Figure 1: The GSH system is a major defense against Puma-dependent oxidative stress-induced apoptosis in developing cortical neurons.
From: Synaptic NMDA receptor activity is coupled to the transcriptional control of the glutathione system
(a,b) Inhibition of GCL activity with BSO treatment depletes cortical neurons of glutathione. Cortical neurons were treated with BSO (200 μM here and throughout) for 24 h, after which GSH levels were measured using in vivo labelling with MCB or a colourimetric assay of total glutathione in cell-free extracts (see methods). *P=0.0009 (a, n=5), 0.013 (b) n=4), Student’s t-test here and throughout unless otherwise stated. Exact P values are quoted throughout unless P<0.0001. Mean±s.e.m. is shown here and throughout. (c) Oxidative stress-induced Puma mRNA expression is potentiated by GSH depletion. Neurons were treated with BSO for 24 h, then subsequently treated with 50 or 100 μM H2O2 and Puma expression analysed by qRT–PCR, normalized to Gapdh. *P=0.024, 0.005 referring to asterisks as shown from left to right (here and throughout). One-way analysis of variance followed by Fisher’s post hoc test (1WA-Fph), n=4. (d) Neuronal vulnerability to H2O2-induced death is potentiated by GSH depletion. Neurons were treated with BSO for 24 h, then treated with different concentrations of H2O2, with cell death analysed 24 h later. *P<0.0001, <0.0001, 1WA-FPh; #P=0.0352 compared with control condition (n=4 (BSO), n=2 (Con)). (e) Example pictures from d. (f) H2O2-induced neuronal apoptosis is Puma dependent. Puma+/+ and Puma−/− cortical neurons were exposed to 100 μM H2O2. *P<0.0001, <0.0001; 2WA-FPh (n=6 Puma+/+; n=4 Puma−/−).
