In situ and In vitro Comparisons of Endothelial Cell G- and F-Actin Distribution
Abstract
Numerous studies have described the F-actin cytoskeleton, however, little information relevant to G-actin is available. Using in situ and in vitro conditions and rhodamine-conjugated probes specific for G- (deoxyribonuclease 1. 03 micromole) or F-actin (phalloidin, 0.2 micromoles), the actin pools of bovine aortic endothelial cells were examined. Cells in situ displayed a diffuse g- actin distribution, while F-actin was concentrated in the ectoplasm and in fine stress fibers that traversed some cells. Cells of sub-confluent or just confluent in vitro cultures demonstrated intense fluorescence, with many F-actin stress fibers. Post confluent cultures resembled in in situ condition, since peripheral F-actin was prominent, traversing actin stress fibers were greatly reduced and fluorescent intensity was diminished. Post confluency had little influence on G-actin, with only and enhancement in the intensity of G-actin punctate fluorescence. When post confluent cultures were treated with cytochalasin-D, F-actin networks were disrupted and F-actin punctate and diffuse fluorescence increased. G-actin fluorescence was not altered by this disruption. Though its unstructured nature may account for the minor changes observed, the stability of the G-actin pool in the presence of notable F-actin modulations suggested that filamentous actin was the key constituent involved in these actin cytoskeletal alterations.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 1989
- Accession Number
- ADA206247
Entities
People
- David A. Dubose
- Jose A. Guzman
- Robert J. Carpenter Jr
- Rosaria Haugland
Organizations
- United States Army Research Institute of Environmental Medicine