Cytochalasin D and cationized ferritin as probes for the morphological investigation of blebbing in two human cell lines

William D. Meek, Walter L. Davis

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

The potent fungal metabolite cytochalasin D (CD) and cationized ferritin (CF) are used in combination to test for negative charge distribution on blebs (knobs). Two established human epithelial cell lines, WISH and HeLa, that display blebs in various phases of the cell cycle or under certain culture conditions (37,46) are investigated. CD alone, applied at a low concentration (1.0 μg/ml) and for a short time period (3 min), causes blebs to appear as the prevalent surface feature. These are filled mainly with free ribosomes. Additionally, feltlike mats, presumed to be disorganized, compacted microfilaments, are formed directly beneath the cell membrane. These are especially evident in the cortical cytoplasm below the blebs or bleb clusters. CF (0.345 mg/ml), applied for a 5-min period after CD administration (1.0 μg/ml) for 3 min, appears along the surface of microvilli, at the base of blebs, and in vesicles beneath the bleb clusters. In some cases, microfilaments (6 nm in diameter) are closely related to the vesicles. CF does not preferentially bind to the apical cell membrane of blebs. Above areas of the subplasmalemmal microfilaments, CF membrane binding is apparent, even under circumstances where the filaments are disorganized by cytochalasin treatment. These results seem to show the following: (a) bleb membranes are different from the remainder of the cell and do exhibit a loss of negative charge and (b) surface charge may be dependent on the presence or structural integrity of membrane-related 6-nm microfilaments.

Original languageEnglish
Pages (from-to)725-737
Number of pages13
JournalIn Vitro Cellular & Developmental Biology
Volume22
Issue number12
DOIs
StatePublished - 1 Dec 1986
Externally publishedYes

Keywords

  • blebbing
  • cationized ferritin
  • cytochalasin D
  • HeLa
  • microfilaments
  • tissue culture
  • transmission electron microscopy
  • WISH

Fingerprint Dive into the research topics of 'Cytochalasin D and cationized ferritin as probes for the morphological investigation of blebbing in two human cell lines'. Together they form a unique fingerprint.

  • Cite this