NHE 1 in cerebral ischemic damage 1 ERK 1 / 2P 90 RSK-MEDIATED PHOSPHORYLATION OF NA + / H + EXCHANGER ISOFORM 1 : A ROLE IN ISCHEMIC NEURONAL DEATH

The function and regulation of Na/H exchanger isoform 1 (NHE1) following cerebral ischemia are not well understood. In this study, we demonstrate that extracellular signal related kinases (ERK1/2) play a role in stimulation of neuronal NHE1 following in vitro ischemia. NHE1 activity was significantly increased during 10-60 min reoxygenation (REOX) after 2 h oxygen and glucose deprivation (OGD). OGD/REOX not only increased the Vmax for NHE1 but also shifted the Km toward decreased [H]i. These changes in NHE1 kinetics were absent when MAPK/ERK kinase (MEK) was inhibited by the MEK inhibitor U0126. There were no changes in level of phosphorylated ERK1/2 (p-ERK1/2) after 2 h OGD. p-ERK1/2 level was significantly increased during 10-60 min REOX, which was accompanied by a nuclear translocation. U0126 abolished REOXinduced elevation and translocation of pERK1/2. We further examined the ERK-90 kDa ribosomal S6 kinase (p90 ) signaling pathways. At 10 min REOX, phosphorylated NHE1 was increased with a concurrent elevation of phosphorylation of p90, a known NHE1 kinase. Inhibition of MEK activity with U0126 abolished phosphorylation of both NHE1 and p90. Moreover, neuroprotection was observed with U0126 or genetic ablation or pharmacological inhibition of NHE1 following OGD/REOX. Taken together, these results suggest that activation of ERK1/2p90 pathways following in vitro ischemia phosphorylates NHE1 and increases its activity, which subsequently contributes to neuronal damage.