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Am J Physiol Cell Physiol 281: C1205-C1210, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 4, C1205-C1210, October 2001

Role of Na+/H+ exchanger during O2 deprivation in mouse CA1 neurons

Hang Yao1, Xiang-Qun Gu1, Robert M. Douglas1, and Gabriel G. Haddad1,2

1 Section of Respiratory Medicine, Department of Pediatrics, and 2 Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520

To determine the role of membrane transporters in intracellular pH (pHi) regulation under conditions of low microenvironmental O2, we monitored pHi in isolated single CA1 neurons using the fluorescent indicator carboxyseminaphthorhodafluor-1 and confocal microscopy. After total O2 deprivation or anoxia (PO2 congruent  0 Torr), a large increase in pHi was seen in CA1 neurons in HEPES buffer, but a drop in pHi, albeit small, was observed in the presence of HCO<UP><SUB>3</SUB><SUP>−</SUP></UP>. Ionic substitution and pharmacological experiments showed that the large anoxia-induced pHi increase in HEPES buffer was totally Na+ dependent and was blocked by HOE-694, strongly suggesting the activation of the Na+/H+ exchanger (NHE). Also, this pHi increase in HEPES buffer was significantly smaller in Na+/H+ exchanger isoform 1 (NHE1) null mutant CA1 neurons than in wild-type neurons, demonstrating that NHE1 is responsible for part of the pHi increase following anoxia. Both chelerythrine and H-89 partly blocked, and H-7 totally eliminated, this anoxia-induced pHi increase in the absence of HCO<UP><SUB>3</SUB><SUP>−</SUP></UP>. We conclude that 1) O2 deprivation activates Na+/H+ exchange by enhancing protein kinase activity and 2) membrane proteins, such as NHE, actively participate in regulating pHi during low-O2 states in neurons.

hippocampus; transporter; anoxia; pH; sodium-hydrogen exchanger


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