The biased lamellipodium development and microtubule organizing center position in vascular endothelial cells migrating under the influence of fluid flow.
To analytically study the morphological responses of vascular endothelial cells (ECs) to fluid flow, we designed a parallel plate flow culture chamber in which cells were cultured under fluid shear stress ranging from 0.01 to 2.0 Pa for several days. Via a viewing window of the chamber, EC responses to known levels of fluid shear stress were monitored either by direct observations or by a video-enhanced time-lapse microscopy. Among the responses of cultured ECs to flow, morphological responses take from hours to days to be fully expressed, except for the fluid shear stress-dependent motility pattern change we reported earlier which could be detected within 30 min of flow changes. We report here that ECs exposed to more than 1.0 Pa of fluid shear stress have developed lamellipodia in the direction of flow in 10 min. This is the fastest structurally identifiable EC response to fluid shear stress. This was a reversible response. When the flow was stopped or reduced to the level which exerted less than 0.1 Pa of fluid shear stress, the biased lamellipodium development was lost within several minutes. The microtubule organizing center was located posterior to the nucleus in ECs under the influence of flow. However, this position was established only in ECs responding to fluid shear stress for longer than 1 h, indicating that positioning of the microtubule organizing center was not the reason for, but rather the result of, the biased lamellipodium response. Colcemid-treated ECs responded normally to flow, indicating that microtubules were not involved in both flow sensing and the flow-induced, biased lamellipodium development.
Masuda
M
M
Department of Structural Analysis, National Cardiovascular Center Research Institute, Osaka, Japan.
Fujiwara
K
K
D000818:Animals; D002417:Cattle; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003703:Demecolcine / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000648:ultrastructure*; D008870:Microtubules / Q000648:ultrastructure*; D013314:Stress, Mechanical
D003703:Demecolcine