Expression of the gene of the alpha-smooth muscle-actin isoform in rat liver and in rat fat-storing (ITO) cells

Abstract

Fat storing cells (FSCs) in the liver represent the main site of vitamin A deposition in the body. These cells are considered to play an important role during scar formation and fibrogenesis in the liver. The putative descent of FSCs from the fibroblastic or from the myofibroblastic system have not been determined yet by morphological or immunohistochemical studies. To further define the origin of these liver cells, we analysed the pattern of expression of three structural proteins: vimentin, desmin and the alpha-smooth muscle (SM)-actin isoform in FSCs of the rat liver, in smooth muscle cells (SMCs) from the aorta and in rat skin fibroblasts. FSCs were studied by immunohistochemical methods immediately after isolation, at days 3 and 7 after plating. FSC-gene-expression was also analysed by Northern blot analysis of total RNA extracted from cells in culture at days 3 and 7 after isolation. Arterial SMCs and skin fibroblasts were studied in a similar way. For comparison, isolated rat hepatocytes and Küpffer cells (Kc) were studied. Of freshly isolated FSCs, 100% were vimentin-positive, 50% were desmin-positive, but all were alpha-SM-actin negative. Three days after isolation, FSCs were clearly positive for vimentin and desmin and weakly alpha-SM-actin-positive, as demonstrated by indirect immunofluorescence as well as by the immunoperoxidase technique. Desmin, alpha-SM-actin and vimentin staining was further increased at day 7 after isolation, and alpha-actin specific transcripts in FSC-RNA were clearly detectable at day 7 after isolation. Passaged arterial SMCs were vimentin- and alpha-SM-actin-positive, but desmin-negative and fibroblasts were only vimentin-positive.(ABSTRACT TRUNCATED AT 250 WORDS)