Mechano-chemical control of human endothelium orientation and size

Human umbilical vein endothelial cells (EC) were grown on elastic silicone membranes subjected to cyclic stretch, simulating arterial wall motion. Stretching conditions (20% amplitude, 52 cycle/min) stimulated stress fiber formation and their orientation transversely to the strain direction. Cell bodies aligned along the same axis after the actin cytoskeleton. EC orientation response was inhibited by the adenylate cyclase activator, forskolin (10(-5) M), which caused stress fiber disassembly and the redistribution of F-actin to the cortical cytoplasm. Preoriented EC depleted of stress fibers by forskolin treatment retained their aligned state. Thus, stress fibers are essential for the process of EC orientation induced by repeated strain, but not for the maintenance of EC orientation. The monolayer formed by EC grown to confluence in conditions of intermittent strain consisted of uniform elongated cells and was resistant to deformation. In contrast, the monolayer assembled in stationary conditions was less compliant and exposed local denudations on initiation of stretching. When stretched in the presence of 10(-5) M forskolin it rapidly (3-4 h) reestablished integrity but gained a heterogeneous appearance since denuded areas were covered by giant cells. The protective effect of forskolin was because of the stimulation of EC spreading. This feature of forskolin was demonstrated while studying its action on EC spreading and repair of a scratched EC monolayer in conventional culture. Thus mechanical deformation and adenylate cyclase activity may be important factors in the control of endothelium morphology in human arteries.

[1]  C. Klee,et al.  Phosphorylation of smooth muscle myosin light chain kinase by the catalytic subunit of adenosine 3': 5'-monophosphate-dependent protein kinase. , 1978, The Journal of biological chemistry.

[2]  T. Pollard,et al.  Actin filament stress fibers in vascular endothelial cells in vivo. , 1983, Science.

[3]  F. Booyse,et al.  Culture of Arterial Endothelial Cells , 1975, Thrombosis and Haemostasis.

[4]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[5]  V. Caviness,et al.  HEMORRHAGES INTO PERIPHERAL NERVES IN ASSOCIATION WITH LEUKEMIA. , 1964 .

[6]  R. Franke,et al.  Induction of human vascular endothelial stress fibres by fluid shear stress , 1984, Nature.

[7]  J. Scholey,et al.  Regulation of non-muscle myosin assembly by calmodulin-dependent light chain kinase , 1980, Nature.

[8]  Taylor Rg,et al.  Endothelial surface characteristics in pigeon coronary artery atherosclerosis. I. Cellular alterations during the initial stages of dietary cholesterol challenge. , 1982 .

[9]  R. Taylor,et al.  Endothelial surface characteristics in pigeon coronary artery atherosclerosis. I. Cellular alterations during the initial stages of dietary cholesterol challenge. , 1982, Laboratory investigation; a journal of technical methods and pathology.

[10]  K. Weber,et al.  Calcium control of actin-myosin based contraction in triton models of mouse 3T3 fibroblasts is mediated by the myosin light chain kinase (MLCK)-calmodulin complex. , 1983, Experimental cell research.

[11]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[12]  Chazov Ei,et al.  Vessel Wall in Athero- and Thrombogenesis , 1982, Springer Berlin Heidelberg.

[13]  A. M. Vikhert,et al.  On the Endothelial Lining of Human Arteries at Genesis of the Atherosclerotic Plaque , 1982 .

[14]  S. Singer,et al.  Intracellular distributions of mechanochemical proteins in cultured fibroblasts. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[15]  G. Osikowicz,et al.  Effects of chronic oral consumption of nicotine on the rabbit aortic endothelium. , 1981, The American journal of pathology.

[16]  V. Babaev,et al.  Primary culture of endothelial cells from atherosclerotic human aorta. Part 1. Identification, morphological and ultrastructural characteristics of two endothelial cell subpopulations. , 1986, Atherosclerosis.

[17]  R. Buck,et al.  Reorientation response of cells to repeated stretch and recoil of the substratum. , 1980, Experimental cell research.

[18]  R. Buck.  Behavior of vascular smooth muscle cells during repeated stretching of the substratum in vitro. , 1983, Atherosclerosis.

[19]  H. Hämmerle,et al.  Orientation of cultured arterial smooth muscle cells growing on cyclically stretched substrates. , 1986, Acta anatomica.

[20]  Vladimir Gelfand,et al.  Organization of stress fibers in cultured fibroblasts after extraction of actin with bovine brain gelsolin-like protein. , 1987, Experimental cell research.

[21]  V. Tkachuk,et al.  Morphological alterations in endothelial cells from human aorta and umbilical vein induced by forskolin and phorbol 12-myristate 13-acetate: a synergistic action of adenylate cyclase and protein kinase C activators. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Gotlieb,et al.  Endothelial Cell Monolayer Integrity: I. Characterization of Dense Peripheral Band of Microfilaments , 1986, Arteriosclerosis.

[23]  R L Kormos,et al.  Hemodynamics and the vascular endothelial cytoskeleton , 1987, The Journal of cell biology.

[24]  A. Antonov,et al.  Heterogeneity of endothelium in human aorta. A quantitative analysis by scanning electron microscopy. , 1984, Atherosclerosis.

[25]  H. Hämmerle,et al.  Orientation response of arterial smooth muscle cells to mechanical stimulation. , 1986, European journal of cell biology.

[26]  C F Dewey,et al.  The dynamic response of vascular endothelial cells to fluid shear stress. , 1981, Journal of biomechanical engineering.