Stimulated efflux of amino acids and glutathione from cultured hippocampal slices by omission of extracellular calcium: likely involvement of connexin hemichannels.
Omission of extracellular Ca(2+) for 15 min from the incubation medium of cultured hippocampal slices stimulated the efflux of glutathione, phosphoethanolamine, hypotaurine, and taurine. The efflux was reduced by several blockers of gap junctions, i.e. carbenoxolone, flufenamic acid, and endothelin-1, and by the connexin43 hemichannel blocking peptide Gap26 but was unchanged by the P2X(7) receptor inhibitor oxidized ATP, a pannexin1 hemichannel blocking peptide and an inactive analogue of carbenoxolone. Pretreatment of the slices with the neurotoxin N-methyl-d -aspartate left the efflux by Ca(2+) omission unchanged, indicating that the stimulated efflux primarily originated from glia. Elevated glutamate efflux was detected when Ca(2+) omission was combined with the glutamate uptake blocker l-trans-pyrrolidine-2,4-dicarboxylate and when both Ca(2+) and Mg(2+) were omitted from the medium. Omission of Ca(2+) for 15 min alone did not induce delayed toxicity, but in combination with blocked glutamate uptake, significant cell death was observed 24 h later. Our results indicate that omission of extracellular Ca(2+) stimulates efflux of glutathione and specific amino acids including glutamate via opening of glial hemichannels. This type of efflux may have protective functions via glutathione efflux but can aggravate toxicity in situations when glutamate reuptake is impaired, such as following a stroke.
Stridh
Malin H
MH
Institute of Neuroscience and Physiology, University of Gothenburg, SE-40530 Gothenburg, Sweden.
Tranberg
Mattias
M
Weber
Stephen G
SG
Blomstrand
Fredrik
F
Sandberg
Mats
M
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D017630:Connexins / Q000378:metabolism*; D017629:Gap Junctions / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D006624:Hippocampus / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D010455:Peptides / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D000596:Amino Acids; D017630:Connexins; D005971:Glutamates; D010455:Peptides; D005978:Glutathione; D002118:Calcium