Extracellular mild acidosis decreases the Ca2+ permeability of the human NMDA receptors

NMDA receptors (NMDARs) are glutamate-gated ion channels involved in excitatory synaptic transmission and in others physiological processes such as synaptic plasticity and development. The overload of Ca²⁺ ions through NMDARs, caused by an excessive activation of receptors, leads to excitotoxic neuronal cell death. For this reason, the reduction of Ca²⁺ flux through NMDARs has been a central focus in finding therapeutic strategies to prevent neuronal cell damage.Extracellular H⁺ are allosteric modulators of NMDARs. Starting from previous studies showing that extracellular mild acidosis reduces NMDA-evoked whole cell currents, we analyzed the effects of this condition on the NMDARs Ca²⁺ permeability, measured as “fractional calcium current” (P_f, i.e. the percentage of the total current carried by Ca²⁺ ions), of human NMDARs NR1/NR2A and NR1/NR2B transiently transfected in HeLa cells. Extracellular mild acidosis significantly reduces P_f of both human NR1/NR2A and NR1/NR2B NMDARs, also decreasing single channel conductance in outside out patches for NR1/NR2A receptor. Reduction of Ca²⁺ flux through NMDARs was also confirmed in cortical neurons in culture. A comparative analysis of both NMDA evoked Ca²⁺ transients and whole cell currents showed that extracellular H⁺ differentially modulate the permeation of Na⁺ and Ca²⁺ through NMDARs.Our data highlight the synergy of two distinct neuroprotective mechanisms during acidosis: Ca²⁺ entry through NMDARs is lowered due to the modulation of both open probability and Ca²⁺ permeability. Furthermore, this study provides the proof of concept that it is possible to reduce Ca²⁺ overload in neurons modulating the NMDAR Ca²⁺ permeability.