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Chelation of endogenous membrane calcium inhibits gamma-aminobutyric acid uptake in synaptosomes



Chelation of endogenous membrane calcium inhibits gamma-aminobutyric acid uptake in synaptosomes



Journal of Neuroscience Research 24(2): 293-298



In a previous work, we have demonstrated that calcium chelators induce the release of gamma-aminobutyric acid (GABA) from synaptosomes in a Na+ -dependent manner and that this release is blocked by cations such as Mg2+, La3+, and ruthenium red. In the present study, we show that treatment of synaptosomes with 0.1 mM EGTA in the absence of both Ca2+ and Mg2+ inhibits the sodium-dependent high-affinity uptake of [3H]GABA by about 50%. This inhibition increased to about 65% with 1.5 mM EGTA, and it was completely prevented by an excess of Ca2+ or by 1.2 mM Mg2+. In contrast, when EDTA was used as a chelator, Mg2+ was unable to reverse the inhibition. The inhibitory effect of 0.1 mM EGTA was also prevented by 250 microM La3+ or by 20 microM ruthenium red. In the absence of chelators and the presence of Ca2+ and Mg2+, 50 microM and 200 microM La3+ inhibited GABA uptake by about 20 and 50%, respectively, whereas 20 microM ruthenium red produced a nonsignificant 25% inhibition and nifedipine was without effect. It is concluded that the membrane-surface negative charges, probably those of the sialic acid molecules that have been implicated in the functioning of the GABA carrier, must be neutralized by endogenous Ca2+ or by another cation in order to permit the adequate function of the transporter. The inhibition by La3+ in the absence of the chelators could be explained by a binding of this cation to the Na+ sites on the GABA carrier.

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Accession: 039528130

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PMID: 2479766

DOI: 10.1002/jnr.490240222


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