Molecular tension indicators reveal unexpectedly complex regulation of tension in live mouse organs

1. Grashoff, C. et al. Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics. Nature 466, 263–266 (2010).

   Google Scholar 

2. LaCroix, A. S., Lynch, A. D., Berginski, M. E. & Hoffman, B. D. Tunable molecular tension sensors reveal extension-based control of vinculin loading. eLife 7, e33927 (2018).

   Google Scholar 

3. Déjardin, T. et al. Nesprins are mechanotransducers that discriminate epithelial–mesenchymal transition programs. J. Cell Biol. 219, e201908036 (2020).

   Google Scholar 

4. Kim, T.-J. et al. Dynamic visualization of α-catenin reveals rapid, reversible conformation switching between tension states. Curr. Biol. 25, 218–224 (2015).

   Google Scholar 

5. Kumar, A. et al. Talin tension sensor reveals novel features of focal adhesion force transmission and mechanosensitivity. J. Cell Biol. 213, 371–383 (2016).

   Google Scholar 

6. Lagendijk, A. K. et al. Live imaging molecular changes in junctional tension upon VE-cadherin in zebrafish. Nat. Commun. 8, 1402 (2017).

   Google Scholar 

7. Morimatsu, M., Mekhdjian, A. H., Adhikari, A. S. & Dunn, A. R. Molecular tension sensors report forces generated by single integrin molecules in living cells. Nano Lett. 13, 3985–3989 (2013).

   Google Scholar 

8. Yamashita, S., Tsuboi, T., Ishinabe, N., Kitaguchi, T. & Michiue, T. Wide and high resolution tension measurement using FRET in embryo. Sci. Rep. 6, 28535 (2016).

   Google Scholar 

9. Lemke, S. B., Weidemann, T., Cost, A.-L., Grashoff, C. & Schnorrer, F. A small proportion of Talin molecules transmit forces at developing muscle attachments in vivo. PLOS Biol. 17, e3000057 (2019).

   Google Scholar 

10. Wang, J. et al. In situ FRET measurement of cellular tension using conventional confocal laser microscopy in newly established reporter mice expressing actinin tension sensor. Sci. Rep. 13, 22729 (2023).

    Google Scholar 

11. Sanfeliu-Cerdán, N., Lin, L.-C., Dunn, A. R., Goodman, M. B. & Krieg, M. Visualizing neurons under tension in vivo with optogenetic molecular force sensors. Methods Mol. Biol. 2600, 239–266 (2023).

    Google Scholar 

12. Eder, D., Basler, K. & Aegerter, C. M. Challenging FRET-based E-Cadherin force measurements in Drosophila. Sci. Rep. 7, 13692 (2017).

    Google Scholar 

13. Ham, T. R., Collins, K. L. & Hoffman, B. D. Molecular tension sensors: moving beyond force. Curr. Opin. Biomed. Eng. 12, 83–94 (2019).

    Google Scholar 

14. Baird, G. S., Zacharias, D. A. & Tsien, R. Y. Circular permutation and receptor insertion within green fluorescent proteins. Proc. Natl. Acad. Sci. 96, 11241–11246 (1999).

    Google Scholar 

15. Nakai, J., Ohkura, M. & Imoto, K. A high signal-to-noise Ca2+ probe composed of a single green fluorescent protein. Nat. Biotechnol. 19, 137–141 (2001).

    Google Scholar 

16. Ohkura, M. et al. Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals. PLoS ONE 7, e51286 (2012).

    Google Scholar 

17. Yonemura, S., Wada, Y., Watanabe, T., Nagafuchi, A. & Shibata, M. α-Catenin as a tension transducer that induces adherens junction development. Nat. Cell Biol. 12, 533–542 (2010).

    Google Scholar 

18. Maki, K. et al. Mechano-adaptive sensory mechanism of α-catenin under tension. Sci. Rep. 6, 24878 (2016).

    Google Scholar 

19. Meng, F. & Sachs, F. Visualizing dynamic cytoplasmic forces with a compliance-matched FRET sensor. J. Cell Sci. 124, 261–269 (2011).

    Google Scholar 

20. Várkuti, B. H. et al. A highly soluble, non-phototoxic, non-fluorescent blebbistatin derivative. Sci. Rep. 6, 26141 (2016).

    Google Scholar 

21. Aberle, H. et al. Assembly of the cadherin-catenin complex in vitro with recombinant proteins. J. Cell Sci. 107, 3655–3663 (1994).

    Google Scholar 

22. Pokutta, S. & Weis, W. I. Structure of the dimerization and β-catenin- binding region of α-catenin. Mol. Cell 5, 533–543 (2000).

    Google Scholar 

23. Tu, Y., Pal, K., Austin, J. & Wang, X. Filopodial adhesive force in discrete nodes revealed by integrin molecular tension imaging. Curr. Biol. 32, 4386–4396.e3 (2022).

    Google Scholar 

24. Hippenmeyer, S. et al. Genetic mosaic dissection of Lis1 and Ndel1 in neuronal migration. Neuron 68, 695–709 (2010).

    Google Scholar 

25. Jiao, K. et al. An essential role of Bmp4 in the atrioventricular septation of the mouse heart. Genes Dev. 17, 2362–2367 (2003).

    Google Scholar 

26. Niwa, H. et al. An efficient gene-trap method using poly A trap vectors and characterization of gene-trap events1. J. Biochem. 113, 343–349 (1993).

    Google Scholar 

27. Horiguchi, M. et al. Fibulin-4 conducts proper elastogenesis via interaction with cross-linking enzyme lysyl oxidase. Proc. Natl. Acad. Sci. 106, 19029–19034 (2009).

    Google Scholar 

28. Dietz, H. & Rief, M. Exploring the energy landscape of GFP by single-molecule mechanical experiments. Proc. Natl. Acad. Sci. 101, 16192–16197 (2004).

    Google Scholar 

29. Mickler, M. et al. Revealing the bifurcation in the unfolding pathways of GFP by using single-molecule experiments and simulations. Proc. Natl. Acad. Sci. 104, 20268–20273 (2007).

    Google Scholar 

30. Fischer, L. S., Rangarajan, S., Sadhanasatish, T. & Grashoff, C. Molecular force measurement with tension sensors. Annu. Rev. Biophys. 50, 595–616 (2021).

    Google Scholar 

31. Hsu, P. D. et al. DNA targeting specificity of RNA-guided Cas9 nucleases. Nat. Biotechnol. 31, 827–832 (2013).

    Google Scholar 

32. Yin, J. et al. Genetically encoded short peptide tag for versatile protein labeling by Sfp phosphopantetheinyl transferase. Proc. Natl. Acad. Sci. 102, 15815–15820 (2005).

    Google Scholar 

33. Matsuda, A., Schermelleh, L., Hirano, Y., Haraguchi, T. & Hiraoka, Y. Accurate and fi ducial-marker-free correction forthree-dimensional chromatic shift in biological fl uorescence microscopy. Sci. Rep. 8, 7583 (2018).

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Authors and Affiliations

1. Department of Pharmacology, Kansai Medical University, Hirakata, Osaka, Japan

   Keita Fujiwara, Tomoya O. Akama, Tomoyuki Nakamura & Maretoshi Hirai

2. Department of Medicine II, Kansai Medical University, Hirakata, Osaka, Japan

   Keita Fujiwara & Ichiro Shiojima

3. Laboratory of Genome Structure and Function, Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan

   Katsunori Fujiki & Katsuhiko Shirahige

4. Laboratory of Chromosome dynamics and genome stability, Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden

   Katsuhiko Shirahige

Contributions

M.H. conceived the study, developed the methodology, and supervised the project. K. Fujiwara, K. Fujiki, and M.H. performed the investigation. K. Fujiki, K.S., I.S., T.N., and M.H. provided resources. M.H. performed formal analysis and visualization, and wrote, edited, and finalized the manuscript. T.O.A. and T.N. contributed to the writing (review and editing).

Corresponding author

Correspondence to Maretoshi Hirai.

Competing interests

The authors declare no competing interests.

Peer review information

Communications Biology thanks Daniel Conway, Zheng Liu, and the other, anonymous, reviewer for their contribution to the peer review of this work. Primary Handling Editors: Yingke Xu & Rosie Bunton-Stasyshyn. A peer review file is available.

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