The aims of the current study were to 1) examine the effects of hypoxia and acidosis on cultured cortical neurons, and 2) explore the role of transporters and ion channels in hypoxic injury. Cell injury was measured in cultured neurons or hippocampal slices following hypoxia (1% O₂) or acidosis (medium pH 6.8) treatment. Inhibitors of transporters and ion channels were employed to investigate their roles in hypoxic injury. Our results showed that (1) neuronal damage was apparent at 5-7 days of hypoxia exposure, i.e. 36-41% of total LDH was released to medium, (2) acidosis alone did not lead to significant injury compared to non-acidic, normoxic controls. Pharmacological studies revealed that (1) no significant difference in neuronal injury between controls and inhibition of Na⁺-K⁺ ATP pump, or voltage-gated Na⁺ channel, or K_ATP channel, or reverse mode of Na⁺-Ca²⁺ exchanger under hypoxia; however, (2) inhibition of HCO₃^- transporters with 500µM DIDS did not cause hypoxic death in both cultured cortical neurons and hippocampal slices; (3) in contrast, inhibition of Na⁺-H⁺ exchanger isoform 1 (NHE1) with either 10µM HOE642 or 2µM T-162559 resulted in dramatic hypoxic injury (+95% for HOE642 and +100% for T-162559 relative to normoxic control, p<0.001) on the treatment day 3, when no death occurred for hypoxic controls (no inhibitor). No further damage was observed by NHE1 inhibition on the treatment day 5. We conclude that inhibition of NHE1 accelerates hypoxia-induced neuronal damage. In contrast, DIDS rescues neuronal death under hypoxia. Hence, DIDS-sensitive mechanism may be a potential therapeutic target.