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Patch Clamp Combined with Voltage/Concentration Clamp to Determine the Kinetics and Voltage Dependency of N-Methyl-d-aspartate (NMDA) Receptor Open Channel Blockers

  • Chris G. ParsonsEmail author
  • Kate E. Gilling
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1183)

Abstract

Electrophysiological techniques can be used to great effect to help determine the mechanism of action of a compound. However, many factors can compromise the resulting data and their analysis, such as the speed of solution exchange, expression of additional ion channel populations including other ligand-gated receptors and voltage-gated channels, compounds having multiple binding sites, and current desensitization and rundown. In this chapter, such problems and their solutions are discussed and illustrated using data from experiments involving the uncompetitive NMDA receptor antagonist memantine. Memantine differs from many other NMDA receptor channel blockers in that it is well tolerated and does not cause psychotomimetic effects at therapeutic doses. Various electrophysiological parameters of NMDA-induced current blockade by memantine have been proposed to be important in determining therapeutic tolerability; potency, onset and offset kinetics, and voltage dependency. These were all measured using whole cell patch clamp techniques using hippocampal neurons. Full results are shown here for memantine, and these are summarized and compared to those from similar experiments with other NMDA channel blockers. The interpretation of these results is discussed, as are theories concerning the tolerability of NMDA channel blockers, with the aim of illustrating how electrophysiological data can be used to form and support a physiological hypothesis.

Key words

NMDA Uncompetitive Concentration dependence Concentration clamp Voltage dependence Kinetics 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Pharmacology, Non-Clinical ScienceMerz Pharmaceuticals GmbHFrankfurt am MainGermany
  2. 2.Charité-UniversitätsmedizinBerlinGermany

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