Abstract
Actin stress fibers (SFs) play important roles in cellular mechanotransduction and in regulation of various cellular functions. Stress fibers generate internal tension and contribute to physical interactions between cells and extracellular matrices. We recently found that SFs in vascular smooth muscle cells (SMCs) cultured on a two-dimensional substrate mechanically interact with cell nucleus via nuclear membrane proteins, and that the internal tension of SFs is transmitted directly to the nucleus. However, SFs exist on both the apical side and the basal side of adherent cells on a substrate, and it remains unclear whether these two types of SFs play different roles on the mechanical environment around the nucleus. Here, we investigated differences between the apical and basal stress fibers (BSFs) in SMCs by using a laser nano-scissor technique. We microdissected apical SFs running across the top surface of nucleus (actin cap fibers: ACFs) or BSFs underneath the nucleus by using a laboratory-built laser nano-scissor and observed the subsequent mechanical responses of the SFs and the nucleus. Shortening of the dissected fibers was significantly greater in the ACFs than in the BSFs. Nuclei also moved in the direction of retraction of the dissected fibers, and displacement and local deformation of the nucleus was more remarkable after the dissection of the ACFs than after that of the BSFs. ACFs mostly aligned in the major axis of the nucleus, whereas BSFs showed a weak alignment with asymmetry: the direction of BSFs was rotated clockwise by ~10° from the major axis of the nucleus. These results indicate that ACFs and BSFs play different roles in mechanical regulation of the nucleus, and that intracellular tension is transmitted to the nucleus more efficiently by ACFs. ACFs may play significant roles in controlling the intranuclear distribution of DNA through intracellular orientation and positioning of the nucleus.
Similar content being viewed by others
References
Anno, T., N. Sakamoto, and M. Sato. Role of nesprin-1 in nuclear deformation in endothelial cells under static and uniaxial stretching conditions. Biochem. Biophys. Res. Commun. 424(1):94–99, 2012.
Chen, C. S., M. Mrksich, S. Huang, G. M. Whitesides, and D. E. Ingber. Geometric control of cell life and death. Science 276:1425–1428, 1997.
Collinsworth, A. M., S. Zhang, W. E. Kraus, and G. A. Truskey. Apparent elastic modulus and hysteresis of skeletal muscle cells throughout differentiation. Am. J. Physiol. Cell Physiol. 283:1219–1227, 2002.
Colombelli, J., A. Besser, H. Kress, E. G. Reynaud, P. Girard, E. Caussinus, U. Haselmann, J. V. Small, U. S. Schwarz, and E. H. Stelzer. Mechanosensing in actin stress fibers revealed by a close correlation between force and protein localization. J. Cell Sci. 122(Pt 10):1665–1679, 2009.
Gerlitz, G., and M. Bustin. The role of chromatin structure in cell migration. Trends Cell Biol. 21(1):6–11, 2011.
Ingber, D. E. Fibronectin controls capillary endothelial cell growth by modulating cell shape. Proc. Natl Acad. Sci. U.S.A. 87:3579–3583, 1990.
Khatau, S. B., R. J. Bloom, S. Bajpai, D. Razafsky, S. Zang, A. Giri, P. H. Wu, J. Marchand, A. Celedon, C. M. Hale, S. X. Sun, D. Hodzic, and D. Wirtz. The distinct roles of the nucleus and nucleus–cytoskeleton connections in three-dimensional cell migration. Sci. Rep. 2:488, 2012. doi:10.1038/srep00488.
Khatau, S. B., C. M. Hale, P. J. Stewart-Hutchinson, M. S. Patel, C. L. Stewart, P. C. Searson, D. Hodzic, and D. Wirtz. A perinuclear actin cap regulates nuclear shape. Proc. Natl Acad. Sci. U.S.A. 106(45):19017–19022, 2009.
Kim, D. H., S. B. Khatau, Y. Feng, S. Walcott, S. X. Sun, G. D. Longmore, and D. Wirtz. Actin cap associated focal adhesions and their distinct role in cellular mechanosensing. Sci. Rep. 2:555, 2012. doi:10.1038/srep00555.
King, M., T. Drivas, and G. Blobel. A network of nuclear envelope membrane proteins linking centromeres to microtubules. Cell 134:427–438, 2008.
Kumar, S., I. Z. Maxwell, A. Heisterkamp, T. R. Polte, T. P. Lele, M. Salanga, E. Mazur, and D. E. Ingber. Viscoelastic retraction of single living stress fibers and its impact on cell shape, cytoskeletal organization, and extracellular matrix mechanics. Biophys. J. 90(10):3762–3773, 2006.
Luxton, G. W., E. R. Gomes, E. S. Folker, E. Vintinner, and G. G. Gundersen. Linear arrays of nuclear envelope proteins harness retrograde actin flow for nuclear movement. Science 329(5994):956–959, 2010.
Minc, N., D. Burgess, and F. Chang. Influence of cell geometry on division-plane positioning. Cell 144(3):414–426, 2011.
Nagayama, K., Y. Kimura, N. Makino, and T. Matsumoto. Strain waveform dependence of stress fiber reorientation in cyclically stretched osteoblastic cells: effects of viscoelastic compression of stress fibers. Am. J. Physiol. Cell Physiol. 302:1469–1478, 2012.
Nagayama, K., and T. Matsumoto. Contribution of actin filaments and microtubules to quasi-in situ tensile properties and internal force balance of cultured smooth muscle cells on a substrate. Am. J. Physiol. Cell Physiol. 295:1569–1578, 2008.
Nagayama, K., and T. Matsumoto. Estimation of single stress fiber stiffness in cultured aortic smooth muscle cells under relaxed and contracted states: its relation to dynamic rearrangement of stress fibers. J. Biomech. 43:1443–1449, 2010.
Nagayama, K., and T. Matsumoto. Dynamic change in morphology and traction forces at focal adhesions in cultured vascular smooth muscle cells during contraction. Cell. Mol. Bioeng. 4(3):348–357, 2011.
Nagayama, K., Y. Yahiro, and T. Matsumoto. Stress fibers stabilize the position of intranuclear DNA through mechanical connection with the nucleus in vascular smooth muscle cells. FEBS Lett. 585(24):3992–3997, 2011.
Orr, A. W., B. P. Helmke, B. R. Blackman, and M. A. Schwartz. Mechanisms of mechanotransduction. Dev. Cell 10:11–20, 2006.
Russell, R. J., S. L. Xia, R. B. Dickinson, and T. P. Lele. Sarcomere mechanics in capillary endothelial cells. Biophys. J. 97(6):1578–1585, 2009.
Smilenov, L. B., A. Mikhailov, R. J. Pelham, E. E. Marcantonio, and G. G. Gundersen. Focal adhesion motility revealed in stationary fibroblasts. Science 286(5442):1172–1174, 1999.
Smith, P. G., C. Roy, S. Fisher, Q. Q. Huang, and F. Brozovich. Selected contribution: mechanical strain increases force production and calcium sensitivity in cultured airway smooth muscle cells. J. Appl. Physiol. 89(5):2092–2098, 2000.
Tanner, K., A. Boudreau, M. J. Bissell, and S. Kumar. Dissecting regional variations in stress fiber mechanics in living cells with laser nanosurgery. Biophys. J. 99(9):2775–2783, 2010.
Vogel, V., and M. Sheetz. Local force and geometry sensing regulate cell functions. Nat. Rev. Mol. Cell Biol. 7:265–275, 2006.
Wang, N., J. Tytell, and D. Ingber. Mechanotransduction at a distance: mechanically coupling the extracellular matrix with the nucleus. Nat. Rev. Mol. Cell. Biol. 10:75–82, 2009.
Wu, J., R. B. Dickinson, and T. P. Lele. Investigation of in vivo microtubule and stress fiber mechanics with laser ablation. Integr. Biol. (Camb.) 4(5):471–479, 2012.
Xiong, H., F. Rivero, U. Euteneuer, S. Mondal, S. Mana-Capelli, D. Larochelle, A. Vogel, B. Gassen, and A. A. Noegel. Dictyostelium Sun-1 connects the centrosome to chromatin and ensures genome stability. Traffic 9(5):708–724, 2008.
Acknowledgments
This work was supported in part by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (nos. 24680051, 24650257, and 25111711 to K.N., and nos. 22127008 and 22240055 to T.M.), and the Hibi Science Foundation, Japan (K.N.).
Conflicts of interest
The authors declare that they have no conflict of interest with regards to this manuscript.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Associate Editor David Sept oversaw the review of this article.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Nagayama, K., Yahiro, Y. & Matsumoto, T. Apical and Basal Stress Fibers have Different Roles in Mechanical Regulation of the Nucleus in Smooth Muscle Cells Cultured on a Substrate. Cel. Mol. Bioeng. 6, 473–481 (2013). https://doi.org/10.1007/s12195-013-0294-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12195-013-0294-7