Apoptosis in human embryo development: 3. Fas-induced apoptosis in brain primary cultures |
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Authors: | Nat R Radu E Regalia T Popescu L M |
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Institution: | Division of Cellular and Molecular Medicine, "Carol Davila" University of Medicine and Pharmacy, Bucharest, Romania |
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Abstract: | Fas (APO-1/CD95) is an important apoptotic mediator for both immune and nervous systems. In the present study, we have investigated the expression and function of Fas in human embryonic/fetal brain primary cultures from 12 human embryos and fetuses with gestational ages between 5 to 22 weeks. Anti-Fas fluorescent antibody was used for labeling of Fas positive cells and for quantitation of Fas expression in brain cultures. To demonstrate that Fas receptor is functional in human embryonic/fetal brain cells, anti-Human-Fas monoclonal antibody (0.5 μg/ml) was used to induce apoptosis in brain primary cultures. Apoptosis was investigated by flow-cytometry and fluorescent microscopy using TUNEL and annexin V labeling. Fas was found to be expressed in the embryonic/fetal human primary brain cultures, on neuronal and glial cells or their precursors, varying with gestational ages. Cross-linking of Fas induced apoptosis in brain cultures indicating that Fas receptor functions as a death receptor. We also showed that cell death triggered through Fas receptor was caspase dependent, hence it was blocked by a selective caspase-8 inhibitor (IETD-fmk).These results suggest that Fas is involved in neuronal apoptosis in the developing human brain. |
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Keywords: | Fas (APO-1/CD95) apoptosis human development brain primary culture -TUNEL annexin V caspase-8 neurons glial cells |
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