Hypoglycemic neuronal death is triggered by glucose reperfusion and activation of neuronal NADPH oxidase
J. Clin. Invest. Sang Won Suh, et al. 117:910 doi:10.1172/JCI30077 [
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Figure 4Zinc chelation prevents activation of neuronal NADPH oxidase. (
A) FluoZin-3 images of intracellular free zinc levels in cultured neurons after 3 hours of GD or 2 hours of GD and 1 hour of glucose replacement (GD/GR). Fluorescence induced by GR was attenuated in cultures treated with CaEDTA during the GD/GR incubations and eliminated by 10-minute incubation with N,N,N',N'-tetrakis(2-pyridyl-methyl)ethylenediamine (TPEN).
n = 4. (
B) Immunostaining for the p47
phox and p67
phox subunits of NADPH oxidase in cultured neurons. GD/GR-induced migration of these subunits to the neuronal plasma membrane was blocked by the zinc chelator CaEDTA but not by ZnEDTA used as a control. Scale bar: 10 μm. Representative of 3 cultures under each condition. (
C) Vesicular zinc in the rat hippocampal hilus is imaged with TSQ fluorescence (white). The TSQ signal loss was greater after 30 minutes HG and 30 minutes GR (HG/GR) than after 60 minutes HG without GR. Fluorescence intensity is expressed as arbitrary units, with subtraction of background measured in the lateral ventricle (white square). Scale bar: 200 μm. Data are mean + SEM;
n = 10; *
P < 0.05; **
P < 0.01. (
D) TSQ fluorescence was increased in the postsynaptic pyramidal cells after HG/GR, and this increase was blocked by CaEDTA (
n = 3). Scale bar: 100 μm. (
E) CaEDTA reduces GR-induced superoxide production in the CA1 neurons. ZnEDTA was the control. Graph shows quantified CA1 neuronal Et fluorescence intensity in rats treated with intracerebroventricular saline, CaEDTA, or ZnEDTA. Scale bar: 100 μm. Data are mean + SEM;
n = 3–5; *
P < 0.05.
