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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Permeation and block of N-methyl-D-aspartic acid receptor channels by divalent cations in mouse cultured central neurones.

1. Spinal cord and hippocampal neurones in cell culture were voltage clamped using the tight-seal, whole-cell recording technique. The concentration of sodium and a series of divalent cations in the extracellular media was varied to study permeation through excitatory amino acid receptor channels activated by the selective agonists N-methyl-D-aspartic acid (NMDA), kainic acid and quisqualic acid. 2. On raising the extracellular calcium concentration, with [Na+]o held constant at 105 mM, the reversal potential of responses to NMDA shifted in the depolarizing direction. This shift was adequately described by the extended constant-field equation over the range 0.3-50 mM-calcium. Using ionic activity coefficients we calculate a value of PCa/PNa = 10. 6. Under the same experimental conditions the reversal potential of responses to kainic and quisqualic acids was much less affected by raising the calcium concentration, such that PCa/PNa = 0.15. A depolarizing shift of the NMDA reversal potential was also recorded during application of 20 mM-barium, strontium or manganese, suggesting permeation of these ions. The permeability sequence was Ca2+ greater than Ba2+ greater than Sr2+ much greater than Mn2+. No depolarizing shift of the NMDA reversal potential occurred during application of 20 mM-cobalt, magnesium or nickel. 3. In experiments in which the extracellular Na+ concentration was varied the extended constant-field equation was adequate in predicting shifts of the NMDA reversal potential recorded on varying [Na+]o over the range 50-150 mM, but failed to accurately predict the reversal potential of responses to NMDA with 10 mM-[Ca2+]o and only 10 or 20 mM-[Na+]o. These results imply an apparent increase in PCa/PNa on lowering [Na+]o and may result from interaction of permeant ions within the channel. 4. Barium and to a lesser extent calcium, but not strontium (all 20 mM), reduced the slope conductance of responses to NMDA recorded within +/- 15 mV of the reversal potential; over this limited range of membrane potential the current-voltage relationship remained linear in the presence of each of these ions. In contrast manganese produced a strong, voltage-dependent block of responses to NMDA, similar to that produced by magnesium, such that even close to the reversal potential the NMDA current-voltage relationship was highly non-linear. Thus manganese both permeates and blocks the NMDA receptor channel. 5. Raising the extracellular calcium concentration, from 0.1 to 5 mM, had two effects on the conductance mechanism activated by NMDA.(ABSTRACT TRUNCATED AT 400 WORDS)[1]


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