Effects of anisosmotic conditions on the cytoskeletal architecture of cultured PC12 cells.

Abstract

PC12 cells show a classical volume regulatory process when submitted to hypo-osmotic conditions. The present study examined the effects of such osmotic shock on the structural organization of different cytoskeletal elements. Results were obtained by use of different light and electron microscopy techniques combined with immunostaining methods. It appeared that the osmotically induced changes in cell volume were concomitant with important modifications in the organization of the microfilament network. Microfilaments concentrated in the perinuclear area, leaving only radial extensions of poorly organized structures in the cytoplasm. The latter were the only actin structures immunologically stained in the cytoplasm and seemed to anchor to the plasma membrane. Measurements of the fluorescence intensity of PC12 cells treated with FITC-labeled phalloidin indicated a progressive depolymerization, followed by a repolymerization of F-actin. This occurs in parallel with microfilament reorganization and volume regulatory processes. The appearance of microfilament reorganization was a function of both the incubation period and the amplitude of the osmolarity changes. During the first minutes of osmotic shock, a decrease was observed in the density and length of microvilli, which normally cover the PC12 cell surfaces, suggesting an early reorganization of the underlying microfilament network. Microtubules and intermediate filament networks were not affected by the hypo-osmotic conditions.