Cell prestress. II. Contribution of microtubules.
The tensegrity model hypothesizes that cytoskeleton-based microtubules (MTs) carry compression as they balance a portion of cell contractile stress. To test this hypothesis, we used traction force microscopy to measure traction at the interface of adhering human airway smooth muscle cells and a flexible polyacrylamide gel substrate. The prediction is that if MTs balance a portion of contractile stress, then, upon their disruption, the portion of stress balanced by MTs would shift to the substrate, thereby causing an increase in traction. Measurements were done first in maximally activated cells (10 microM histamine) and then again after MTs had been disrupted (1 microM colchicine). We found that after disruption of MTs, traction increased on average by approximately 13%. Because in activated cells colchicine induced neither an increase in intracellular Ca(2+) nor an increase in myosin light chain phosphorylation as shown previously, we concluded that the observed increase in traction was a result of load shift from MTs to the substrate. In addition, energy stored in the flexible substrate was calculated as work done by traction on the deformation of the substrate. This result was then utilized in an energetic analysis. We assumed that cytoskeleton-based MTs are slender elastic rods supported laterally by intermediate filaments and that MTs buckle as the cell contracts. Using the post-buckling equilibrium theory of Euler struts, we found that energy stored during buckling of MTs was quantitatively consistent with the measured increase in substrate energy after disruption of MTs. This is further evidence supporting the idea that MTs are intracellular compression-bearing elements.
Stamenović
Dimitrije
D
Department of Biomedical Engineering, Boston University, 44 Cummington Street, Boston, MA 02215, USA. dimitrij@engc.bu.edu
Mijailovich
Srboljub M
SM
Tolić-Nørrelykke
Iva Marija
IM
Chen
Jianxin
J
Wang
Ning
N
D000180:Acrylic Resins / Q000737:chemistry; D002448:Cell Adhesion / Q000502:physiology*; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods; D008870:Microtubules / Q000502:physiology*; D008954:Models, Biological; D009119:Muscle Contraction / Q000502:physiology; D009130:Muscle, Smooth / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D013314:Stress, Mechanical; D013718:Tensile Strength
D000180:Acrylic Resins; C016680:polyacrylamide gels; D003078:Colchicine