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Relationship between Apical Membrane Elasticity and Stress Fiber Organization in Fibroblasts Analyzed by Fluorescence and Atomic Force Microscopy

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Abstract

To investigate the relationship between cellular microelasticity and the structural features of cytoskeletons (CSKs), a microindentation test for apical cell membranes and observation of the spatio-distribution of actin CSKs of fibroblasts were performed by fluorescence and atomic force microscopy (FM/AFM). The indentation depths of apical cell membranes were measured from AFM force–indentation (f–i) curves under equal final loads and mapped two-dimensionally to show the relative distribution of local microelasticity on cell membranes. Intracellular spatial distribution of actin CSKs was visualized fluorescently by high Z-resolution cross-sectional observation of a cell on which indentation mapping analysis had been performed in advance. Structural features of stress fibers (SFs) were observed as three typical patterns of dense SF, sparse SF and sparser SF cell groups, which were quantitated using the degree of orientation in apical SFs (ASFs) that had been defined using two-dimensional Fourier analysis. In indentation depth maps, the upper nuclear region was markedly softer than the pseudopodium region. The mean indentation depth of the upper nuclear region decreased with increased SF density in whole cells and the degree of orientation of ASF, although the pseudopodium region did not exhibit such a trend. The apical membrane of adhered cells was found to tend to stiffen with the increase in both density and degree of orientation of SFs.

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References

  • Byers HR, White GE, Fujiwara K (1984) Organization and function of stress fibers in cells in vitro and in situ. Cell Muscle Motil 5:83–137

    Google Scholar 

  • Chang L, Kious T, Yorgancioglu M, Keller D, Pfeiffer J (1993) Cytoskeleton of living, unstained cells imaged by scanning force microscopy. Biophys J 64:1282–1286

    Google Scholar 

  • Dembo M, Wang YL (1999) Stresses at the cell-to-substrate interface during locomotion of fibroblasts. Biophys J 76:2307–2316

    Google Scholar 

  • Evans E, Yeung A (1989) Apparent viscosity and cortical tension of blood granulocytes determined by micropipette aspiration. Biophys J 56:139–149

    Google Scholar 

  • Folkman J, Moscona A (1978) Role of cell shape in growth control. Nature 273:345–349

    Article  Google Scholar 

  • Hassan EA, Heinz WF, Antonik MD, D’Costa, NP, Nageswaran S, Schoenenberger CA, Hoh J (1998) Relative microelastic mapping of living cells by atomic force microscopy. Biophys J 74:1564–1578

    Google Scholar 

  • Henderson E, Haydon PG, Sakaguchi DS (1992) Actin filament dynamics in living glial cells imaged by atomic force microscopy. Science 257:1944–1946

    Article  Google Scholar 

  • Hertz H (1881) Ueber die Beruhrrung fester elastischer Korper. J Reine Angew Math 92:156–171

    Google Scholar 

  • Hoh JH, Schoenenberger CA (1994) Surface morphology and mechanical properties of MDCK monolayers by atomic force microscopy. J Cell Sci 107:1105–1114

    Google Scholar 

  • Ingber DE (2003) Tensegrity I. Cell structure and hierarchical systems biology. J Cell Sci 116:1157–1173

    Google Scholar 

  • Ingber DE, Folkman J (1989) Tension and compression as basic determinants of cell form and function:utilization of a cellular tensegrity mechanism. In: Stein WD, Bronner F (eds.) Cell shape:determinants, regulation, and regulatory role. Academic Inc., San Diego

    Google Scholar 

  • Kasas S, Gotzos V, Celio MR (1993) Observation of living cells using the atomic force microscope. Biophys J 64:539–544

    Google Scholar 

  • Katoh K, Masuda M, Kano Y, Jinguji Y, Fujiwara K (1995) Focal adhesion proteins associated with apical stress fibers of human fibroblasts. Cell Motil Cytoskelet 31:177–195

    Article  Google Scholar 

  • Katoh K, Kano Y, Masuda M, Fujiwara K (1996) Mutually exclusive distribution of the focal adhesion associated proteins and the erythrocyte membrane skeleton proteins in the human fibroblast plasma membrane undercoat. Cell Struct Funct 21:27–39

    Article  Google Scholar 

  • Lo CM, Wang HB, Dembo M, Wang YL (2000) Cell movement is guided by the rigidity of the substrate. Biophys J 79:144–152

    Google Scholar 

  • von der Mark K, Gauss V, von der Mark H, Muller P (1977) Relationship between cell shape and type of collagen synthesized as chondrocytes lose their cartilage phenotype in culture. Nature 267:531–532

    Article  Google Scholar 

  • Munevar S, Wang YL, Dembo M (2001) Traction force microscopy of migrating normal and H-ras transformed 3T3 fibroblasts. Biophys J 80:1744–1757

    Article  Google Scholar 

  • Nagayama M, Haga H, Kawabata K (2001) Drastic change of local stiffness distribution correlating to cell migration in living fibroblasts. Cell Motil Cytoskelet 50:173–179

    Article  Google Scholar 

  • Osborn M, Born T, Koitsch HJ, Weber K (1978) Stereo immunofluorescence microscopy:I Three-dimensional arrangement of microfilaments, microtubules and tonofilaments. Cell 14:477–488

    Article  Google Scholar 

  • Oster G (1989) Cell motility and tissue morphogenesis. In: Stein WD, Bronner F (eds) Cell shape:determinants, regulation, and regulatory role. Academic, San Diego

    Google Scholar 

  • Petersen NO, McConnaughey WB, Elson EL (1982) Dependence of locally measured cellular deformability on position on the cell, temperature, and cytochalasin B. Proc Natl Acad Sci USA 79:5327–5331

    Article  Google Scholar 

  • Pourdeyhimi B (1997) Measuring fiber orientation in nonwovens Part III:Fourier transform. Text Res J 67:143–151

    Google Scholar 

  • Radmacher M, Fritz M, Kacher CM, Cleveland JP, Hansma PK (1996) Measuring the viscoelastic properties of human platelets with the atomic force microscope. Biophys J 70:556–567

    Google Scholar 

  • Rotsch C, Radmacher M (2000) Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts:atomic force microscopy study. Biophys J 78:520–535

    Google Scholar 

  • Shroff SG, Saner DR, Lal R (1995) Dynamic micromechanical properties of cultured rat arterial myocytes measured by atomic force microscopy. Am Physiol Soc 269:C286-C292

    Google Scholar 

  • Sneddon IN (1965) The relation between load and penetration in the axisymmetric Boussinesq problem for a punch of arbitrary profile. Int J Eng Sci 3:47–57

    Article  MATH  MathSciNet  Google Scholar 

  • Sokabe M, Naruse K, Nunogaki K (1997) Mechanosensitive ion channels:single channel vs. whole cell activities. Prog Cell Res 6:139–149

    Google Scholar 

  • Stein WD, Bronner F (eds) (1989) Cell shape:determinants, regulation, and regulatory role, Chapt 8–11. Academic, San Diego

    Google Scholar 

  • Wang N, Ingber DE (1995) Probing transmembrane mechanical coupling and cytomechanics using magnetic twisting cytometry. Biochem Cell Biol 73:1–9

    Article  MATH  Google Scholar 

  • Wang N, Bulter JP, Ingber DE (1993) Mechanotransduction across the cell surface and through the cytoskeleton. Science 20:1124–1127

    Article  Google Scholar 

  • Wang N, Naruse K, Stamenovic D, Fredberg J, Mijailovich SM, Tolic-Norrelykke IM, Polte T, Mannix R, Ingber DE (2001a) Mechanical behavior in living cells consistent with the tensegrity model. Proc Natl Acad Sci USA 98:7765–7770

    Article  Google Scholar 

  • Wang HB, Dembo M, Hanks SK, Wang YL (2001b) Focal adhesion kinase is involved in mechanosensing during fibroblast migration. Proc Natl Acad Sci USA 98:11295–11300

    Article  Google Scholar 

  • Wang GN, Tolic-noprelykke IM, Chen J, Mijailovich SM, Butler JP, Fredberg JJ, Stamenovic D (2002) Cell prestress I:stiffness and prestress are closely associated adherent contractile cells. Am J Physiol Cell Physiol 282:C606-C612

    Google Scholar 

  • White G, Fujiwara K (1986) Expression and intracellular distribution of stress fibers in aortic endothelium. J Cell Biol 103:63–70

    Article  Google Scholar 

  • Wood EJ (1990) Applying Fourier and associated transforms to pattern characterization in textiles. Text Res J 60:212–220

    Google Scholar 

  • Yoshinaga N, Yoshikawa K, Kidoaki S (2002) Multi-scaling in a long semi-flexible polymer chain in 2D. J Chem Phys 116:9926–9929

    Article  Google Scholar 

  • Zhu C, Bao G, Wang N (2000) Cell mechanics:mechanical response, cell adhesion, and molecular deformation. Annu Rev Biomed Eng 2:189–226

    Article  MATH  Google Scholar 

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Kidoaki, S., Matsuda, T. & Yoshikawa, K. Relationship between Apical Membrane Elasticity and Stress Fiber Organization in Fibroblasts Analyzed by Fluorescence and Atomic Force Microscopy. Biomech Model Mechanobiol 5, 263–272 (2006). https://doi.org/10.1007/s10237-006-0048-8

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  • DOI: https://doi.org/10.1007/s10237-006-0048-8

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