RESUMO
In a gradient of chemoattractant, Dictyostelium cells are orientated with their front directed toward the source and their tail pointing into the opposite direction. The front region is specified by the polymerization of actin and the tail by the recruitment of filamentous myosin-II. We have dissected these front and tail responses by exposing cells to an upshift of cyclic AMP. A sharp rise and fall of polymerized actin within 10s is accompanied by the recruitment of proteins involved in turning actin polymerization on or off. The cortical accumulation of myosin-II starts when the front response has declined, supporting the concept of divergent signal transmission and adaptation pathways.
Assuntos
Polaridade Celular/efeitos dos fármacos , Fatores Quimiotáticos/farmacologia , Dictyostelium/citologia , Dictyostelium/efeitos dos fármacos , 4-Butirolactona/análogos & derivados , 4-Butirolactona/metabolismo , Actinas/metabolismo , Animais , AMP Cíclico/farmacologia , Miosinas/metabolismo , Transporte Proteico/efeitos dos fármacos , Proteínas de Protozoários/metabolismo , Fatores de TempoRESUMO
The preferentially expressed antigen in melanoma (PRAME) gene is aberrantly expressed in chronic lymphoproliferative disorders (CLD). We produced and characterized an anti-PRAME monoclonal antibody (MoAb), which was then applied in a quantitative flow cytometric (QFC) method to evaluate PRAME expression in leukemic cells from the peripheral blood (PB) of 47 patients with chronic lymphocytic leukemia and seven with mantle cell lymphoma as well as in the PB mononuclear cells (PBMCs) and B lymphocytes from 15 healthy subjects. Approximately 90% of CLD, but none of the normal samples, presented more than 20% of PRAME+ lymphocytes. Moreover, the intensity of PRAME expression was significantly higher in CLD cells compared to normal B lymphocytes and PBMCs. By immunofluorescence microscopy and by permeabilized flow cytometry we demonstrated that PRAME is a membrane antigen and a cytoplasmic protein aberrantly expressed in malignant CLD. Our results suggest that the analysis of PRAME protein may contribute for the distinction between normal and leukemic cells in CLD, and that PRAME may be a potential target for therapy.