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Up-regulated α-actin expression is associated with cell adhesion ability in 3-D cultured myocytes subjected to mechanical stimulation

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Abstract

This study was aimed to investigate the alteration of α-actin in three-dimensionally (3-D) cultured myocytes under cyclic tensile stress loading. Myocytes were collected from neonatal SD rat’s lateral pterygoid muscle for primary cell culture. The third-passage cells were implanted and 3-D cultured in poly lactic-co-glycolic acid (PLGA) scaffold, and then subjected to cyclic tensile stress (0.5 Hz, 2,000 μstrain) for 0, 2, 4, 8, 12, and 24 h through a four-point bending strain system. The α-actin mRNA was investigated by semi-quantitative RT–PCR. The α-actin protein expression was examined by immunofluorescent cytochemistry, laser confocal scanning microscopy (LCSM), and image analysis technology. The dynamic adhesion of myocytes to PLGA scaffolds was investigated by fluorescence microscope and the viability of the myocytes was measured by MTT assay. After mechanical loading, the α-actin mRNA increased at 2 h and then declined. The α-actin protein expression kept increased until peaked at 12 h, but declined at 24 h. The time course changing of α-actin protein expression parallelled with that of cell adhesion ability. It is concluded that α-actin expression is probably associated with cell adhesion ability in myocytes subjected to mechanical stimulation.

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References

  1. Lauffenburger DA, Horwitz AF (1996) Cell migration: a physically integrated molecular process. Cell 84:359–369

    Article  CAS  PubMed  Google Scholar 

  2. Viel A (1999) Alpha-actinin and spectrin structures: an unfolding family story. FEBS Lett 460:391–394

    Article  CAS  PubMed  Google Scholar 

  3. Rajfur Z, Roy P, Otey C et al (2002) Dissecting the link between stress fibres and focal adhesions by CALI with EGFP fusion proteins. Nat Cell Biol 4:286–293

    Article  CAS  PubMed  Google Scholar 

  4. Fukami K, Furuhashi K, Inagaki M et al (1992) Requirement of phosphatidylinositol 4,5-bisphosphate for alpha-actinin function. Nature 359:150–152

    Article  CAS  PubMed  Google Scholar 

  5. Clark EA, Brugge JS (1995) Integrins and signal transduction pathways: the road taken. Science 268:233–239

    Article  CAS  PubMed  Google Scholar 

  6. Guvakova MA, Adams JC, Boettiger D (2002) Functional role of alpha-actinin, PI 3-kinase and MEK1/2 in insulin-like growth factor I receptor kinase regulated motility of human breast carcinoma cells. J Cell Sci 115:4149–4165

    Article  CAS  PubMed  Google Scholar 

  7. Otey CA, Carpen O (2004) Alpha-actinin revisited: a fresh look at an old player. Cell Motil Cytoskelet 58:104–111

    Article  CAS  Google Scholar 

  8. Corgan AM, Singleton C, Santoso CB et al (2004) Phosphoinositides differentially regulate alpha-actinin flexibility and function. Biochem J 378:1067–1072

    Article  CAS  PubMed  Google Scholar 

  9. Otey CA, Vasquez GB, Burridge K et al (1993) Mapping of the alpha-actinin binding site within the beta 1 integrin cytoplasmic domain. J Biol Chem 268:21193–21197

    CAS  PubMed  Google Scholar 

  10. Knudsen KA, Soler AP, Johnson KR et al (1995) Interaction of alpha-actinin with the cadherin/catenin cell–cell adhesion complex via alpha-catenin. J Cell Biol 130:67–77

    Article  CAS  PubMed  Google Scholar 

  11. Xu J, Tseng Y, Wirtz D (2000) Strain hardening of actin filament networks. Regulation by the dynamic cross-linking protein alpha-actinin. J Biol Chem 275:35886–35892

    Article  CAS  PubMed  Google Scholar 

  12. Kelly DF, Taylor DW, Bakolitsa C et al (2006) Structure of the alpha-actinin-vinculin head domain complex determined by cryo-electron microscopy. J Mol Biol 357:562–573

    Article  CAS  PubMed  Google Scholar 

  13. Akisaka T, Yoshida H, Inoue S et al (2001) Organization of cytoskeletal F-actin, G-actin, and gelsolin in the adhesion structures in cultured osteoclast. J Bone Miner Res 16:1248–1255

    Article  CAS  PubMed  Google Scholar 

  14. Alenghat FJ, Ingber DE (2002) Mechanotransduction: all signals point to cytoskeleton, matrix, and integrins. Sci STKE 2002:pe6

    Article  PubMed  Google Scholar 

  15. Katsumi A, Orr AW, Tzima E et al (2004) Integrins in mechanotransduction. J Biol Chem 279:12001–12004

    Article  CAS  PubMed  Google Scholar 

  16. Buckwalter JA, Grodzinsky AJ (1999) Loading of healing bone, fibrous tissue, and muscle: implications for orthopaedic practice. J Am Acad Orthop Surg 7:291–299

    CAS  PubMed  Google Scholar 

  17. Yang S, Alnaqeeb M, Simpson H et al (1996) Cloning and characterization of an IGF-1 isoform expressed in skeletal muscle subjected to stretch. J Muscle Res Cell Motil 17:487–495

    Article  CAS  PubMed  Google Scholar 

  18. Yang H, Alnaqeeb M, Simpson H et al (1997) Changes in muscle fibre type, muscle mass and IGF-I gene expression in rabbit skeletal muscle subjected to stretch. J Anat 190(Pt 4):613–622

    Article  PubMed  Google Scholar 

  19. McKoy G, Ashley W, Mander J et al (1999) Expression of insulin growth factor-1 splice variants and structural genes in rabbit skeletal muscle induced by stretch and stimulation. J Physiol 516(Pt 2):583–592

    Article  CAS  PubMed  Google Scholar 

  20. Goldspink G, Williams P, Simpson H (2002) Gene expression in response to muscle stretch. Clin Orthop Relat Res 403:S146–S152

    Article  PubMed  Google Scholar 

  21. Nakanishi Y, Chen G, Komuro H et al (2003) Tissue-engineered urinary bladder wall using PLGA mesh-collagen hybrid scaffolds: a comparison study of collagen sponge and gel as a scaffold. J Pediatr Surg 38:1781–1784

    Article  PubMed  Google Scholar 

  22. Zhu Y, Chan-Park MB, Sin Chian K (2005) The growth improvement of porcine esophageal smooth muscle cells on collagen-grafted poly(DL-lactide-co-glycolide) membrane. J Biomed Mater Res B Appl Biomater 75:193–199

    PubMed  Google Scholar 

  23. Bashur CA, Dahlgren LA, Goldstein AS (2006) Effect of fiber diameter and orientation on fibroblast morphology and proliferation on electrospun poly(D,L-lactic-co-glycolic acid) meshes. Biomaterials 27:5681–5688

    Article  CAS  PubMed  Google Scholar 

  24. Shen H, Hu X, Bei J et al (2008) The immobilization of basic fibroblast growth factor on plasma-treated poly(lactide-co-glycolide). Biomaterials 29:2388–2399

    Article  CAS  PubMed  Google Scholar 

  25. Liu J, Liu T, Zheng Y et al (2006) Early responses of osteoblast-like cells to different mechanical signals through various signaling pathways. Biochem Biophys Res Commun 348:1167–1173

    Article  CAS  PubMed  Google Scholar 

  26. Qi M, Zou S, Han L et al (2009) Expression of bone-related genes in bone marrow MSCs after cyclic mechanical strain: implications for distraction osteogenesis. Int J Oral Sci 1:143–150

    Google Scholar 

  27. Li Y, Zhao Z, Song J et al (2009) Cyclic force upregulates mechano-growth factor and elevates cell proliferation in 3D cultured skeletal myoblasts. Arch Biochem Biophys 490:171–176

    Article  CAS  PubMed  Google Scholar 

  28. Li Y, Song J, Yang P et al (2009) Establishment of a three-dimensional culture and mechanical loading system for skeletal myoblasts. Cell Biol Int 33:192–198

    Article  CAS  PubMed  Google Scholar 

  29. Owan I, Burr DB, Turner CH et al (1997) Mechanotransduction in bone: osteoblasts are more responsive to fluid forces than mechanical strain. Am J Physiol 273:C810–C815

    CAS  PubMed  Google Scholar 

  30. Chandran R, Knobloch TJ, Anghelina M et al (2007) Biomechanical signals upregulate myogenic gene induction in the presence or absence of inflammation. Am J Physiol Cell Physiol 293:C267–C276

    Article  CAS  PubMed  Google Scholar 

  31. Fillinger MF, O’Connor SE, Wagner RJ et al (1993) The effect of endothelial cell coculture on smooth muscle cell proliferation. J Vasc Surg 17:1058–1067 discussion 1067-1058

    Article  CAS  PubMed  Google Scholar 

  32. Mandl EW, van der Veen SW, Verhaar JA et al (2004) Multiplication of human chondrocytes with low seeding densities accelerates cell yield without losing redifferentiation capacity. Tissue Eng 10:109–118

    Article  CAS  PubMed  Google Scholar 

  33. Scadden DT (2006) The stem-cell niche as an entity of action. Nature 441:1075–1079

    Article  CAS  PubMed  Google Scholar 

  34. Roschke AV, Tonon G, Gehlhaus KS et al (2003) Karyotypic complexity of the NCI-60 drug-screening panel. Cancer Res 63:8634–8647

    CAS  PubMed  Google Scholar 

  35. Ross DT, Scherf U, Eisen MB et al (2000) Systematic variation in gene expression patterns in human cancer cell lines. Nat Genet 24:227–235

    Article  CAS  PubMed  Google Scholar 

  36. Irish JM, Hovland R, Krutzik PO et al (2004) Single cell profiling of potentiated phospho-protein networks in cancer cells. Cell 118:217–228

    Article  CAS  PubMed  Google Scholar 

  37. Birgersdotter A, Sandberg R, Ernberg I (2005) Gene expression perturbation in vitro—a growing case for three-dimensional (3D) culture systems. Semin Cancer Biol 15:405–412

    Article  PubMed  Google Scholar 

  38. Butterfield TA, Herzog W (2005) Quantification of muscle fiber strain during in vivo repetitive stretch-shortening cycles. J Appl Physiol 99:593–602

    Article  PubMed  Google Scholar 

  39. Hamill OP, Martinac B (2001) Molecular basis of mechanotransduction in living cells. Physiol Rev 81:685–740

    CAS  PubMed  Google Scholar 

  40. Samaj J, Baluska F, Hirt H (2004) From signal to cell polarity: mitogen-activated protein kinases as sensors and effectors of cytoskeleton dynamicity. J Exp Bot 55:189–198

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  42. Ingber DE (2003) Tensegrity II. How structural networks influence cellular information processing networks. J Cell Sci 116:1397–1408

    Article  CAS  PubMed  Google Scholar 

  43. Hirata H, Tatsumi H, Sokabe M (2008) Mechanical forces facilitate actin polymerization at focal adhesions in a zyxin-dependent manner. J Cell Sci 121:2795–2804

    Article  CAS  PubMed  Google Scholar 

  44. Na S, Collin O, Chowdhury F, Tay B, Ouyang M, Wang Y, Wang N (2008) Rapid signal transduction in living cells is a unique feature of mechanotransduction. Proc Natl Acad Sci USA 105:6626–6631

    Article  CAS  PubMed  Google Scholar 

  45. Wang N, Butler JP, Ingber DE (1993) Mechanotransduction across the cell surface and through the cytoskeleton. Science 260:1124–1127

    Article  CAS  PubMed  Google Scholar 

  46. Zhao X-H, Laschinger C, Arora P et al (2007) Force activates smooth muscle <alpha>-actin promoter activity through the Rho signaling pathway. J Cell Sci 120:1801–1809

    Article  CAS  PubMed  Google Scholar 

  47. Endlich N, Otey CA, Kriz W et al (2007) Movement of stress fibers away from focal adhesions identifies focal adhesions as sites of stress fiber assembly in stationary cells. Cell Motil Cytoskelet 64:966–976

    Article  CAS  Google Scholar 

  48. Hotulainen P, Lappalainen P (2006) Stress fibers are generated by two distinct actin assembly mechanisms in motile cells. J Cell Biol 173:383–394

    Article  CAS  PubMed  Google Scholar 

  49. Schober M, Raghavan S, Nikolova M et al (2007) Focal adhesion kinase modulates tension signaling to control actin and focal adhesion dynamics. J Cell Biol 176:667–680

    Article  CAS  PubMed  Google Scholar 

  50. Yamada KM, Geiger B (1997) Molecular interactions in cell adhesion complexes. Curr Opin Cell Biol 9:76–85

    Article  CAS  PubMed  Google Scholar 

  51. Sims JR, Karp S, Ingber DE (1992) Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape. J Cell Sci 103(Pt 4):1215–1222

    PubMed  Google Scholar 

  52. Liu Jun, Zou Ling, Zheng Yi et al (2007) NF-kB responds to mechanical strains in osteoblast-like cells, and lighter strains create an NF-kB response more readily. Cell Biol Int 31:1220–1224

    Article  CAS  PubMed  Google Scholar 

  53. Grossi A, Yadav K, Lawson MA (2007) Mechanical stimulation increases proliferation, differentiation and protein expression in culture: stimulation effects are substrate dependent. J Biomech 40:3354–3362

    Article  PubMed  Google Scholar 

  54. Kumar A, Murphy R, Robinson P et al (2004) Cyclic mechanical strain inhibits skeletal myogenesis through activation of focal adhesion kinase, Rac-1 GTPase, and NF-kappaB transcription factor. FASEB J 18:1524–1535

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The research was supported by the grants from National Nature Science Foundation of China No. 10472138 and No. 30800212.

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Authors and Affiliations

  1. Department of Orthodontics, State Key Laboratory of Oral Diseases, West China Stomatology Hospital Sichuan University, Chengdu, Sichuan, 610041, People’s Republic of China

    Yu Wang, Zhihe Zhao, Yu Li, Jiapei Wu & Pu Yang

  2. Department of Prosthodontics, College of Stomatology, Chongqing Medical Univesity, Chongqing, 400015, People’s Republic of China

    Youwei Li

  3. Department of Orthodontics, Hainan Stomatology Hospital, Haikou, Hainan, 570125, People’s Republic of China

    Xiaofeng Fan

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  1. Yu Wang
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  2. Zhihe Zhao
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Correspondence to Pu Yang.

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Wang, Y., Zhao, Z., Li, Y. et al. Up-regulated α-actin expression is associated with cell adhesion ability in 3-D cultured myocytes subjected to mechanical stimulation. Mol Cell Biochem 338, 175–181 (2010). https://doi.org/10.1007/s11010-009-0351-7

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  • DOI: https://doi.org/10.1007/s11010-009-0351-7

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