Nigericin-induced Na+/H+ and K+/H+ exchange in synaptosomes: effect on [3H]GABA release

The effect of the putative K+/H+ ionophore, nigericin on the internal Na+ concentration ([Nai]), the internal pH (pHi), the internal Ca2+ concentration ([Cai]) and the baseline release of the neurotransmitter, GABA was investigated in Na+-binding benzofuran isophtalate acetoxymethyl ester (SBFI-AM), 2',7'-bis(carboxyethyl)-5(6) carboxyfluorescein acetoxymethyl ester (BCECF-AM, fura-2 and [3H]GABA loaded synaptosomes, respectively. In the presence of Na+ at a physiological concentration (147 mM), nigericin (0.5 microM) elevates [Nai] from 20 to 50 mM, increases the pHi, 0.16 pH units, elevates four fold the [Cai] at expense of external Ca2+ and markedly increases (more than five fold) the release of [3H]GABA. In the absence of a Na+ concentration gradient (i.e. when the external Na+ concentration equals the [Nai]), the same concentration (0.5 microM) of nigericin causes the opposite effect on the pHi (acidifies the synaptosomal interior), does not modify the [Nai] and is practically unable to elevate the [Cai] or to increase [3H]GABA release. Only with higher concentrations of nigericin than 0.5 microM the ionophore is able to elevate the [Cai] and to increase the release of [3H]GABA under the conditions in which the net Na+ movements are eliminated. These results clearly show that under physiological conditions (147 mM external Na+) nigericin behaves as a Na+/H+ ionophore, and all its effects are triggered by the entrance of Na+ in exchange for H+ through the ionophore itself. Nigericin behaves as a K+/H+ ionophore in synaptosomes just when the net Na+ movements are eliminated (i.e. under conditions in which the external and the internal Na+ concentrations are equal). In summary care must be taken when using the putative K+/H+ ionophore nigericin as an experimental tool in synaptosomes, as under standard conditions (i.e. in the presence of high external Na+) nigericin behaves as a Na+/H+ ionophore.