Cytoarchitecture of Kirsten sarcoma virus-transformed rat kidney fibroblasts: butyrate-induced reorganization within the actin microfilament network.
Murine sarcoma virus-transformed rat fibroblasts (KNRK cells) undergo marked cytoarchitectural reorganization during in vitro exposure to sodium-n-butyrate (NaB) resulting in restoration of (1) a more typical fibroblastoid morphology, (2) proper cell-to-cell orientation, and (3) substratum adherence. Augmented cell spreading, involving greater than 90% of the population, was a function of culture density and time of exposure to NaB (2 mM final concentration). Induced cell spreading reflected a 2.5- to 3.0-fold increase in both total cellular actin content and deposition of actin into the detergent-resistant cytoskeleton. Cytoskeletal actin deposition in response to NaB was accompanied by the formation of occasionally dense, parallel alignments of F-actin-containing microfilaments and by a dramatic increase in the size and incidence of actin-enriched membrane ruffles. Long-term NaB-treated cells exhibited parallel orientations of microfilaments similar to those found in untransformed fibroblasts. Increased cytoskeletal actin occurred within 24 hr of NaB exposure, correlating with the initial reorganization of actin-containing microfilaments detected microscopically, and reflected concomitant 3-fold increases in cellular alpha-actinin and fibronectin content. In contrast, the amount of vimentin, tropomyosin, and tubulin in NaB-treated cells was significantly decreased. NaB-induced morphologic restructuring of sarcoma virus-transformed fibroblasts, thus, impacts on all three basic cytoskeletal systems. Selective increases, however, were evident in particular cytoskeletal proteins (actin, alpha-actinin, fibronectin) implicated in microfilament networking and cell spreading.
Ryan
M P
MP
Laboratory of Cell and Molecular Biology, Veterans Administration Medical Center, Albany, New York 12208.
Higgins
P J
PJ
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D000199:Actins / Q000378:metabolism; D000818:Animals; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D002448:Cell Adhesion; D002450:Cell Communication; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D007708:Kirsten murine sarcoma virus / Q000502:physiology*; D008856:Microscopy, Fluorescence; D051381:Rats; D009053:Sarcoma Viruses, Murine / Q000502:physiology*
D000199:Actins; D002087:Butyrates; D020148:Butyric Acid