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Loss- and Gain-of-function Approach to Investigate Early Cell Fate Determinants in Preimplantation Mouse Embryos
Published on: June 6, 2016
IFN-gamma induces apoptosis in mouse embryonic stem cells, a putative mechanism of its embryotoxicity
G M Zou1, M F Reznikoff-Etievant, F Hirsch
1Department of Gynecology/Obstetrics, Saint-Antoine Hospital, France. zougm@francemultimedia.fr
Abstract:
It has been reported that interferon (IFN)-gamma should inhibit in vitro mouse embryo growth by direct cell toxicity. However, the mechanism involved has not been clearly established. In the present study, this question was addressed using the embryonic stem (ES) cell model. It was found that IFN-gamma, induces a dose-dependent apoptosis in ES cells, as assessed by trypan-blue staining, by Annexin-V labeling and DNA analysis, Moreover, IFN-gamma treatment cooperates with Fas-mediated apoptosis, a phenomenon that has been recently reported. As Bcl-2 oncoprotein functions as a death repressor molecule in an evolutionarily conserved cell death pathway, its expression was analyzed by flow cytometry. It was demonstrated that Bcl-2 is expressed in ES cells. When compared to untreated ES cells, IFN-gamma-treated, apoptotic cells expressed a lower Bcl-2 level and a normal level of Fas, whereas surviving cells expressed a normal level of Bcl-2 but a lower Fas expression. Altogether, these data suggest that IFN-gamma may influence early mouse embryo development by promoting apoptosis, which may constitute a novel mechanism of IFN-gamma embryotoxicity.
Insights
Interferon-gamma (IFN-γ) induces apoptosis in mouse embryonic stem cells, potentially explaining its embryotoxic effects. This study reveals IFN-γ
Area of Science:
- Developmental Biology
- Immunology
- Cell Biology
Background:
- Interferon-gamma (IFN-γ) is known to inhibit mouse embryo growth via direct cell toxicity.
- The precise mechanism underlying IFN-γ-induced embryotoxicity remains unclear.
- Embryonic stem (ES) cells serve as a model to investigate early developmental processes.
Purpose of the Study:
- To elucidate the mechanism by which IFN-γ affects mouse embryonic stem cells.
- To determine if IFN-γ induces apoptosis in ES cells and its relation to cell death pathways.
Main Methods:
- Treatment of mouse ES cells with varying doses of IFN-γ.
- Assessment of apoptosis using trypan-blue staining, Annexin-V labeling, and DNA analysis.
- Flow cytometry analysis of Bcl-2 and Fas expression in treated and untreated ES cells.
Main Results:
- IFN-γ induced a dose-dependent apoptosis in ES cells.
- IFN-γ treatment enhanced Fas-mediated apoptosis.
- Apoptotic ES cells showed decreased Bcl-2 and normal Fas levels, while surviving cells had normal Bcl-2 and decreased Fas.
Conclusions:
- IFN-γ promotes apoptosis in mouse embryonic stem cells.
- The observed changes in Bcl-2 and Fas expression suggest a role in IFN-γ-induced cell death.
- IFN-γ-induced apoptosis may represent a novel mechanism of its embryotoxic effects on early mouse development.
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