale to ultimate cell fate including apoptosis and proliferation. In this, the ability to control the spatiotemporal application of force on cells through their extracellular matrix connections is critical to understand the cellular response of mechanotransduction. Here, we develop and utilize a novel pressure-driven equibiaxial cell stretching device (PECS) combined with an elastomeric material to control specifically the mechanical stimulation on single cells. Cells were cultured on silicone membranes coated with molecular matrices and then a uniform pressure was introduced to the opposite surface of the membrane to stretch single cells equibiaxially. This allowed us to apply mechanical deformation to investigate the complex nature of cell shape and structure. These results will enhance our knowledge of cellular and molecular function as well as provide insights into fields including biomechanics, tissue engineering, and drug discovery.

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A Photoactivatable α5β1-Specific Integrin Ligand

10.26434/chemrxiv.6097688 ◽

2018 ◽

Author(s):

Roshna Vakkeel ◽

Aleeza Farrukh ◽

Aranzazu del Campo

Keyword(s):

Cellular Response ◽

Light Exposure ◽

Matrix Composition ◽

Cellular Behavior ◽

Dynamic Variations ◽

In Situ Activation ◽

Controlled Exposure ◽

Cellular Behaviour ◽

Integrin Ligand

In order to study how dynamic changes of α5β1 integrin engagement affect cellular behaviour, photoactivatable derivatives of α5β1 specific ligands are presented in this article. The presence of the photoremovable protecting group (PRPG) introduced at a relevant position for integrin recognition, temporally inhibits ligand bioactivity. Light exposure at cell-compatible dose efficiently cleaves the PRPG and restores functionality. Selective cell response (attachment, spreading, migration) to the activated ligand on the surface is achieved upon controlled exposure. Spatial and temporal control of the cellular response is demonstrated, including the possibility to in situ activation. Photoactivatable integrin-selective ligands in model microenvironments will allow the study of cellular behavior in response to changes in the activation of individual integrins as consequence of dynamic variations of matrix composition.

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