Actin depolymerization and polymerization are required during apoptosis in endothelial cells.
In order to understand the role of actin microfilaments in the apoptotic process, we followed their evolution during tumor necrosis factor-alpha (TNF)-induced apoptosis in bovine aortic endothelial (BAE) cells. Using Western blotting analysis and immunofluorescence microscopy, we observed that the actin microfilaments network was disrupted in apoptotic cells. Depolymerization of F-actin was concomitant with internucleosomal DNA degradation and with the morphological changes associated with apoptotic cell death. However, using the actin microfilament disrupting agent, cytochalasin, we present evidence that the formation of blebs leading to apoptotic cell fragmentation requires neopolymerization of actin. Indeed, in the presence of cyochalasin, induction of apoptosis (internucleosomal DNA degradation) in BAE cells by TNF and cycloheximide was not associated with these classical morphological markers of apoptosis. Moreover, when added to BAE cells showing incipient apoptotic fragmentation, cytochalasin E reversed this process. We also observed an accumulation of actin at the basis of the apoptotic bodies in formation in these cells. Together, these results suggest that the actin network of flattened cells is disrupted concomitantly to the morphological modifications associated to the apoptotic cell death, and that the cytochalasin-sensitive reorganisation of actin is required to the formation of apoptotic blebs.
Suarez-Huerta
N
N
Institut de Recherche Interdisciplinaire en Biologie Humaine et Nucléaire (IRIBHN), Faculté de Médecine, Free University of Brussels (U.L.B.), Brussels, Belgium. nsuarez@ulb.ac.be
Mosselmans
R
R
Dumont
J E
JE
Robaye
B
B
D000199:Actins / Q000187:drug effects / Q000502:physiology*; D000818:Animals; D001011:Aorta / Q000187:drug effects; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D003572:Cytochalasins / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D006801:Humans; D008854:Microscopy, Electron; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000199:Actins; D003572:Cytochalasins; D014409:Tumor Necrosis Factor-alpha