Direct measures of large, anisotropic strains in deformation of the erythrocyte cytoskeleton.
The erythrocyte's spectrin-actin membrane skeleton is directly shown to be capable of sustaining large, anisotropic strains. Photobleaching of an approximately 1-micrometer stripe in rhodamine phalloidin-labeled actin appears stable up to at least 37 degrees C, and is used to demonstrate large in-surface stretching during elastic deformation of the skeleton. Principal extension or stretch ratios of at least approximately 200% and contractions down to approximately 40%, both referenced to an essentially undistorted cell, are visually demonstrated in micropipette-imposed deformation. Such anisotropic straining is seen to be consistent at a qualitative level with now classic analyses (Evans. 1973. Biophys. J. 13:941-954) and is generally nonhomogeneous though axisymmetric down to the submicron scale. Local, direct measurements of stretching prove quantitatively consistent (within approximately 10%) with integrated estimates that are based simply on a measured relative density distribution of actin. The measurements are also in close agreement with direct computation of mean spectrin chain extension in full statistical mechanical simulations of a coarse-grained network held in a micropipette. Finally, as a cell thermally fragments near approximately 48 degrees C, the patterned photobleaching demonstrates a destructuring of the surface network in a process that is more readily attributable to transitions in spectrin than in F-actin.
Lee
J C
JC
Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Wong
D T
DT
Discher
D E
DE
D000199:Actins / Q000737:chemistry; D055592:Biophysical Phenomena; D001703:Biophysics; D003599:Cytoskeleton / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D004907:Erythrocyte Deformability / Q000502:physiology*; D004912:Erythrocytes / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D006801:Humans; D066298:In Vitro Techniques; D008954:Models, Biological; D010590:Phalloidine; D012235:Rhodamines; D013049:Spectrin / Q000737:chemistry; D013314:Stress, Mechanical; D013696:Temperature
D000199:Actins; D005456:Fluorescent Dyes; D012235:Rhodamines; D013049:Spectrin; D010590:Phalloidine