The effect of oxygen tension on intracellular survival signalling in primary villous trophoblasts.

Villous trophoblasts undergo increased apoptosis and experience a wider gradient of oxygen tensions (pO(2)) in pregnancies complicated by intrauterine growth restriction. We hypothesize that pO(2)affects trophoblast apoptosis by altering survival signalling through the phosphatidylinositol-3 (PI-3)-kinase and mitogen activated protein kinase (MAPK) pathways. Cytotrophoblasts were cultured at pO(2)from <10 to approximately 140 mmHg with Epidermal Growth Factor (EGF), Vascular Endothelial Growth Factor (VEGF) and basic Fibroblast Growth Factor (bFGF) at concentrations of 0.1 to 10 ng/ml for 1 to 12 h, then assessed for apoptosis (TUNEL) and specific protein expressions (Western blot analysis). Spontaneous apoptosis was highest at <10 mmHg and lowest approximately 15 mmHg. Only EGF activated either signalling pathway at any pO(2). Inhibition of both pathways was required to inhibit EGF-stimulated survival. Maximal EGF activation of either pathway was insensitive to pO(2). At lower oxygen tensions, MAPK phosphorylation was maximal at 1 ng/ml EGF compared with 10 ng/ml for the PI-3-kinase path. The EGF receptor was spontaneously phosphorylated with increasing culture times at lower oxygen levels, an effect reflected down-stream by PI-3-kinase and Akt phosphorylation. We conclude that strong survival signalling in trophoblasts requires both PI-3- and MAP-kinase pathways, is rather insensitive to pO(2)changes and is spontaneously activated with increasing hypoxic exposure.