Cytoplasmic desmosomes and intermediate filament disturbance following acrylamide treatment in cultured rat keratinocytes

Abstract

The present paper describes disturbances in the organization of tonofilaments and desmosomes of rat lingual and epidermal keratinocytes after treatment of the cells with acrylamide in culture. This treatment induced changes in cell shape, reduction of intercellular adhesion and a perinuclear accumulation of cytoplasmic organelles. Using specific antibodies for cytokeratins, the filaments were disorganized particularly in the perinuclear region. In untreated cells, keratin filament labelling was very weak or absent above and below the nucleus thus leaving a black nuclear space in fluorescine microscopy. Following acrylamide treatment, the keratin filament labelling covered the nuclear space which indicated the accumulation of these filaments all around the nucleus. Furthermore, the desmosomal junctions were often associated with thick keratin bundles. Antibodies for desmoplakins revealed a reduction in intercellular labelling and stronger cytoplasmic labelling. Ultrastructurally, well-developed long tonofilaments were found to associate with large desmosomal junctions. Furthermore, small-sized desmosomal structures were identified within the cytoplasm. Morphologically, these were identical to cell surface desmosomes and were almost always associated with well-developed tonofilaments. The effect of acrylamide on the protein kinase A activity might be implicated in the disturbances of the desmosome-intermediate filament complex and in the initiation of contractile forces necessary for perinuclear accumulation of intermediate filaments and for the formation of intact cytoplasmic desmosomes. The acrylamide-induced intermediate filament and desmosomal changes may provide valuable information on the mechanism of intact cytoplasmic desmosome formation in several skin diseases and in squamous cell carcinoma.