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- W2368276257 abstract "Intermediate filaments(IFs) are prominent structural feature of the cytoskeleton of higher eukaryotic cells. A ariety of higher vertebrate cells of mesenchymal origin, such as fat tissue, have been shown to contain vimentin IF. The specific expression of vimentin IFs was maintained during adipose conversion. A better understanding of the significance of the disrupution of vimentin becomes important in characterizing the mechanism through which vimentin affects fat generation. The present experiments were carried out to examine the effects of anti-vimentin monoclonal antibody microinjected into cytoplasm on the proliferation and differentiation of rat preadiocyte and morphology of endogenous vimentin. Preadipocytes were isolated from adipose tissue of Sprague-Dawley rats, aged 20 days, by collagenase digestion. Cells were seeded at a density of 50-100 cells on each glass coverslip in M199 medium plus 10% fetal calf serum and cultured at 37℃ under a humidified atmosphere of 95% air:5% CO_(2). One day later, at the initiation of the differentiation, purified anti-vimentin McAb was microinjected into the cells, at a concentration of 1mg/ml, while setting cells of both human IgG microinjection and norr microinjection as control groups. To determine if disruption of the vimentin IF network would affect the capacity of predipocytes to accumulate lipid droplets during adipose conversion, the marker enzyme triglyceride(TG), as a specific and quantifiable marker for adipose conversion, was assayed spectrophotometrically. Final1y, at the indicated times following microinjection, the cells were processedfor anti-vimentin indirect immunofluorescence, to investigate the morphology alternation of the vimentin intermediate filament. The following characteristics of cells with anti-vimentin McAb microinjection were oberserved: (1)the cells were under apoptosis condition, hardly without lipid droplet in the cytoplasm, and nuclear fragmented into blue condensed particles or block detecting by Hoechst33342; (2) the total number of proliferating cells and the TG values were significantly lower than those of control group;(3) fluorescence staining of vimentin protein became weaker. Anti-vimentin McAb-induced depolymerization of the microtubule system leads to the collapse of extended vimentin IFs filament bundles into short agglomerations with the distributions of disperse disorder, while the vimentin in either non-microinjection or human IgG microinjection were localized to the surface of these lipid globules. These results suggest that the proliferation and differentiation of preadipocytes are obviously restrained by the microinjection of anti-vimentin McAb and vimentin network might be involved in the proliferation and differentiaion of preadipocytes." @default.
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- W2368276257 date "2003-01-01" @default.
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- W2368276257 title "Effects of cytoplasm microinjection of anti-vimentin monoclonal antibody on the prolifertion and differentiation of rat preadipocytes and morphology of vimentin" @default.
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