Muscle cell peeling from micropatterned collagen: direct probing of focal and molecular properties of matrix adhesion.
To quantitatively elucidate attributes of myocyte-matrix adhesion, muscle cells were controllably peeled from narrow strips of collagen-coated glass. Initial growth of primary quail myoblasts on collagen strips was followed by cell alignment, elongation and end-on fusion between neighbors. This geometric influence on differentiation minimized lateral cell contact and cell branching, enabling detailed study of myocyte-matrix adhesion. A micropipette was used to pull back one end of a quasi-cylindrical cell while observing in detail the non-equilibrium detachment process. Peeling velocities fluctuated as focal roughness, microm in scale, was encountered along the detachment front. Nonetheless, mean peeling velocity ( microm/second) generally increased with detachment force (nN), consistent with forced disruption of adhesion bonds. Immunofluorescence of beta1-integrins correlated with the focal roughness and appeared to be clustered in axially extended focal contacts. In addition, the peeling forces and rates were found to be moderately well described by a dynamical peeling model for receptor-based adhesion (Dembo, M., Torney, D. C., Saxman, K. and Hammer, D. (1988). Proc. R. Soc. Lond. B 234, 55-83). Estimates were thereby obtained for the spontaneous, molecular off-rate (kooff, (less than or equal to)10/seconds) and the receptor complex stiffness (kappa, approx. 10(-5)-10(-6) N/m) of adherent myocytes. Interestingly, the local stiffness is within the range of flexible proteins of the spectrin superfamily. The overall approach lends itself to elucidating the developing function of other structural and adhesive components of cells, particularly skeletal muscle cells with specialized components, such as the spectrin-homolog dystrophin and its membrane-linked receptor dystroglycan.
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Biotechnology Program, School of Engineering and Applied Science, Dept of Physiology, Pennsylvania Muscle Institute and Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, PA 19104, USA.
Picart
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Feng
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Sweeney
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Discher
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D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005898:Glass; D008954:Models, Biological; D009132:Muscles / Q000378:metabolism*; D051381:Rats; D013997:Time Factors
D003094:Collagen