Russian Journal of Developmental Biology

, Volume 49, Issue 6, pp 362–369 | Cite as

Development of Methods and Techniques to Visualize Mechanical Tension in Embryos Using Genetically Encoded Fluorescent Mechanosensors

  • F. M. EroshkinEmail author
  • S. V. Kremnev
  • G. V. Ermakova
  • A. G. Zaraisky


Lately, the growing body of quantitative data has provided evidence of the importance of mechanical forces in embryogenesis. The study of spatial and temporal distribution of mechanical tension in the course of embryogenesis is one of the most important problems of modern developmental biology. Development of genetically encoded fluorescent mechanosensors allowed their application in an intravital study of mechanical tension in developing embryos via noninvasive techniques. The possibility of applying fluorescent mechanosensors based on vinculin and C-cadherin to visualize mechanical tension in tissues of Gallus and Xenopus embryos was studied. The methods to express and detect these proteins, as well as process the resulting images, were elaborated. The best results were obtained using Xenopus embryos and the vinculin-based mechanosensor.


Xenopus Gallus embryogenesis mechaniсal tension mechanosensors 



The work was supported by the Russian Foundation for Basic Research (project no. 15-04-06310). Works on cloning of plasmid constructs were supported by the Russian Science Foundation (project no. 14-50-00131).


  1. 1.
    Beloussov, L.V., Mechanically based generative laws of morphogenesis, Phys. Biol., 2008, vol. 5, p. 015009.CrossRefGoogle Scholar
  2. 2.
    Beloussov, L.V., Dorfman, J.G., and Cherdantzev, V.G., Mechanical stresses and morphological patterns in amphibian embryos, J. Embryol. Exp. Morphol., 1975, vol. 34, pp. 559–574.Google Scholar
  3. 3.
    Beloussov, L.V., Lakirev, A.V., Naumidi, I.I., et al., Effects of relaxation of mechanical tensions upon the early morphogenesis of Xenopus laevis embryos, Int. J. Dev. Biol., 1990, vol. 34, pp. 409–419.Google Scholar
  4. 4.
    Borghi, N., Sorokina, M., Shcherbakova, O.G., et al., E‑cadherin is under constitutive actomyosin-generated tension that is increased at cell-cell contacts upon externally applied stretch, Proc. Natl. Acad. Sci. U. S. A., 2012, vol. 109, pp. 12568–12573.CrossRefGoogle Scholar
  5. 5.
    Conway, D.E., Breckenridge, M.T., Hinde, E., et al., Fluid shear stress on endothelial cells modulates mechanical tension across VE-cadherin and PECAM-1, Curr. Biol., 2013, vol. 23, pp. 1024–1030.CrossRefGoogle Scholar
  6. 6.
    Dong, H.M., Liu, G., Hou, Y.F., et al., Dominant-negative E-cadherin inhibits the invasiveness of inflammatory breast cancer cells in vitro, J. Cancer Res. Clin. Oncol., 2007, vol. 133, pp. 83–92.CrossRefGoogle Scholar
  7. 7.
    Eroshkin, F.M. and Zaraisky, A.G., Mechano-sensitive regulation of gene expression during the embryonic development, Genesis, 2017, vol. 55.Google Scholar
  8. 8.
    Eyckmans, J., Boudou, T., Yu, X., et al., A hitchhiker’s guide to mechanobiology, Dev. Cell, 2011, vol. 21, pp. 35–47.CrossRefGoogle Scholar
  9. 9.
    Grashoff, C., Hoffman, B.D., Brenner, M.D., et al., Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics, Nature, 2010, vol. 466, pp. 263–266.CrossRefGoogle Scholar
  10. 10.
    Guo, J., Sachs, F., and Meng, F., Fluorescence-based force/tension sensors: a novel tool to visualize mechanical forces in structural proteins in live cells, Antioxid. Redox Signal., 2014, vol. 20, pp. 986–999.CrossRefGoogle Scholar
  11. 11.
    Ishibashi, S., Love, N.R., and Amaya, E., A simple method of transgenesis using I-SceI meganuclease in Xenopus, Methods Mol. Biol., 2012, vol. 917, pp. 205–218.CrossRefGoogle Scholar
  12. 12.
    Kroll, K.L. and Amaya, E., Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation, Development, 1996, vol. 122, pp. 3173–3183.Google Scholar
  13. 13.
    Lee, C.H. and Gumbiner, B.M., Disruption of gastrulation movements in Xenopus by a dominant-negative mutant for C-cadherin, Dev. Biol., 1995, vol. 171, pp. 363–373.CrossRefGoogle Scholar
  14. 14.
    Martynova, N.Y., Eroshkin, F.M., Ermolina, L.V., et al., The LIM-domain protein Zyxin binds the homeodomain factor Xanf1/Hesx1 and modulates its activity in the anterior neural plate of Xenopus laevis embryo, Dev. Dyn., 2008, vol. 237, pp. 736–749.CrossRefGoogle Scholar
  15. 15.
    Nandadasa, S., Tao, Q., Menon, N.R., et al., N- and E‑cadherins in Xenopus are specifically required in the neural and non-neural ectoderm, respectively, for F-actin assembly and morphogenetic movements, Development, 2009, vol. 136, pp. 1327–1338.CrossRefGoogle Scholar
  16. 16.
    Ogino, H., McConnell, W.B., and Grainger, R.M., High-throughput transgenesis in Xenopus using I-SceI meganuclease, Nat Protoc, 2006a, vol. 1, pp. 1703–1710.CrossRefGoogle Scholar
  17. 17.
    Ogino, H., McConnell, W.B., and Grainger, R.M., Highly efficient transgenesis in Xenopus tropicalis using I-SceI meganuclease, Mech. Dev., 2006b, vol. 123, pp. 103–113.CrossRefGoogle Scholar
  18. 18.
    Rahimzadeh, J., Meng, F., Sachs, F., et al., Real-time observation of flow-induced cytoskeletal stress in living cells, Am. J. Physiol. Cell Physiol., 2011, vol. 301, pp. 646–652.CrossRefGoogle Scholar
  19. 19.
    Stooke-Vaughan, G.A., Davidson, L.A., and Woolner, S., Xenopus as a model for studies in mechanical stress and cell division, Genesis, 2017, vol. 55.Google Scholar
  20. 20.
    Vizirianakis, I.S., Chen, Y.Q., Kantak, S.S., et al., Dominant-negative E-cadherin alters adhesion and reverses contact inhibition of growth in breast carcinoma cells, Int. J. Oncol., 2002, vol. 21, pp. 135–144.Google Scholar
  21. 21.
    Yamashita, S., Tsuboi, T., Ishinabe, N., Kitaguchi, T., and Michiue, T., Wide and high resolution tension measurement using fret in embryo, Sci. Rep., 2016, vol. 6, p. 28535.CrossRefGoogle Scholar
  22. 22.
    Yang, C., Zhang, X., Guo, Y., Meng, F., Sachs, F., and Guo, J., Mechanical dynamics in live cells and fluorescence-based force/tension sensors, Biochim. Biophys. Acta, 2015, vol. 1853, pp. 1889–1904.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Inc. 2018

Authors and Affiliations

  • F. M. Eroshkin
    • 1
    Email author
  • S. V. Kremnev
    • 2
    • 3
  • G. V. Ermakova
    • 1
  • A. G. Zaraisky
    • 1
  1. 1.Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of SciencesMoscowRussia
  2. 2.Department of Embryology, Faculty of Biology, Moscow State UniversityMoscowRussia
  3. 3.Koltsov Institute of Developmental Biology, Russian Academy of SciencesMoscowRussia

Personalised recommendations