Effect of retinoic acid on the actin cytoskeleton in HL-60 cells.
The cytoskeleton, in addition to its structural and kinetic functions, is also involved in modulating signal transfer in cell proliferation, differentiation and death. In some myeloid leukemic cell lines, the process of cell differentiation accompanied by apoptosis, can be induced by all-trans retinoic acid (ATRA). In this report, we describe the morphological changes in actin cytoskeleton, taking place during apoptosis in cells of the human leukemic HL-60 cell line. By using fluorescent microscopy, the morphology of microfilaments and the proportion of apoptotic cells in the cell populations untreated or treated with 10(-6) M ATRA were detected. Interphase HL-60 cells showed aggregations of short, thick microfilament bundles in the region between the plasma membrane and the nucleus. In comparison with both interphase and mitotic cells, the cells with apoptotic nuclear fragmentation showed a different organisation of the actin cytoskeleton. The following types of F-actin structures were observed: (i) Cells with a high number of large dots/patches of F-actin under the plasma membrane. These dots might be localised only in the part of the cell or occurred under the whole plasma membrane. This arrangement was often associated with a diffuse signal for F-actin. (ii) Cells with 3D-network of F-actin fibres through the cytoplasm between remnants of the cell nucleus. This 3D-structure probably played an important active role in the process of apoptotic bodies formation. (iii) Cells without any detectable signal for F-actin or cells with only a very low F-actin signal. Both of these showed typical apoptotic collapse of chromatin. It is concluded that the actin cytoskeleton is a dynamic structure actively involved in the executive phase of the process of apoptosis. It is suggested that the rearrangement of the microfilament network and its subsequent degradation are necessary for the main morphological changes of apoptotic cells, i.e., plasma membrane blebbing and apoptotic bodies formation.
Zitterbart
K
K
Department of Biology, Faculty of Medicine, Masaryk University Brno, Czech Republic. kzitter@med.muni.cz
Veselská
R
R
D000199:Actins / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D003599:Cytoskeleton / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects; D008856:Microscopy, Fluorescence; D022423:Myeloid Cells / Q000166:cytology / Q000187:drug effects; D013997:Time Factors; D014212:Tretinoin / Q000494:pharmacology*
D000199:Actins; D014212:Tretinoin