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Quantitative Studies of Endothelial Cell Fibronectin and Filamentous Actin (F-Actin) Coalignment in Response to Shear Stress

Published online by Cambridge University Press:  12 September 2017

Xianghui Gong*
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Xixi Zhao
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Bin Li
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Yan Sun
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Meili Liu
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Yan Huang
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Xiaoling Jia
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Jing Ji
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China
Yubo Fan*
Affiliation:
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, People’s Republic of China National Research Center for Rehabilitation Technical Aids, Beijing 100176, People’s Republic of China
*
*Corresponding authors. xhgong@buaa.edu.cn; yubofan@buaa.edu.cn
*Corresponding authors. xhgong@buaa.edu.cn; yubofan@buaa.edu.cn
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Abstract

Both fibronectin (FN) and filamentous actin (F-actin) fibers play a critical role for endothelial cells (ECs) in responding to shear stress and modulating cell alignment and functions. FN is dynamically coupled to the F-actin cytoskeleton via focal adhesions. However, it is unclear how ECs cooperatively remodel their subcellular FN matrix and intracellular F-actin cytoskeleton in response to shear stress. Current studies are hampered by the lack of a reliable and sensitive quantification method of FN orientation. In this study, we developed a MATLAB-based feature enhancement method to quantify FN and F-actin orientation. The role of F-actin in FN remodeling was also studied by treating ECs with cytochalasin D. We have demonstrated that FN and F-actin codistributed and coaligned parallel to the flow direction, and that F-actin alignment played an essential role in regulating FN alignment in response to shear stress. Our findings offer insight into how ECs cooperatively remodel their subcellular ECM and intracellular F-actin cytoskeleton in response to mechanical stimuli, and are valuable for vascular tissue engineering.

Type
Biological Science Applications
Copyright
© Microscopy Society of America 2017 

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Footnotes

a

Xianghui Gong and Xixi Zhao contributed equally to this work.

References

Babatunde, I.G., Charles, A.O., Kayode, A.B. & Olatubosun, O. (2012). Fingerprint image enhancement: Segmentation to thinning. Int J Adv Comput Sci Appl 3(1), 322325.Google Scholar
Balcioglu, H.E., van Hoorn, H., Donato, D.M., Schmidt, T. & Danen, E.H. (2015). The integrin expression profile modulates orientation and dynamics of force transmission at cell-matrix adhesions. J Cell Sci 128(7), 13161326.CrossRefGoogle ScholarPubMed
Bourget, J.-M., Guillemette, M., Veres, T., Auger, F.A. & Germain, L. (2013). Alignment of Cells and Extracellular Matrix Within Tissue-Engineered Substitutes. Rijeka: INTECH Open Access Publisher.CrossRefGoogle Scholar
Bray, M.A., Adams, W.J., Geisse, N.A., Feinberg, A.W., Sheehy, S.P. & Parker, K.K. (2010). Nuclear morphology and deformation in engineered cardiac myocytes and tissues. Biomaterials 31(19), 51435150.CrossRefGoogle ScholarPubMed
Burridge, K., Fath, K., Kelly, T., Nuckolls, G. & Turner, C. (1988). Focal adhesions: Transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Dev Biol 4(4), 487525.CrossRefGoogle ScholarPubMed
Byung-Gyu, K., Han-Ju, K. & Dong-Jo, P. (2002). New enhancement algorithm for fingerprint images. Proc IEEE 4651(2), 10511055.Google Scholar
Canver, A.C. & Morss Clyne, A. (2017). Quantification of multicellular organization, junction integrity, and substrate features in collective cell migration. Microsc Microanal 23(1), 2233.CrossRefGoogle ScholarPubMed
Chen, Z., Givens, C., Reader, J.S. & Tzima, E. (2017). Haemodynamics regulate fibronectin assembly via PECAM. Sci Rep 7, 41223.CrossRefGoogle ScholarPubMed
Dartsch, P.C. & Betz, E. (1989). Response of cultured endothelial cells to mechanical stimulation. Basic Res Cardiol 84(3), 268281.CrossRefGoogle ScholarPubMed
Figueroa, D.S., Kemeny, S.F. & Clyne, A.M. (2014). Glycated collagen decreased endothelial cell fibronectin alignment in response to cyclic stretch via interruption of actin alignment. J Biomech Eng 136(10), 101010.CrossRefGoogle ScholarPubMed
Fletcher, D.A. & Mullins, R.D. (2010). Cell mechanics and the cytoskeleton. Nature 463(7280), 485492.CrossRefGoogle ScholarPubMed
Fuseler, J.W., Millette, C.F., Davis, J.M. & Carver, W. (2007). Fractal and image analysis of morphological changes in the actin cytoskeleton of neonatal cardiac fibroblasts in response to mechanical stretch. Microsc Microanal 13(2), 133143.CrossRefGoogle ScholarPubMed
Gardel, M.L., Schneider, I.C., Aratyn-Schaus, Y. & Waterman, C.M. (2010). Mechanical integration of actin and adhesion dynamics in cell migration. Annu Rev Cell Dev Biol 26(1), 315333.CrossRefGoogle ScholarPubMed
Geiger, B., Bershadsky, A., Pankov, R. & Yamada, K.M. (2001). Transmembrane crosstalk between the extracellular matrix – Cytoskeleton crosstalk. Nat Rev Mol Cell Biol 2(11), 793805.CrossRefGoogle ScholarPubMed
Girard, P.R. & Nerem, R.M. (1995). Shear stress modulates endothelial cell morphology and F-actin organization through the regulation of focal adhesion-associated proteins. J Cell Physiol 163(1), 179193.CrossRefGoogle ScholarPubMed
Gong, X., Liu, H., Ding, X., Liu, M., Li, X., Zheng, L., Jia, X., Zhou, G., Zou, Y., Li, J., Huang, X. & Fan, Y. (2014). Physiological pulsatile flow culture conditions to generate functional endothelium on a sulfated silk fibroin nanofibrous scaffold. Biomaterials 35(17), 47824791.CrossRefGoogle ScholarPubMed
Gong, X., Yao, J., He, H., Zhao, X., Liu, X., Zhao, F., Sun, Y. & Fan, Y. (2017). Combination of flow and micropattern alignment affecting flow-resistant endothelial cell adhesion. J Mech Behav Biomed Mater 74, 1120.CrossRefGoogle ScholarPubMed
Humphries, M.J., Travis, M.A., Clark, K. & Mould, A.P. (2004). Mechanisms of integration of cells and extracellular matrices by integrins. Biochem Soc Trans 32(5), 822825.CrossRefGoogle ScholarPubMed
Hynes, R.O. & Destree, A.T. (1978). Relationships between fibronectin (LETS protein) and actin. Cell 15(3), 875886.CrossRefGoogle ScholarPubMed
Jinguji, Y. & Fujiwara, K. (1994). Stress fiber dependent axial organization of fibronectin fibrils in the basal lamina of the chick aorta and mesenteric artery. Endothelium 2(1), 3547.CrossRefGoogle Scholar
Kemeny, S.F. & Clyne, A.M. (2011). A simplified implementation of edge detection in MATLAB is faster and more sensitive than Fast Fourier Transform for Actin fiber alignment quantification. Microsc Microanal 17(2), 156166.CrossRefGoogle ScholarPubMed
Lemmon, C.A., Chen, C.S. & Romer, L.H. (2009). Cell traction forces direct fibronectin matrix assembly. Biophys J 96(2), 729738.CrossRefGoogle ScholarPubMed
Lin, H., Yifei, W. & Jain, A. (1998). Fingerprint image enhancement: Algorithm and performance evaluation. IEEE Trans Pattern Anal Mach Intell 20(8), 777789.Google Scholar
Malek, A.M. & Izumo, S. (1996). Mechanism of endothelial cell shape change and cytoskeletal remodeling in response to fluid shear stress. J Cell Sci 109(4), 713726.CrossRefGoogle ScholarPubMed
Marquez, J.P. (2006). Fourier analysis and automated measurement of cell and fiber angular orientation distributions. Int J Solids Struct 43(21), 64136423.CrossRefGoogle Scholar
Marr, D. & Hildreth, E. (1980). Theory of edge detection. Proc R Soc London 207(1167), 187217.Google ScholarPubMed
McCue, S., Noria, S. & Langille, B.L. (2004). Shear-induced reorganization of endothelial cell cytoskeleton and adhesion complexes. Trends Cardiovas Med 14(4), 143151.CrossRefGoogle ScholarPubMed
Ng, C.P., Hinz, B. & Swartz, M.A. (2005). Interstitial fluid flow induces myofibroblast differentiation and collagen alignment in vitro. J Cell Sci 118(20), 47314739.CrossRefGoogle ScholarPubMed
Palmer, B.M. & Bizios, R. (1997). Quantitative characterization of vascular endothelial cell morphology and orientation using Fourier transform analysis. J Biomech Eng 119(2), 159165.CrossRefGoogle ScholarPubMed
Pong, T., Adams, W.J., Bray, M.A., Feinberg, A.W., Sheehy, S.P., Werdich, A.A. & Parker, K.K. (2011). Hierarchical architecture influences calcium dynamics in engineered cardiac muscle. Exp Biol Med 236(3), 366373.CrossRefGoogle ScholarPubMed
Rao, A.R. (1990). A taxonomy for texture description and identification. Berlin: Springer.CrossRefGoogle Scholar
Resnick, N., Yahav, H., Shay-Salit, A., Shushy, M., Schubert, S., Zilberman, L.C.M. & Wofovitz, E. (2003). Fluid shear stress and the vascular endothelium: For better and for worse. Prog Biophys Mol Biol 81(3), 177199.CrossRefGoogle ScholarPubMed
Rozario, T., Dzamba, B., Weber, G.F., Davidson, L.A. & Desimone, D.W. (2009). The physical state of fibronectin matrix differentially regulates morphogenetic movements in vivo. Dev Biol 327(2), 386398.CrossRefGoogle ScholarPubMed
Smith, J.T.G., Marks, W.B., Lange, G.D. Jr, Sheriff, W. & Neale, E.A. (1988). Edge detection in images using Marr-Hildreth filtering techniques. J Neurosci Methods 26(1), 7581.CrossRefGoogle ScholarPubMed
Sugimoto, K., Fujii, S., Takemasa, T. & Yamashita, K. (1997). Factors inducing codistribution of marginal actin fibers and fibronectin in rat aortic endothelial cells. Am J Physiol 272(2), H2188H2194.Google ScholarPubMed
Thoumine, O., Nerem, R.M. & Girard, P.R. (1995). Changes in organization and composition of the extracellular matrix underlying cultured endothelial cells exposed to laminar steady shear stress. Lab Invest 73(4), 565576.Google ScholarPubMed
Tzima, E., del Pozo, M.A., Shattil, S.J., Chien, S. & Schwartz, M.A. (2001). Activation of integrins in endothelial cells by fluid shear stress mediates Rho-dependent cytoskeletal alignment. EMBO J 20(17), 46394647.CrossRefGoogle ScholarPubMed
Vartanian, K.B., Kirkpatrick, S.J., Hanson, S.R. & Hinds, M.T. (2008). Endothelial cell cytoskeletal alignment independent of fluid shear stress on micropatterned surfaces. Biochem Biophys Res Commun 371(4), 787792.CrossRefGoogle ScholarPubMed
Wechezak, A., Viggers, R. & Sauvage, L. (1985). Fibronectin and F-actin redistribution in cultured endothelial cells exposed to shear stress. Lab Invest 53(6), 639647.Google ScholarPubMed
Yoshigi, M., Clark, E.B. & Yost, H.J. (2003). Quantification of stretch-induced cytoskeletal remodeling in vascular endothelial cells by image processing. Cytometry A 55(2), 109118.CrossRefGoogle ScholarPubMed
Zhong, C., Chrzanowskawodnicka, M., Brown, J., Shaub, A., Belkin, A.M. & Burridge, K. (1998). Rho-mediated contractility exposes a cryptic site in fibronectin and induces fibronectin matrix assembly. J Cell Biol 141(2), 539551.CrossRefGoogle ScholarPubMed
Zhou, X., Rowe, R.G., Hiraoka, N., George, J.P., Wirtz, D., Mosher, D.F., Virtanen, I., Chernousov, M.A. & Weiss, S.J. (2008). Fibronectin fibrillogenesis regulates three-dimensional neovessel formation. Genes Dev 22(9), 12311243.CrossRefGoogle ScholarPubMed