Actions of cytochalasins on the organization of actin filaments and microtubules in a neuronal growth cone.
Open Access
- 1 October 1988
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 107 (4), 1505-1516
- https://doi.org/10.1083/jcb.107.4.1505
Abstract
Actions of cytochalasin B (CB) on cytoskeletons and motility of growth cones from cultured Aplysia neurons were studied using a rapid flow perfusion chamber and digital video light microscopy. Living growth cones were observed using differential interference contrast optics and were also fixed at various time points to assay actin filament (F-actin) and microtubule distributions. Treatment with CB reversibly blocked motility and eliminated most of the phalloidin-stainable F-actin from the leading lamella. The loss of F-actin was nearly complete within 2-3 min of CB application and was largely reversed within 5-6 min of CB removal. The loss and recovery of F-actin were found to occur with a very distinctive spatial organization. Within 20-30 s of CB application, F-actin networks receded from the entire peripheral margin of the lamella forming a band devoid of F-actin. This band widened as F-actin receded at rates of 3-6 microns/min. Upon removal of CB, F-actin began to reappear within 20-30 s. The initial reappearance of F-actin took two forms: a coarse isotropic matrix of F-actin bundles throughout the lamella, and a denser matrix along the peripheral margin. The denser peripheral matrix then expanded in width, extending centrally to replace the coarse matrix at rates again between 3-6 microns/min. These results suggest that actin normally polymerizes at the leading edge and then flows rearward at a rate between 3-6 microns/min. CB treatment was also observed to alter the distribution of microtubules, assayed by antitubulin antibody staining. Normally, microtubules are restricted to the neurite shaft and a central growth cone domain. Within approximately 5 min after CB application, however, microtubules began extending into the lamellar region, often reaching the peripheral margin. Upon removal of CB, the microtubules were restored to their former central localization. The timing of these microtubule redistributions is consistent with their being secondary to effects of CB on lamellar F-actin.Keywords
This publication has 45 references indexed in Scilit:
- Subcellular compartmentalization by local differentiation of cytoplasmic structureCell Motility, 1988
- Distribution of actin and tubulin in cells and in glycerinated cell models after treatment with cytochalasin B (CB)Experimental Cell Research, 1976
- Cytochalasin B inhibits actin-related gelation of HeLa cell extracts.The Journal of cell biology, 1976
- A new hypothesis of contact guidance in tissue cellsExperimental Cell Research, 1976
- Organization of an actin filament-membrane complex. Filament polarity and membrane attachment in the microvilli of intestinal epithelial cells.The Journal of cell biology, 1975
- Spectroscopic evidence for the interaction of phalloidin with actinFEBS Letters, 1975
- Biochemical studies on the mode of action of cytochalasin B. Cytochalasin B binding to red cell membrane in relation to glucose transport.1974
- ACTION OF CYTOCHALASIN D ON CELLS OF ESTABLISHED LINESThe Journal of cell biology, 1974
- ULTRASTRUCTURE AND FUNCTION OF GROWTH CONES AND AXONS OF CULTURED NERVE CELLSThe Journal of cell biology, 1971
- Microfilaments in Cellular and Developmental ProcessesScience, 1971