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. 2007 May 1;5(4):417–428. doi: 10.1111/j.1582-4934.2001.tb00177.x

Apoptosis in human embryo development: 3. Fas‐induced apoptosis in brian primary cultures

Roxana Nat 1, E Radu 1, T Regalia 1, L M Popescu 1,✉
PMCID: PMC6740267  PMID: 12067476

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.

Keywords: Fas (APO‐1/CD95), apoptosis, human development, brain primary culture ‐TUNEL, annexin V, caspase‐8, neurons, glial cells

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