The Chloride Transporter Na+-K+-Cl− Cotransporter Isoform-1 Contributes to Intracellular Chloride Increases after In Vitro Ischemia

Ischemic episodes in the CNS cause significant disturbances in neuronal ionic homeostasis. To directly measure changes in intracellular Cl− concentration ([Cl−]i) during and after ischemia, we used Clomeleon, a novel ratiometric optical indicator for Cl−. Hippocampal slices from adult transgenic mice expressing Clomeleon in hippocampal neurons were subjected to 8 min of oxygen-glucose deprivation (OGD) (an in vitro model for ischemia) and reoxygenated in the presence of glucose. This produced mild neuronal damage 3 h later that was prevented when the extracellular [Cl−] was maintained at 10 mm during reoxygenation. OGD induced a transient decrease in fluorescence resonance energy transfer within Clomeleon, indicating an increase in [Cl−]i. During reoxygenation, there was a partial recovery in [Cl−]i, but [Cl−]i rose again 45 min later. To investigate sources of Cl− accumulation, we examined the effects of Cl− transport inhibitors on the rises in [Cl−]i during and after OGD. Bumetanide and furosemide, which inhibit Cl− influx through the Na+-K+-Cl− cotransporter isoform-1 (NKCC-1) and efflux through the K+-Cl− cotransporter isoform-2, were unable to inhibit the first rise in [Cl−]i, yet entirely prevented the secondary rise in [Cl−]i during reoxygenation. In contrast, picrotoxin, which blocks the GABA-gated Cl− channel, did not inhibit the secondary rise in [Cl−]i after OGD. [Cl−]i increases during reoxygenation were accompanied by an increase in phosphorylation of NKCC-1, an indication of increased NKCC-1 activity after OGD. We conclude that NKCC-1 plays an important role in OGD-induced Cl− accumulation and subsequent neuronal damage.

• Publication year

  2006

• Venue

  Journal of Neuroscience

• Publication date

  2006-02-01

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1523/JNEUROSCI.1421-05.2006PMID 16452663PMCID PMC6675477
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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