More Like this

1. Vinculin tension probe in neurons

   https://doi.org/10.1117/12.2578501  

   Ayad, Marina A.; Mahon, Timothy; Sumetsky, Daniel; Cararo-Lopes, Marina M.; Firestein, Bonnie L.; Boustany, Nada N. (March 2021, Optical Techniques in Neurosurgery, Neurophotonics, and Optogenetics)

   Yang, Victor X.; Kainerstorfer, Jana M.; Luo, Qingming; Ding, Jun; Fu, Ling; Mohanty, Samarendra K.; Roe, Anna W.; Shoham, Shy (Ed.)

   Vinculin is a known key regulator of focal adhesions; it undergoes tension in the locations of attachment to the extracellular matrix. In this study, we explore the use of a vinculin tension FRET probe to investigate vinculin tension within neurons. A critical component of neuronal growth is migration, which is dependent on the mechanical cues between the cells and the extracellular matrix. An understanding of tension variation within the neuron may help us understand mechanisms of neurogenesis. To study these forces, we use a previously developed molecular tension sensor, which consists of an elastic linker, TSMod, a 40-amino-acid-long peptide inserted between teal fluorescent protein (mTFP1) and mVenus. The vinculin tension sensor, VinTS, consists of TSMod embedded between the Vinculin head and tail. When under tension, VinTS will exhibit a lower fluorescence resonance energy transfer (FRET) efficiency between mTFP1 and mVenus. Cortical neurons were isolated from embryonic rat brains and cultured on glass coverslips coated with poly-D-lysine and laminin. The neurons were transfected with TSMod (the unloaded tension sensor) or VinTS. Neurons expressing TSMod are used as the experiment’s control group since TSMod on its own is not affected by vinculin tension. The mean FRET efficiency of 171 TSMod and 127 VinTS expressing neurons was 27.08 ± 4.98%, and 22.86 ± 3.98%, respectively. The FRET efficiency of VinTS was significantly lower than that of TSMod (p = 6.6e15 by Welch’s t-test). These results support the feasibility of using the VinTS probe in neurons and provide a first assessment of VinTS FRET efficiency in neurons. The lower FRET efficiency of VinTS compared with TSMod suggests that VinTS may be under tension in neurons. However, additional studies are required to further characterize these results. 

   more » « less
2. α–Catenin-dependent vinculin recruitment to adherens junctions is antagonistic to focal adhesions

   https://doi.org/10.1091/mbc.E22-02-0071  

   Bejar-Padilla, Vidal; Cabe, Jolene I.; Lopez, Santiago; Narayanan, Vani; Mezher, Mazen; Maruthamuthu, Venkat; Conway, Daniel E. (September 2022, Molecular Biology of the Cell)

   Yap, Alpha (Ed.)

   Vinculin is a protein found in both focal adhesions (FAs) and adherens junctions (AJs) which regulates actin connectivity to these structures. Many studies have demonstrated that mechanical perturbations of cells result in enhanced recruitment of vinculin to FAs and/or AJs. Likewise, many other studies have shown “cross-talk” between FAs and AJs. Vinculin itself has been suggested to be a probable regulator of this adhesion cross-talk. In this study we used MDCK as a model system of epithelia, developing cell lines in which vinculin recruitment was reduced or enhanced at AJs. Careful analysis of these cells revealed that perturbing vinculin recruitment to AJs resulted in a reduction of detectable FAs. Interestingly the cross-talk between these two structures was not due to a limited pool of vinculin, as increasing expression of vinculin did not rescue FA formation. Instead, we demonstrate that vinculin translocation between AJs and FAs is necessary for actin cytoskeleton rearrangements that occur during cell migration, which is necessary for large, well-formed FAs. Last, we show using a wound assay that collective cell migration is similarly hindered when vinculin recruitment is reduced or enhanced at AJs, highlighting that vinculin translocation between each compartment is necessary for efficient collective migration. 

   more » « less
3. Vinculin transmits high-level integrin tensions that are dispensable for focal adhesion formation

   https://doi.org/10.1016/j.bpj.2022.11.013  

   Austin, Jacob; Tu, Ying; Pal, Kaushik; Wang, Xuefeng (January 2023, Biophysical Journal)
4. Cyclic stretch regulates epithelial cell migration in a frequency dependent manner via intercellular vinculin recruitment

   https://doi.org/10.1038/s44341-024-00003-1  

   Dow, Liam_P; Surace, Stacey; Morozov, Katrene; Kennedy, Reagan; Pruitt, Beth_L (November 2024, npj Biological Physics and Mechanics)

   Abstract The epithelial microenvironment is incredibly dynamic, subjected to mechanical cues including cyclic stretch. While cyclic cell stretching platforms have revealed epithelial cell reorientation and gap formation, few studies have investigated the long-term effects of cyclic stretch on cell migration. We measured the migratory response of the epithelium to a range of physiologically relevant frequencies and stretch. Our results indicate that lower stretch frequencies (i.e., 0.1 Hz) suppress epithelial migration, accompanied by cell reorientation and high cell shape solidity. We found that this response is also accompanied by increased recruitment of vinculin to cell-cell contacts, and this recruitment is necessary to suppress cell movements. These results confirm the mechanosensitive nature of vinculin within the adherens junction, but independently reveal a novel mechanism of low frequency stress response in supporting epithelial integrity by suppressing cell migration. 

   more » « less
5. Nanoscale dynamics of the cadherin–catenin complex bound to vinculin revealed by neutron spin echo spectroscopy

   https://doi.org/10.1073/pnas.2408459121  

   Callaway, David_J E; Nicholl, Iain D; Shi, Bright; Reyes, Gilbert; Farago, Bela; Bu, Zimei (September 2024, Proceedings of the National Academy of Sciences)

   Weitz, David (Ed.)

   We report a neutron spin echo (NSE) study of the nanoscale dynamics of the cell–cell adhesion cadherin–catenin complex bound to vinculin. Our measurements and theoretical physics analyses of the NSE data reveal that the dynamics of full-length α-catenin, β-catenin, and vinculin residing in the cadherin–catenin–vinculin complex become activated, involving nanoscale motions in this complex. The cadherin–catenin complex is the central component of the cell–cell adherens junction (AJ) and is fundamental to embryogenesis, tissue wound healing, neuronal plasticity, cancer metastasis, and cardiovascular health and disease. A highly dynamic cadherin–catenin–vinculin complex provides the molecular dynamics basis for the flexibility and elasticity that are necessary for the AJs to function as force transducers. Our theoretical physics analysis provides a way to elucidate these driving nanoscale motions within the complex without requiring large-scale numerical simulations, providing insights not accessible by other techniques. We propose a three-way “motorman” entropic spring model for the dynamic cadherin–catenin–vinculin complex, which allows the complex to function as a flexible and elastic force transducer. 

   more » « less