title: Microfilament-disrupting agent latrunculin A induces and increased number of fenestrae in rat liver sinusoidal endothelial cells: comparison with cytochalasin B.
abstract: This report describes the effect of the actin-disrupting marine toxin latrunculin A on the cytoskeleton and fenestrae of liver endothelial cells (LECs). Fluorescence microscopy and whole mount-transmission electron microscopic preparations of isolated, purified, and cultured LECs showed that latrunculin A, which sequesters actin monomers and depolymerizes actin filaments, caused profound changes in microfilament organization in LECs. Scanning electron microscopic preparations showed that latrunculin A almost doubles the number of fenestrae within 10 minutes, whereas the diameter is only slightly reduced. All new fenestrae possess the earlier described fenestrae-associated cytoskeleton ring. Cytochalasin B, which disrupts the network of actin filaments, principally by capping the fast growing end of actin filaments, produced comparable effects with regard to actin organization and the number and size of fenestrae. After 1 hour of treatment, an equal maximum number of fenestrae was observed for both agents. The effect of latrunculin A was obtained at concentrations about 100 times lower than cytochalasin B. Thus, two agents that alter the state of actin organization in LECs, albeit by different mechanisms, cause the doubling of the number of fenestrae within 10 to 30 minutes. This indicates that the state of assembly of the actin cytoskeleton is important in the numerical dynamics of LEC fenestrae and that the actin cytoskeleton of LECs is probably the main mechanical regulator for sieving between the sinusoidal blood and the parenchymal cells. Latrunculin A represents a new agent in the study of the de novo formation of fenestrae.
authors.affiliation: Laboratory for Cell Biology and Histology (VUB), Laarbeklaan, Brussels-Jette, Belgium.
authors.forename: F
authors.initials: F
authors.lastname: Braet
authors.forename: R
authors.initials: R
authors.lastname: De Zanger
authors.forename: D
authors.initials: D
authors.lastname: Jans
authors.forename: I
authors.initials: I
authors.lastname: Spector
authors.forename: E
authors.initials: E
authors.lastname: Wisse
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects*; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology*; D002478:Cells, Cultured; D003571:Cytochalasin B / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D008102:Liver Circulation / Q000187:drug effects*; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D011161:Porifera; D051381:Rats; D013844:Thiazoles / Q000494:pharmacology*; D053778:Thiazolidines
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D013844:Thiazoles; D053778:Thiazolidines; D003571:Cytochalasin B; C037067:latrunculin A