Inhibitors of cation-chloride-cotransporters affect hypoxic/hypoglycemic injury in hippocampal slices.
Electroneutral cation-chloride cotransporters are abundantly expressed in the brain and are involved in the regulation of the intracellular Cl(-) concentration and thus gamma-aminobutyric acid-dependent inhibition of neuronal excitability. As yet there is little evidence whether or not Na(+)-K(+)-2Cl(-) or K(+)-Cl(-) cotransporters are involved in neuronal hyperexcitability and death in cerebral ischemia. In this study, by measuring propidium iodide staining in organotypic hippocampal slice cultures from young rats and population spike recovery in acutely isolated hippocampal slices from adult rats after a hypoxic/hypoglycemic insult, we were able to assess if cation-chloride cotransport inhibitors reduce neuronal injury. The Na(+)-K(+)-2Cl(-) cotransport inhibitor bumetanide in the range of 1-10 microM reduced neuronal damage in the slice cultures by 25%, but did not affect population spike recovery in acutely isolated slices. In contrast the K(+)-Cl(-) cotransport inhibitor [(dihydroindenyl)oxy] alkanoic acid (DIOA, 100 microM) significantly diminished the restitution of the population spikes from 33% before to 8% after hypoxia/hypoglycemia and increased the damage in the slice cultures by 60%. Consequently, our data suggest that the Na(+)-K(+)-2Cl(-) cotransporter may contribute to neuronal injury and that the activity of the K(+)-Cl(-) cotransporters is an intrinsic protective mechanism of neurons against ischemic damage.
Busse
Sebastian
S
Project Group Neuropharmacology, Leibniz Institute for Neurobiology, Germany.
Breder
Jörg
J
Dinkel
Klaus
K
Reymann
Klaus G
KG
Schröder
Ulrich H
UH
D000367:Age Factors; D000818:Animals; D002034:Bumetanide / Q000494:pharmacology; D002264:Carboxylic Acids / Q000494:pharmacology; D027682:Cation Transport Proteins / Q000187:drug effects / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D004232:Diuretics / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D007003:Hypoglycemia / Q000378:metabolism* / Q000473:pathology; D020925:Hypoxia-Ischemia, Brain / Q000378:metabolism* / Q000473:pathology; D066298:In Vitro Techniques; D007192:Indenes / Q000494:pharmacology; D017136:Ion Transport; D008297:Male; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D018696:Neuroprotective Agents / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D049994:Sodium Potassium Chloride Symporter Inhibitors; D028021:Sodium-Potassium-Chloride Symporters / Q000187:drug effects / Q000378:metabolism*
C059990:((dihydroindenyl)oxy)alkanoic acid; D002264:Carboxylic Acids; D027682:Cation Transport Proteins; D004232:Diuretics; D007192:Indenes; D018696:Neuroprotective Agents; D049994:Sodium Potassium Chloride Symporter Inhibitors; D028021:Sodium-Potassium-Chloride Symporters; D002034:Bumetanide