[Reorganization of microtubule system in pulmonary endothelial cells in response to thrombin treatment].
Thrombin induces rapid and reversible increase of endothelial (EC) barrier permeability associated with actin cytoskeleton remodeling and contraction. The role of microtubules (Mts) in EC barrier regulation compared with actin systems is poorly understood. In this work we studied pathways of Mt and actin regulation in response to thrombin treatment in cultured EC, and the involvement of trimeric G-proteins and in this process. Cells were treated with thrombin, and further analysed using immunofluorescent staining of actin and Mts, digital microscopy and morphometric analysis. In normal cells actin network consists of thin bundles basically located in the cell periphery, Mt density decreases from the cell center to the cell edge. Thrombin (25 nM) induced endothelial dysfunction associated with a rapid (within 5 min) decrease of peripheral Mt network and a slower actin stress fiber formation in the cytoplasm. Pretreatment with Pertussis toxin, which is Gi protein inhibitor, attenuated thrombin-induced stress fiber formation and Mt disassembly. Overexpression of activated G12, G13, Gi and Gq proteins, which are involved in thrombin receptor-mediated signaling, resulted in increasing stress fibers thickness and density and complete Mt disassembly. From the results obtained we suggest that thrombin regulates actin cytoskeleton of EC using local Mt depolymerization at the cell edge.
Smurova
K M
KM
Biriukova
A A
AA
Garcia
J G
JG
Vorob'ev
I A
IA
Alieva
I B
IB
Verin
A D
AD
D000199:Actins / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D044385:GTP-Binding Protein alpha Subunits / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000098:blood supply*; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D022502:Stress Fibers / Q000648:ultrastructure; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000494:pharmacology* / Q000502:physiology
D000199:Actins; D044385:GTP-Binding Protein alpha Subunits; D013917:Thrombin