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Fractal and Image Analysis of Morphological Changes in the Actin Cytoskeleton of Neonatal Cardiac Fibroblasts in Response to Mechanical Stretch

Published online by Cambridge University Press:  19 March 2007

John W. Fuseler ,
Clarke F. Millette ,
Jeffery M. Davis  and
Wayne Carver
John W. Fuseler
Affiliation:
Department of Cell and Developmental Biology and Anatomy, University of South Carolina, School of Medicine, Columbia, South Carolina 29209, USA
Clarke F. Millette
Affiliation:
Department of Cell and Developmental Biology and Anatomy, University of South Carolina, School of Medicine, Columbia, South Carolina 29209, USA
Jeffery M. Davis
Affiliation:
Department of Cell and Developmental Biology and Anatomy, University of South Carolina, School of Medicine, Columbia, South Carolina 29209, USA
Wayne Carver
Affiliation:
Department of Cell and Developmental Biology and Anatomy, University of South Carolina, School of Medicine, Columbia, South Carolina 29209, USA
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Abstract

Cardiac fibroblasts are the most numerous cells in the heart and are critical in the formation and normal functioning of the organ. Cardiac fibroblasts are firmly attached to and surrounded by extracellular matrix (ECM). Mechanical forces transmitted through interaction with the ECM can result in changes of overall cellular shape, cytoskeletal organization, proliferation, and gene expression of cardiac fibroblasts. These responses may be different in the normally functioning heart, when compared with various pathological conditions, including inflammation or hypertrophy. It is apparent that cellular phenotype and physiology, in turn, are affected by multiple signal transduction pathways modulated directly by the state of polymerization of the actin cytoskeleton. Morphological changes in actin organization resulting from response to adverse conditions in fibroblasts and other cell types are basically descriptive. Some studies have approached quantifying changes in actin cytoskeletal morphology, but these have involved complex and difficult procedures. In this study, we apply image analysis and non-Euclidian geometrical fractal analysis to quantify and describe changes induced in the actin cytoskeleton of cardiac fibroblasts responding to mechanical stress. Characterization of these rapid responses of fibroblasts to mechanical stress may provide insight into the regulation of fibroblasts behavior and gene expression during heart development and disease.

Keywords

cardiac fibroblastsactin cytoskeletonfractal geometryimage analysisimmunofluorescencefluorescence microscopymechanical stretch
Type
BIOLOGICAL APPLICATIONS
Information
Microscopy and Microanalysis , Volume 13 , Issue 2 , April 2007 , pp. 133 - 143
Copyright
© 2007 Microscopy Society of America

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References

REFERENCES

Anderson, J.C., Babb, A.L. & Hlastala, M.P. (2005). A fractal analysis of the radial distribution of bronchial capillaries around large airways. J Appl Physiol 98, 850–855.Google Scholar
Baish, J.W. & Jain, R.K. (2000). Fractals and cancer. Cancer Res 60, 3683–3688.Google Scholar
Behar, T.N. (2001). Analysis of fractal dimension of O2A glial cells differentiating in vitro. Methods 24, 331–339.Google Scholar
Bernard, F., Bossu, J.L. & Gaillard, S. (2001). Identification of living oligodendrocyte developmental stages by fractal analysis of cell morphology. J Neurosci Res 65, 439–445.Google Scholar
Bishop, J.E. (1998). Regulation of cardiovascular collagen deposition by mechanical forces. Mol Med Today 4, 69–75.Google Scholar
Bona, A.D., Hill, T.J. & Mecholsky, J.J., Jr. (2001). The effect of contour angle on fractal dimension measurements for brittle materials. J Material Sci 36, 2645–2650.Google Scholar
Borg, T.K. & Caulfield, J.B. (1981). The collagen matrix of the heart. Fed Proc 40, 2037–2041.Google Scholar
Borg, T.K., Rubin, K., Lundgren, E., Borg, K. & Obrink, B. (1984). Recognition of extracellular matrix components by neonatal and adult cardiac myocytes. Dev Biol 104, 86–96.Google Scholar
Borodinsky, L.N. & Fiszman, M.L. (2001). A single-cell model to study changes in neuronal fractal dimension. Methods 24, 341–345.Google Scholar
Caldwell, C.B., Moran, E.L. & Bogoch, E.R. (1998). Fractal dimension as a measure of altered trabecular bone in experimental inflammatory arthritis. J Bone Miner Res 13, 978–985.Google Scholar
Camelliti, P., Borg, T.K. & Kohl, P. (2005). Structural and functional characterization of cardiac fibroblasts. Cardiovasc Res 65, 40–51.Google Scholar
Carver, W., Nagpal, M.L., Nachtigal, M., Borg, T.K. & Terracio, L. (1991). Collagen expression in mechanically stimulated cardiac fibroblasts. Circ Res 69, 116–122.Google Scholar
Carver, W., Terracio, L. & Borg, T.K. (1993). Expression and accumulation of interstitial collagen in the neonatal rat heart. Anat Rec 236, 511–520.Google Scholar
Caserta, F., Eldred, W.D., Fernandez, E., Hausman, R.E., Stanford, L.R., Bulderev, S.V., Schwarzer, S. & Stanley, H.E. (1995). Determination of fractal dimension of physiologically characterized neurons in two and three dimensions. J Neurosci Methods 56, 133–144.Google Scholar
Costa, K.D., May-Newman, K., Farr, D. & O'Dell, W.G. (1997). Three dimensional residual strain in midanterior canine left ventricle. Am J Physiol 273, H1968–H1976.Google Scholar
Cross, S.S. (1997). Fractals in pathology. J Pathol 182, 1–8.Google Scholar
DeMeester, S.L., Cobb, J.P., Hotchkiss, R.S., Osborne, D.F., Karl, I.E., Tinsley, K.W. & Buchman, T.G. (1998). Stress-induced fractal rearrangement of the endothelial cell cytoskeleton causes apoptosis. Surgery 124, 362–371.Google Scholar
Fernandez, E. & Jelinek, H.F. (2001). Use of fractal theory in neuroscience: Methods, advantages, and potential problems. Methods 24, 309–321.Google Scholar
Fringer, J. & Grinnell, F. (2003). Fibroblast quiescence in floating collagen matrices. J Biol Chem 78, 20612–206117.Google Scholar
Gazit, Y., Baish, J.W., Safabakhsh, N., Leunig, M., Baxter, L.T. & Jain, R.K. (1997). Fractal characteristics of tumor vascular architecture during tumor growth and regression. Microcirculation 4, 395–402.Google Scholar
Gazit, Y., Berk, D.A., Leunig, M., Baxter, L.T. & Jain, R.K. (1995). Scale-invariant behavior and vascular network formation in normal and tumor tissue. Phys Rev Lett 75, 2428–2431.Google Scholar
Geiger, B. & Bershadsky, A. (2001). Assembly and mechanosensory function of focal contacts. Curr Opin Cell Biol 13, 584–592.Google Scholar
Glenny, R.W., Robertson, H.T., Yamashiro, S. & Bassingthwaighte, J.B. (1991). Applications of fractal analysis to physiology. J Appl Physiol 70, 2351–2367.Google Scholar
Goldsmith, E.C., Hoffman, A., Morales, M.O., Pott, J.D., Price, R.L., McFadden, A., Rice, M. & Borg, T.K. (2004). Organization of fibroblasts in the heart. Develop Dyn 230, 787–794.Google Scholar
Grinnell, F. (2000). Fibroblast-collagen matrix contraction: Growth factor signaling and mechanical loading. Trends Cell Biol 10, 362–365.Google Scholar
Grinnell, F. (2003). Fibroblast biology in three-dimensional collagen matrices. Trends Cell Biol 13, 264–269.Google Scholar
Grinnell, F. & Ho, C.-H. (2002). Transforming growth factor beta stimulates fibroblast-collagen matrix contraction by different mechanisms in mechanically loaded and unloaded matrices. Expt Cell Res 273, 248–255.Google Scholar
Grizzi, F., Russo, C., Colombo, P., Franceschini, B., Frezza, E.E., Cobos, E. & Chiriva-Internati, M. (2005). Quantitative evaluation and modeling of two-dimensional neovascular network complexity: The surface fractal dimension. BMC Cancer 5, 14–23.Google Scholar
Hanaichi, T., Sato, T., Iwamoto, T., Malavasi-Yamashiro, J., Hoshino, M. & Mizuno, N. (1986). A stable lead by modification of Sato's method. J Electron Microsc (Tokyo) 35, 304–306.Google Scholar
Landini, G. (1996). Applications of fractal geometry in pathology. In Fractal Geometry in Biological Systems, An Analytical Approach, Iannaccone, P.M. & Khokha, M. (Eds.), pp. 205–246. New York: CRC Press.
Lee, A.A., Delhaas, T., McCulloch, A.D. & Villarreal, F.J. (1999). Differential responses of adult cardiac fibroblasts to in vitro biaxial strain patterns. J Mol Cell Cardiol 31, 1833–1843.Google Scholar
Lee, A.A., Delhaas, T., Waldman, L.K., MacKenna, D.A., Villarreal, F.J. & McCulloch, A.D. (1996). An equibiaxial strain system for cultured cells. Am J Physiol 271, C1400–C1408.Google Scholar
Lovejoy, S. (1982). Area-perimeter relation for rain and cloud areas. Science 216, 185–187.Google Scholar
MacKenna, D., Summerour, S.R. & Villarreal, F.J. (2000). Role of mechanical factors in modulating cardiac fibroblast function and extracellular matrix synthesis. Cardiovasc Res 46, 257–263.Google Scholar
Mandelbrot, B.B., Passoja, D.E. & Paully, A.J. (1984). Fractal character of fractures surfaces of metals. Nature 308, 721–724.Google Scholar
Milosevic, N.T., Ristanovic, D. & Stankovic, J.B. (2005). Fractal analysis of the laminar organization of spinal cord neurons. J Neurosci Methods 146, 198–204.Google Scholar
Molina, T., Kabsch, K., Alonso, A., Kohl, A., Kompossch, G. & Tomakidi, P. (2001). Topographic changes of focal adhesion components and modulation of p125FAK activation in stretched human periodontal ligament fibroblasts. J Dent Res 80, 1984–1989.Google Scholar
Montague, P.R. & Friedlander, M.J. (1991). Morphogenesis and territorial coverage by isolated mammalian retinal ganglion cells. J Neurosci 11, 1440–1457.Google Scholar
Munevar, S., Wang, Y.L. & Dembo, M. (2004). Regulation of mechanical interactions between fibroblasts and the substratum by stretch-activated Ca2+ entry. J Cell Sci 117, 85–92.Google Scholar
Nezadal, M., Zemeskal, O. & Buchnicek, M. (2001). The box-counting: critical study. In 4th Conference on Prediction, Synergetic and more. The Faculty of Management, Institute of Information Technologies, Faculty of Technology, Tomas Bata University in Zlin, October 25–26. p. 18. HarFA software (http://www.fch.vutbr.cz/lectures/imagesci).
Ramon-Moliner, E. (1962). An attempt at classifying nerve cells on the basis of their dendritic patterns. J Comp Neurol 119, 211–227.Google Scholar
Rexed, B. (1952). The cytoarchitectonic organization of the spinal cord in the cat. J Comp Neurol 96, 415–496.Google Scholar
Rexed, B. (1954). The cytoarchitectonic atlas of the spinal cord in the cat. J Comp Neurol 100, 297–380.Google Scholar
Ristanovic, D., Nedeljkov, V., Stefanovic, B.D., Milosevic, N.T., Grgurevic, M. & Stulic, V. (2002). Fractal and nonfractal analysis of cell images: Comparison and application to neuronal dendritic arborization. Biol Cybern 87, 278–288.Google Scholar
Ruwhof, C. & van der Laarse, A. (2000). Mechanical stress-induced cardiac hypertrophy: Mechanisms and signal transduction pathways. Cardiovas Res 47, 23–37.Google Scholar
Sedivy, R., Thurner, S., Budinsky, A.C., Kostler, W.J. & Zielinski, C.C. (2002). Short-term rhythmic proliferation of human breast cancer cell lines: Surface effects and fractal growth patterns. J Pathol 197, 163–169.Google Scholar
Sorescu, D. & Griendling, K.K. (2002). Reactive oxygen species, mitochondria, and NAD(P)H oxidases in the development and progression of heart failure. Congest Heart Failure 8, 132–140.Google Scholar
Sussman, M.A., McCulloch, A. & Borg, T.K. (2002). Dance band on the Titanic. Biomechanical signaling in cardiac hypertrophy. Circ Res 91, 888–898.Google Scholar
Tamariz, E. & Grinnell, F. (2002). Modulation of fibroblast morphology and adhesion during collagen matrix remodeling. Mol Biol Cell 13, 3915–3929.Google Scholar
Thomason, D.B., Anderson, O., 3rd & Menon, V. (1996). Fractal analysis of cytoskeleton rearrangement in cardiac muscle during head-down tilt. J Appl Physiol 81, 1522–1527.Google Scholar
Tian, B., Liu, J., Bitterman, P. & Bache, R.J. (2003). Angiotensin II modulates nitric oxide-induced cardiac fibroblast apoptosis by activation of AKT/PKB. Am J Physiol Heart Circ Physiol 285, H1105–H1112.Google Scholar
Vasiliev, J.M. (1991). Polarization of pseudopodial activities: Cytoskeletal mechanisms. J Cell Sci 98, 1–4.Google Scholar
Walter, R.J., Jr. & Berns, M.W. (1986). Digital image processing and analysis. In Video Microscopy, Inoue, S. (Ed.), pp 327–392. New York and London: Plenum Press.
Weber, K.T. (1989). Cardiac interstitium in health and disease: The fibrillar collagen network. J Am Coll Cardiol 13, 1637–1652.Google Scholar
Wick, N., Thurner, S., Paiha, K., Sedivy, R., Vietor, I. & Huber, L.A. (2003). Quantitative measurement of cell migration using time-lapse videomicroscopy and non-linear system analysis. Histochem Cell Biol 119, 15–20.Google Scholar
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, 109–118.Google Scholar
Zhang, L., Liu, J.Z., Dean, D., Sahgal, V. & Yue, G.H. (2005). A three dimensional fractal analysis method for quantifying white matter structure in human brain. J Neurosci Methods 150, 242–253.Google Scholar