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Nanotechnology in Mechanobiology: Mechanical Manipulation of Cells and Organelle While Monitoring Intracellular Signaling

  • Hitoshi Tatsumi
  • Kimihide Hayakawa
  • Masahiro Sokabe

Abstract

Cell migration requires a regulated interplay of actin-filament dynamics and turnover of cell-matrix adhesions. These processes are mechanically coupled by a large complex of cytoplasmic proteins linking adhesive molecules to cytoskeletons. To explore the molecular mechanisms underlying the force-dependent cell responses including remodeling of cytoskeletons, focal adhesive contacts, and of cell-shapes, it is crucial to apply precisely controlled mechanical stimuli (ranging from pN to nN) to cells and to observe the response with a high spatial–temporal resolution (ranging from 100 nm to 1 μm, and 1 ms to 1 min) in living cells. In this chapter, we describe a variety of methods for controlled mechanical stimulations and for monitoring the responses of cells, and our recent research. We employed precisely controlled mechanical stimuli to explore the molecular mechanism underlying the mechanosensing, and the mechano-induced signaling, e.g., activation of mechanosensitive (MS) channels, intracellular calcium ion concentration ([Ca2+]i) increases, adhesive contact formation and mechano-induced reorganization of cytoskeletons in live cells including neuronal growth cones and human umbilical vein endothelial cells (HUVECs).

Keywords

Actin Cytoskeleton Dorsal Root Ganglion Neuron Growth Cone Actin Stress Fiber Stretch Axis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was supported in part by Grants-in-aid for General Scientific Research (#13480216 to M.S. and #14580769 to H.T.), Scientific Research on Priority Areas (#15086270 to M.S.) and Creative Research (#16GS0308 to M.S.) from the Ministry of Education Science Sports and Culture and a grant from Japan Space Forum (to M.S. and H.T.).

References

  1. Block SM (1990) Optical tweezers: a new tool for biophysics. In: Noninvasive techniques in cell biology. Wiley-Liss Inc, New York, pp 375–402Google Scholar
  2. Byfield FJ, Aranda-Espinoza H, Romanenko VG, Rothblat GH, Levitan I (2004) Cholesterol depletion increases membrane stiffness of aortic endothelial cells. Biophys J 87  :  3336–3343PubMedCrossRefGoogle Scholar
  3. Corey DP, Garcia-Anoveros J, Holt JR, Kwan KY, Lin SY, Vollrath MA, Amalfitano A, Cheung ELM, Derfler BH, Duggan A, Geleoc GSG, Gray PA, Hoffman MP, Rehm HL, Tamasauskas D, Zhang DS (2004) TRPA1 is a candidate for the mechanosensitive transduction channel of vertebrate hair cells. Nature 432  :  723–730PubMedCrossRefGoogle Scholar
  4. Corry B, Rigby P, Liu ZW, Martinac B (2005) Conformational changes involved in MscL channel gating measured using FRET spectroscopy. Biophys J 89  :  L49–L51PubMedCrossRefGoogle Scholar
  5. Dai J, Sheetz MP (1995) Mechanical properties of neuronal growth cone membranes studied by tether formation with laser optical tweezers. Biophys J 68  :  988–996PubMedCrossRefGoogle Scholar
  6. Dai J, Sheetz MP (1999) Membrane tether formation from blebbing cells. Biophys J 77  :  3363–3370PubMedCrossRefGoogle Scholar
  7. Dembo M, Wang YL (1999) Stresses at the cell-to-substrate interface during locomotion of fibroblasts. Biophys J 76  :  2307–2316PubMedCrossRefGoogle Scholar
  8. Doyle AD, Lee J (2005) Cyclic changes in keratocyte speed and traction stress arise from Ca2+-dependent regulation of cell adhesiveness. J Cell Sci 118  :  369–379PubMedCrossRefGoogle Scholar
  9. Drake CJ, Davis LA, Hungerford JE, Little CD (1992) Perturbation of beta 1 integrin-mediated adhesions results in altered somite cell shape and behavior. Dev Biol 149  :  327–338PubMedCrossRefGoogle Scholar
  10. Du HP, Gu GQ, William CM, Chalfie M (1996) Extracellular proteins needed for C-elegans mechanosensation. Neuron 16  :  183–194PubMedCrossRefGoogle Scholar
  11. Evans EA, Waugh R (1977) Osmotic correction to elastic area compressibility measurements on red-cell membrane. Biophys J 20  :  307–313PubMedCrossRefGoogle Scholar
  12. Fukushige T, Siddiqui ZK, Chou M, Culotti JG, Gogonea CB, Siddiqui SS, Hamelin M (1999) MEC-12, an alpha-tubulin required for touch sensitivity in C-elegans. J Cell Sci 112  :  395–403PubMedGoogle Scholar
  13. Giannone G, Jiang G, Sutton DH, Critchley DR, Sheetz MP (2003) Talin1 is critical for force-dependent reinforcement of initial integrin-cytoskeleton bonds but not tyrosine kinase activation. J Cell Biol 163  :  409–419PubMedCrossRefGoogle Scholar
  14. Gottlieb P, Folgering J, Maroto R, Raso A, Wood TG, Kurosky A, Bowman C, Bichet D, Patel A, Sachs F, Martinac B, Hamill OP, Honore E (2008) Revisiting TRPC1 and TRPC6 mechanosensitivity. Pflugers Archiv Eur J Physiol 455  :  1097–1103CrossRefGoogle Scholar
  15. Gullingsrud J, Schulten K (2003) Gating of MscL studied by steered molecular dynamics. Biophys J 85  :  2087–2099PubMedCrossRefGoogle Scholar
  16. Hayakawa K, Tatsumi H, Sokabe M (2008) Actin stress fibers transmit and focus force to activate mechanosensitive channels. J Cell Sci 121  :  496–503PubMedCrossRefGoogle Scholar
  17. Hirata H, Tatsumi H, Sokabe M (2004) Tension-dependent formation of stress fibers in fibroblasts: a study using semi-intact cells. JSME Int J Ser C 47  :  962–969CrossRefGoogle Scholar
  18. Hodgkin AL, Keynes RD (1957) Movements of labelled calcium in squid giant axons. J Physiol (Lond) 138  :  253–281Google Scholar
  19. Hu SH, Chen JX, Fabry B, Numaguchi Y, Gouldstone A, Ingber DE, Fredberg JJ, Butler JP, Wang N (2003) Intracellular stress tomography reveals stress focusing and structural anisotropy in cytoskeleton of living cells. Am J Physiol Cell Physiol 285  :  C1082–C1090PubMedGoogle Scholar
  20. Iba T, Sumpio BE (1991) Morphological response of human endothelial cells subjected to cyclic strain in vitro. Microvasc Res 42  :  245–254PubMedCrossRefGoogle Scholar
  21. Imai K, Tatsumi H, Katayama Y (2000) Mechanosensitive chloride channels on the growth cones of cultured rat dorsal root ganglion neurons. Neuroscience 97  :  347–355PubMedCrossRefGoogle Scholar
  22. Jacobson BS, Cronin J, Branton D (1978) Coupling polylysine to glass beads for plasma membrane isolation. Biochim Biophys Acta 506  :  81–96PubMedCrossRefGoogle Scholar
  23. Kawakami K, Tatsumi H, Sokabe M (2001) Dynamics of integrin clustering at focal contacts of endothelial cells studied by multimode imaging microscopy. J Cell Sci 114  :  3125–3135PubMedGoogle Scholar
  24. Kojima H, Ishijima A, Yanagida T (1994) Direct measurement of stiffness of single actin filaments with and without tropomyosin by in vitro nanomanipulation. Proc Natl Acad Sci USA 91  :  12962–12966PubMedCrossRefGoogle Scholar
  25. Lansman JB, Hallam TJ, Rink TJ (1987) Single stretch-activated ion channels in vascular endothelial cells as mechanotransducers? Nature 325  :  811–813PubMedCrossRefGoogle Scholar
  26. Machiyama H, Tatsumi H, Sokabe M (2009) Structural changes in the cytoplasmic domain of the mechanosensitive channel MscS during opening. Biophys J 97  :  1048–1057PubMedCrossRefGoogle Scholar
  27. Maroto R, Raso A, Wood TG, Kurosky A, Martinac B, Hamill OP (2005) TRPC1 forms the stretch-activated cation channel in vertebrate cells. Nat Cell Biol 7  :  179–185PubMedCrossRefGoogle Scholar
  28. Munevar S, Wang Y, Dembo M (2001) Traction force microscopy of migrating normal and H-ras transformed 3T3 fibroblasts. Biophys J 80  :  1744–1757PubMedCrossRefGoogle Scholar
  29. Munevar S, Wang YL, Dembo M (2004) Regulation of mechanical interactions between fibroblasts and the substratum by stretch-activated Ca2+ entry. J Cell Sci 117  :  85–92PubMedCrossRefGoogle Scholar
  30. Naruse K, Sokabe M (1993) Involvement of stretch-activated ion channels in Ca2+ mobilization to mechanical stretch in endothelial cells. Am J Physiol 264  :  C1037–C1044PubMedGoogle Scholar
  31. Naruse K, Sai X, Yokoyama N, Sokabe M (1998a) Uni-axial cyclic stretch induces c-src activation and translocation in human endothelial cells via SA channel activation. FEBS Lett 441  :  111–115PubMedCrossRefGoogle Scholar
  32. Naruse K, Yamada T, Sokabe M (1998b) Involvement of SA channels in orienting response of cultured endothelial cells to cyclic stretch. Am J Physiol 274  :  H1532–H1538PubMedGoogle Scholar
  33. Pollard TD, Borisy GG (2003) Cellular motility driven by assembly and disassembly of actin filaments. Cell 112  :  453–465PubMedCrossRefGoogle Scholar
  34. Popp R, Hoyer J, Meyer J, Galla HJ, Gogelein H (1992) Stretch-activated non-selective cation channels in the antiluminal membrane of porcine cerebral capillaries. J Physiol 454  :  435–449PubMedGoogle Scholar
  35. Pourati J, Maniotis A, Spiegel D, Schaffer JL, Butler JP, Fredberg JJ, Ingber DE, Stamenovic D, Wang N (1998) Is cytoskeletal tension a major determinant of cell deformability in adherent endothelial cells? Am J Physiol 274  :  C1283–C1289PubMedGoogle Scholar
  36. Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, Parsons JT, Horwitz AR (2003) Cell migration: integrating signals from front to back. Science 302  :  1704–1709PubMedCrossRefGoogle Scholar
  37. Sachs F (1991) Mechanical transduction by membrane ion channels: a mini review. Mol Cell Biochem 104  :  57–60PubMedCrossRefGoogle Scholar
  38. Shi F, Chiu YJ, Cho YS, Bullard TA, Sokabe M, Fujiwara K (2007) Down-regulation of ERK but not MEK phosphorylation in cultured endothelial cells by repeated changes in cyclic stretch. Cardiovasc Res 73  :  813–822PubMedCrossRefGoogle Scholar
  39. Shirinsky VP, Antonov AS, Birukov KG, Sobolevsky AV, Romanov YA, Kabaeva NV, Antonova GN, Smirnov VN (1989) Mechano-chemical control of human endothelium orientation and size. J Cell Biol 109  :  331–339PubMedCrossRefGoogle Scholar
  40. Sokabe M, Sachs F, Jing Z (1991) Quantitative video microscopy of patch clamped membranes stress, strain, capacitance, and stretch channel activation. Biophys J 59  :  722–728PubMedCrossRefGoogle Scholar
  41. Sukharev SI, Sigurdson WJ, Kung C, Sachs F (1999) Energetic and spatial parameters for gating of the bacterial large conductance mechanosensitive channel, MscL. J Gen Physiol 113  :  525–540PubMedCrossRefGoogle Scholar
  42. Szaszak M, Gaborik Z, Turu G, McPherson PS, Clark AJ, Catt KJ, Hunyady L (2002) Role of the proline-rich domain of dynamin-2 and its interactions with Src homology 3 domains during endocytosis of the AT1 angiotensin receptor. J Biol Chem 277  :  21650–21656PubMedCrossRefGoogle Scholar
  43. Tanaka K, Naruse K, Sokabe M (2005) Effects of mechanical stresses on the migrating behavior of endothelial cells. In: Wada H (ed) Biomechanics at micro and nanoscale levels, vol I. World Scientific Publishing, Singapore, pp 75–87CrossRefGoogle Scholar
  44. Tatsumi H, Katayama Y (1999) Growth cones exhibit enhanced cell–cell adhesion after neurotransmitter release. Neuroscience 99  :  855–865CrossRefGoogle Scholar
  45. Wang GX, Poo MM (2005) Requirement of TRPC channels in netrin-1-induced chemotropic turning of nerve growth cones. Nature 434  :  898–904PubMedCrossRefGoogle Scholar
  46. Wang JG, Miyazu M, Matsushita E, Sokabe M, Naruse K (2001) Uniaxial cyclic stretch induces focal adhesion kinase (FAK) tyrosine phosphorylation followed by mitogen-activated protein kinase (MAPK) activation. Biochem Biophys Res Commun 288  :  356–361PubMedCrossRefGoogle Scholar
  47. Yao X, Kwan H, Huang Y (2001) Stretch-sensitive switching among different channel sublevels of an endothelial cation channel. Biochim Biophys Acta 1511  :  381–390PubMedCrossRefGoogle Scholar
  48. Zamir E, Katz BZ, Aota S, Yamada KM, Geiger B, Kam Z (1999) Molecular diversity of cell-matrix adhesions. J Cell Sci 112(Pt 11)  :  1655–1669PubMedGoogle Scholar
  49. Zenisek D, Davila V, Wan L, Almers W (2003) Imaging calcium entry sites and ribbon structures in two presynaptic cells. J Neurosci 23  :  2538–2548PubMedGoogle Scholar
  50. Zhang Y, Gao F, Popov VL, Wen JW, Hamill OP (2000) Mechanically gated channel activity in cytoskeleton-deficient plasma membrane blebs and vesicles from Xenopus oocytes. J Physiol (Lond) 523  :  117–130CrossRefGoogle Scholar
  51. Zou H, Lifshitz LM, Tuft RA, Fogarty KE, Singer JJ (2002) Visualization of Ca2+ entry through single stretch-activated cation channels. Proc Natl Acad Sci USA 99  :  6404–6409PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2011

Authors and Affiliations

  • Hitoshi Tatsumi
    • 1
  • Kimihide Hayakawa
    • 2
  • Masahiro Sokabe
    • 3
  1. 1.Department of PhysiologyNagoya University Graduate School of MedicineNagoyaJapan
  2. 2.ICORP/ORST, Cell Mechanosensing Project, Japan Science and Technology AgencyNagoyaJapan
  3. 3.ICORP/ORST, Cell Mechanosensing Project, Japan Science and Technology AgencyNagoyaJapan

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