Abstract
N-methyl-d-aspartate receptors (NMDARs) are ionotropic glutamate receptors that are essential for synaptic plasticity, learning and memory. Dysfunction of NMDARs has been implicated in many nervous system disorders; therefore, pharmacological modulation of NMDAR activity has great therapeutic potential. However, given the broad physiological importance of NMDARs, modulating their activity often has detrimental side effects precluding pharmaceutical use of many NMDAR modulators. One approach to possibly improve the therapeutic potential of NMDAR modulators is to identify compounds that modulate subsets of NMDARs. An obvious target for modulating NMDAR subsets is the many NMDAR subtypes produced through different combinations of NMDAR subunits. With seven identified genes that encode NMDAR subunits, there are many neuronal NMDAR subtypes with distinct properties and potentially differential pharmacological sensitivities. Study of NMDAR subtype-specific pharmacology is complicated in neurons, however, because most neurons express at least three NMDAR subtypes. Thus, use of an approach that permits study in isolation of a single receptor subtype is preferred. Additionally, the effects of drugs on agonist-activated responses typically depend on duration of agonist exposure. To evaluate drug effects on synaptic transmission, an approach should be used that allows for activation of receptor responses as brief as those observed during synaptic transmission, both in the absence and presence of drug. To address these issues, we designed a fast perfusion system capable of (1) delivering brief (~5 ms) and consistent applications of glutamate to recombinant NMDARs of known subunit composition, and (2) easily and quickly (~5 s) changing between glutamate applications in the absence and presence of drug.
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References
Traynelis SF, Wollmuth LP, McBain CJ et al (2010) Glutamate receptor ion channels: structure, regulation, and function. Pharmacol Rev 62:405–496
Barnham KJ, Masters CL, Bush AI (2004) Neurodegenerative diseases and oxidative stress. Nat Rev Drug Discov 3:205–214
Lau A, Tymianski M (2010) Glutamate receptors, neurotoxicity and neurodegeneration. Europ J Physiol 460:525–542
Duman RS, Aghajanian GK (2012) Synaptic dysfunction in depression: potential therapeutic targets. Science 338:68–72
Autry AE, Adachi M, Nosyreva E et al (2011) NMDA receptor blockade at rest triggers rapid behavioural antidepressant responses. Nature 475:91–95
Pittenger C, Sanacora G, Krystal JH (2007) The NMDA receptor as a therapeutic target in major depressive disorder. CNS Neurol Disord Drug Targets 6:101–115
Collins S, Sigtermans MJ, Dahan A et al (2010) NMDA receptor antagonists for the treatment of neuropathic pain. Pain Med 11:1726–1742
Palmer GC (2001) Neuroprotection by NMDA receptor antagonists in a variety of neuropathologies. Curr Drug Targets 2:241–271
Lipton SA (2004) Failures and successes of NMDA receptor antagonists: molecular basis for the use of open-channel blockers like memantine in the treatment of acute and chronic neurologic insults. NeuroRx 1:101–110
Paoletti P, Bellone C, Zhou Q (2013) NMDA receptor subunit diversity: impact on receptor properties, synaptic plasticity and disease. Nat Rev Neurosci 14:383–400
Lipton SA (2006) Paradigm shift in neuroprotection by NMDA receptor blockade: memantine and beyond. Nat Rev Drug Discov 5:160–170
Gladding CM, Raymond LA (2011) Mechanisms underlying NMDA receptor synaptic/extrasynaptic distribution and function. Mol Cell Neurosci 48:308–320
Witt A, Macdonald N, Kirkpatrick P (2004) Memantine hydrochloride. Nat Rev Drug Discov 3:109–110
Prommer EE (2012) Ketamine for pain: an update of uses in palliative care. J Palliat Med 15:474–483
Anitha M, Nandhu MS, Anju TR et al (2011) Targeting glutamate mediated excitotoxicity in Huntington’s disease: neural progenitors and partial glutamate antagonist–memantine. Med Hypotheses 76:138–140
Kotermanski SE, Wood JT, Johnson JW (2009) Memantine binding to a superficial site on NMDA receptors contributes to partial trapping. J Physiol 587:4589–4604
Kotermanski SE, Johnson JW (2009) Mg2+ imparts NMDA receptor subtype selectivity to the Alzheimer’s drug memantine. J Neurosci 29:2774–2779
Phelan MC (2006) Techniques for mammalian cell tissue culture. Curr Protoc Mol Biol Appendix 3: Appendix 3F
Hamill OP, Marty A, Neher E et al (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch 391:85–100
Vicini S, Wang JF, Li JH et al (1998) Functional and pharmacological differences between recombinant N-methyl-D-aspartate receptors. J Neurophysiol 79:555–566
Erreger K, Dravid SM, Banke TG et al (2005) Subunit-specific gating controls rat NR1/NR2A and NR1/NR2B NMDA channel kinetics and synaptic signalling profiles. J Physiol 563:345–358
Cull-Candy SG, Leszkiewicz DN (2004) Role of distinct NMDA receptor subtypes at central synapses. Sci STKE 2004(255):16
Tovar KR, McGinley MJ, Westbrook GL (2013) Triheteromeric NMDA receptors at hippocampal synapses. J Neurosci 33:9150–9160
Acknowledgements
The authors would like to thank Christen Shiber for excellent technical assistance and critically reading the manuscript. The authors would also like to thank Jim Buhrman for excellent technical assistance and helpful discussions regarding fast perfusion system design.
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Glasgow, N.G., Johnson, J.W. (2014). Whole-Cell Patch-Clamp Analysis of Recombinant NMDA Receptor Pharmacology Using Brief Glutamate Applications. In: Martina, M., Taverna, S. (eds) Patch-Clamp Methods and Protocols. Methods in Molecular Biology, vol 1183. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1096-0_2
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DOI: https://doi.org/10.1007/978-1-4939-1096-0_2
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