Acto-myosin cytoskeleton dependent viscosity and shear-thinning behavior of the amoeba cytoplasm.
The mechanical behavior of the human parasite Entamoeba histolytica plays a major role in the invasive process of host tissues and vessels. In this study, we set up an intracellular rheological technique derived from magnetic tweezers to measure the viscoelastic properties within living amoebae. The experimental setup combines two magnetic fields at 90 degrees from each other and is adapted to an inverted microscope, which allows monitoring of the rotation of pairs of magnetic phagosomes. We observe either the response of the phagosome pair to an instantaneous 45 degrees rotation of the magnetic field or the response to a permanent uniform rotation of the field at a given frequency. By the first method, we concluded that the phagosome pairs experience a soft viscoelastic medium, represented by the same mechanical model previously described for the cytoplasm of Dictyostelium discoideum [Feneberg et al. in Eur Biophys J 30(4):284-294 2001]. By the second method, the permanent rotation of a pair allowed us to apply a constant shear rate and to calculate the apparent viscosity of the cytoplasm. As found for entangled polymers, the viscosity decreases with the shear rate applied (shear-thinning behavior) and exhibits a power-law-type thinning, with a corresponding exponent of 0.65. Treatment of amoeba with drugs that affect the actin polymer content demonstrated that the shear-thinning behavior of the cytoplasm depends on the presence of an intact actin cytoskeleton. These data present a physiologic relevance for Entamoeba histolytica virulence. The shear-thinning behavior could facilitate cytoplasm streamings during cell movement and cell deformation, under important shear experienced by the amoeba during the invasion of human tissues. In this study, we also investigated the role of the actin-based motor myosin II and concluded that myosin II stiffens the F-actin gel in living parasites likely by its cross-linking activity.
Marion
Sabrina
S
Unité de Biologie Cellulaire du Parasitisme, INSERM U389, Institut Pasteur, Paris, France.
Guillen
Nancy
N
Bacri
Jean-Claude
JC
Wilhelm
Claire
C
D000199:Actins / Q000502:physiology*; D000205:Actomyosin / Q000502:physiology*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000502:physiology*; D004748:Entamoeba histolytica / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods; D008280:Magnetics; D008846:Micromanipulation / Q000379:methods*; D020409:Molecular Motor Proteins / Q000502:physiology*; D024462:Myosin Type II / Q000502:physiology*; D010812:Physical Stimulation / Q000379:methods; D033081:Shear Strength; D013314:Stress, Mechanical; D014783:Viscosity
D000199:Actins; D020409:Molecular Motor Proteins; D000205:Actomyosin; D024462:Myosin Type II