The Number of Cyclic Stretch Regulates Cellular Elasticity in C2C12 Myoblasts

Abstract

Mechanical stimulations have been shown to regulate cellular mechanical properties. However, the stimulation patterns for effective regulation are as yet unclear. We investigated the effects of application of differing numbers of mechanical stimulation sets, each set consisting of 8% extension and compression to cells via deformation of cell culture elastic chamber, on cellular elasticity. Elasticity increased with only a single step-like stretch and with a single step-like stretch after 1 set of mechanical stimulation, whereas elasticity did not change with a single step-like stretch after 10 sets of mechanical stimulation. These results indicate that the increase in cellular elasticity with the single step-like stretch depends on the number of applied mechanical stimulations. Immunofluorescence staining showed that phosphorylation and dephosphorylation of myosin regulatory light chain (MRLC), which regulates intracellular contractile force and cellular elasticity, accompanied cellular elasticity changes. These findings suggest that cellular elasticity changes under cyclic and step-like stretches are mediated by MRLC.

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K. Takemoto, T. Mizutani, K. Tamura, K. Takeda, H. Haga and K. Kawabata, "The Number of Cyclic Stretch Regulates Cellular Elasticity in C2C12 Myoblasts," CellBio, Vol. 1 No. 1, 2012, pp. 1-10. doi: 10.4236/cellbio.2012.11001.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] J. A. Beamish, et al., “Molecular Regulation of Contractile Smooth Muscle Cell Phenotype: Implications for Vascular Tissue Engineering,” Tissue Engineering Part B-Reviews, Vol. 16, No. 5, 2010, pp. 467-491. doi:10.1089/ten.teb.2009.0630
[2] C. Galli, et al., “Osteocytes and WNT: The Mechanical Control of Bone Formation,” Journal of Dental Research, Vol. 89, No. 4, 2010, pp. 331-343. doi:10.1177/0022034510363963
[3] L. Y. Liu, et al., “Mechanisms for Osteogenic Differentiation of Human Mesenchymal Stem Cells Induced by Fluid Shear Stress,” Biomechanics and Modeling in Mechanobiology, Vol. 9, No. 6, 2010, pp. 659-670. doi:10.1007/s10237-010-0206-x
[4] Z. Yin, et al., “Stem Cells for Tendon Tissue Engineering and Regeneration,” Expert Opinion on Biological Therapy, Vol. 10, No. 5, 2010, pp. 689-700. doi:10.1517/14712591003769824
[5] I. Schofield, et al., “Vascular Structural and Functional Changes in Type 2 Diabetes Mellitus—Evidence for the Roles of Abnormal Myogenic Responsiveness and Dyslipidemia,” Circulation, Vol. 16, No. 5, 2002, pp. 30373043. doi:10.1161/01.CIR.0000041432.80615.A5
[6] J. Harle, et al., “Effects of Ultrasound on the Growth and Function of Bone and Periodontal Ligament Cells in Vitro,” Ultrasound in Medicine and Biology, Vol. 27, No. 4, 2001, pp. 579-586. doi:10.1016/S0301-5629(00)00326-4
[7] S. H. Kook, et al., “Cyclic Mechanical Stretch Stimulates the Proliferation of C2C12 Myoblasts and Inhibits Their Differentiation via Prolonged Activation of p38 MAPK,” Molecules and Cells, Vol. 25, No. 4, 2008, pp. 479-486.
[8] K. Naruse, et al., “Involvement of SA Channels in Orienting Response of Cultured Endothelial Cells to Cyclic Stretch,” American Journal of Physiology-Heart and Circulatory Physiology, Vol. 43, No. 5, 1998, pp. H1532-H1538.
[9] T. Mizutani, et al., “Cellular Stiffness Response to External Deformation: Tensional Homeostasis in a Single Fibroblast,” Cell Motility and the Cytoskeleton, Vol. 59, No. 4, 2004, pp. 242-248. doi:10.1002/cm.20037
[10] M. Nagayama, et al., “Contribution of Cellular Contractility to Spatial and Temporal Variations in Cellular Stiffness,” Experimental Cell Research, Vol. 300, No. 2, 2004, pp. 396-405. doi:10.1016/j.yexcr.2004.07.034
[11] K. Tamura, et al., “Visualization of Stretch-Induced Intracellular Tensional Response of Single Fibroblasts by Mechanical Scanning Probe Microscopy,” Japanese Journal of Applied Physics Part 1—Regular Papers Brief Communications & Review Papers, Vol. 46, No. 8B, 2007, pp. 5631-5635.
[12] H. Haga, et al., “Elasticity Mapping of Living Fibroblasts by AFM and Immunofluorescence Observation of the Cytoskeleton,” Ultramicroscopy, Vol. 82, No. 1-4, 2000, pp. 253-258. doi:10.1016/S0304-3991(99)00157-6
[13] S. Na, et al., “Time-Dependent Changes in Smooth Muscle Cell Stiffness and Focal Adhesion Area in Response to Cyclic Equibiaxial Stretch,” Annals of Biomedical Engineering, Vol. 36, No. 3, 2008, pp. 369-380. doi:10.1007/s10439-008-9438-7
[14] T. G. Kuznetsova, et al., “Atomic Force Microscopy Probing of Cell Elasticity,” Micron, Vol. 38, No. 8, 2007, pp. 824-833. doi:10.1016/j.micron.2007.06.011
[15] T. Mizutani, et al., “Diphosphorylation of the Myosin Regulatory Light Chain Enhances the Tension Acting on Stress Fibers in Fibroblasts,” Journal of Cellular Physiology, Vol. 209, No. 3, 2006, pp. 726-731. doi:10.1002/jcp.20773
[16] T. Mizutani, et al., “Regulation of Cellular Contractile Force in Response to Mechanical Stretch by Diphosphorylation of Myosin Regulatory Light Chain via RhoA Signaling Cascade,” Cell Motility and the Cytoskeleton, Vol. 66, No. 7, 2009, pp. 389-397. doi:10.1002/cm.20378
[17] J. T. Stull, et al., “Myosin Light Chain Kinase Phosphorylation in Tracheal Smooth-Muscle,” Journal of Biological Chemistry, Vol. 265, No. 27, 1990, pp. 1668316690.
[18] S. Jungbauer, et al., “Two Characteristic Regimes in Frequency-Dependent Dynamic Reorientation of Fibroblasts on Cyclically Stretched Substrates,” Biophysical Journal, Vol. 95, No. 7, 2008, pp. 3470-3478. doi:10.1529/biophysj.107.128611
[19] K. D. Costa, et al., “Buckling of Actin Stress Fibers: A New Wrinkle in the Cytoskeletal Tapestry,” Cell Motility and the Cytoskeleton, Vol. 52, No. 4, 2002, pp. 266-274. doi:10.1002/cm.10056
[20] P. Reusch, et al., “Mechanical Strain Increases Smooth Muscle and Decreases Nonmuscle Myosin Expression in Rat Vascular Smooth Muscle Cells,” Circulation Research, Vol. 79, No. 5, 1996, pp. 1046-1053. doi:10.1161/01.RES.79.5.1046
[21] D. Kaspar, et al., “Proliferation of Human-Derived Osteoblast-Like Cells Depends on the Cycle Number and Frequency of Uniaxial Strain,” Journal of Biomechanics, Vol. 35, No. 7, 2002, pp. 873-880. doi:10.1016/S0021-9290(02)00058-1
[22] K. Jani and F. Schock, “Molecular Mechanisms of Mechanosensing in Muscle Development,” Developmental Dynamics, Vol. 238, No. 6, 2009, pp. 1526-1534. doi:10.1002/dvdy.21972
[23] A. J. Ricci, et al., “Mechano-Electrical Transduction: New Insights into Old Ideas,” Journal of Membrane Biology, Vol. 209, No. 2-3, 2006, pp. 71-88. doi:10.1007/s00232-005-0834-8
[24] P. G. Smith, et al., “Mechanical Stress Increases RhoA Activation in Airway Smooth Muscle Cells,” American Journal of Respiratory Cell and Molecular Biology, Vol. 28, No. 4, 2003, pp. 436-442. doi:10.1165/rcmb.4754
[25] J. J. Cunningham, et al., “Externally Applied Cyclic Strain Regulates Localization of Focal Contact Components in Cultured Smooth Muscle Cells,” Annals of Biomedical Engineering, Vol. 30, No. 7, 2002, pp. 927935. doi:10.1114/1.1500408
[26] S. S. An and C. M. Hai, “Mechanical Signals and Mechanosensitive Modulation of Intracellular [Ca2+] in Smooth Muscle,” American Journal of Physiology-Cell Physiology, Vol. 279, No. 5, 2000, pp. C1375-C1384.
[27] S. Komatsu, et al., “Effects of the Regulatory Light Chain Phosphorylation of Myosin II on Mitosis and Cytokinesis of Mammalian Cells,” Journal of Biological Chemistry, Vol. 275, No. 44, 2000, pp. 34512-34520. doi:10.1074/jbc.M003019200
[28] M. F. Santos, et al., “Rho Proteins Play a Critical Role in Cell Migration during the Early Phase of Mucosal Restitution,” Journal of Clinical Investigation, Vol. 100, No. 1, 1997, pp. 216-225. doi:10.1172/JCI119515

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