Fig. 10

Vacuolation of cytoplasm and cytoskeletal modifications. Phase contrast (a), TEM (b, c, e, f), and acridine orange (AO) fluorescence (d) micrographs of PEG-treated NIHs cells. Images in a, c, e, and f refer to cells subsequently incubated with vinblastine. In a and b, signs of cytoplasmic changes in polyploid/aneuploid cells are visible as granulations (phase contrast micrograph of a multinucleate cell in a) and vacuolations (TEM micrograph in b). In c, a detail of the vacuoles that appear to be empty or with variable content. AO staining (d) allowed us to observe dark areas (arrows) in the cytoplasm of a polyploid/aneuploid cell. This aspect may be related to the images obtained by TEM microscopy, which allowed us to note electron-transparent areas of cytoplasmic discontinuity (arrows), especially in VBL-incubated cells (e). In e and f, VBL effects on the cytoskeleton can also be observed: in particular, in e, tubulin paracrystals (arrowheads) close to areas of extensive vacuolation (arrows) are visible. In f, a bundle of intermediate filaments (arrowhead) in proximity to a vacuolation area is evident. Vacuoles show heterogeneous content, which may result from the degradation of organelles or endomembranes (myelin figures indicated by arrows). Scale bars in a and d: 10 µm; in b: 1 µm; in c and e: 3 µm; in f: 1 µm