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J Neurophysiol 79: 555-566, 1998;
0022-3077/98 $5.00
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The Journal of Neurophysiology Vol. 79 No. 2 February 1998, pp. 555-566
Copyright ©1998 The American Physiological Society

Functional and Pharmacological Differences Between Recombinant N-Methyl-D-Aspartate Receptors

Stefano Vicini1, Jian Feng Wang1, Jin Hong Li1, Wei Jian Zhu1, Yue Hua Wang2, JIAN HONG Luo2, Barry B. Wolfe2, and Dennis R. Grayson3

1 Department of Physiology and Biophysics and 2 Department of Pharmacology, Georgetown University School of Medicine, Washington, DC 20007; and 3 Departments of Psychiatry and Anatomy and Neurobiology, Neurosciences Research Center, Allegheny Singer Research Institute, Allegheny University of the Health Sciences, Pittsburgh, Pennsylvania 15212

Vicini, Stefano, Jian Feng Wang, Jin Hong Li, Wei Jian Zhu, Yue Hua Wang, Jian Hong Luo, Barry B. Wolfe, and Dennis R. Grayson. Functional and pharmacological differences between recombinant N-methyl-D-aspartate receptors. J. Neurophysiol. 79: 555-566, 1998. N-methyl-D-aspartic acid (NMDA) receptors transiently transfected into mammalian HEK-293 cells were characterized with subunit-specific antibodies and electrophysiological recordings. Deactivation time course recorded in response to fastL-glutamate pulses were studied in isolated and lifted cells, as well as in outside-out membrane patches excised from cells expressing recombinant NR1 subunits in combination with the NR2A, NR2B, NR2C, or NR2D NMDA receptor subunits. Transfected cells were preidentified by the fluorescence emitted from the coexpressed Aequorea victoria jellyfish Green Lantern protein. Currents generated by NR1/NR2A channels displayed double exponential deactivation time course being faster than that in NR1/NR2B or NR1/NR2C channels. However, a large decay variability was observed within each cotransfection, suggesting that mechanisms additional to subunit composition may also regulate deactivation time course. NR1/NR2D channels displayed slowly deactivating currents. Channel deactivation was fast and comparable among receptors obtained by cotransfecting five distinct spliced variants of the NR1 subunit, each with the NR2A subunit. Additionally, recovery from desensitization was slower for NR1/NR2B than for NR1/NR2A channels. The average deactivation time course of responses to brief L-glutamate applications in cells where NR1/NR2A/NR2B cDNAs were cotransfected at variable ratio was intermediate between those of the NR1/NR2A and NR1/NR2B channels. Although immunocytochemical evidence indicates that the majority of cells are cotransfected by all plasmids in triple transfection, our experimental condition did not allow for a tight control of the expression of NMDA receptor subunits. This produced the result that many cells were characterized by deactivation time course and haloperidol sensitivities of separate NR1/NR2A and NR1/NR2B subunit heteromers. We also speculate on the possible formation of channels resulting from the coassembly in the same receptor of NR1/NR2A/NR2B subunits from a minority of cells that gave responses to brief application of L-glutamate characterized by slow deactivation time course and decreased haloperidol sensitivity.




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S. S. Kumar and J. R. Huguenard
Pathway-Specific Differences in Subunit Composition of Synaptic NMDA Receptors on Pyramidal Neurons in Neocortex
J. Neurosci., November 5, 2003; 23(31): 10074 - 10083.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
N. O. Dalby and I. Mody
Activation of NMDA Receptors in Rat Dentate Gyrus Granule Cells by Spontaneous and Evoked Transmitter Release
J Neurophysiol, August 1, 2003; 90(2): 786 - 797.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
M. Lopez de Armentia and P. Sah
Development and Subunit Composition of Synaptic NMDA Receptors in the Amygdala: NR2B Synapses in the Adult Central Amygdala
J. Neurosci., July 30, 2003; 23(17): 6876 - 6883.
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 J. Neurosci.Home page
Y. Yoshimura, T. Ohmura, and Y. Komatsu
Two Forms of Synaptic Plasticity with Distinct Dependence on Age, Experience, and NMDA Receptor Subtype in Rat Visual Cortex
J. Neurosci., July 23, 2003; 23(16): 6557 - 6566.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
S. G. Brickley, C. Misra, M. H. S. Mok, M. Mishina, and S. G. Cull-Candy
NR2B and NR2D Subunits Coassemble in Cerebellar Golgi Cells to Form a Distinct NMDA Receptor Subtype Restricted to Extrasynaptic Sites
J. Neurosci., June 15, 2003; 23(12): 4958 - 4966.
[Abstract] [Full Text] [PDF]

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 ScienceHome page
R. Kawakami, Y. Shinohara, Y. Kato, H. Sugiyama, R. Shigemoto, and I. Ito
Asymmetrical Allocation of NMDA Receptor {epsilon}2 Subunits in Hippocampal Circuitry
Science, May 9, 2003; 300(5621): 990 - 994.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
M. Martina, N. V Krasteniakov, and R. Bergeron
D-Serine differently modulates NMDA receptor function in rat CA1 hippocampal pyramidal cells and interneurons
J. Physiol., April 15, 2003; 548(2): 411 - 423.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
G. Losi, K. Prybylowski, Z. Fu, J. Luo, R. J Wenthold, and S. Vicini
PSD-95 regulates NMDA receptors in developing cerebellar granule neurons of the rat
J. Physiol., April 1, 2003; 548(1): 21 - 29.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
M. Townsend, A. Yoshii, M. Mishina, and M. Constantine-Paton
Developmental loss of miniature N-methyl-D-aspartate receptor currents in NR2A knockout mice
PNAS, February 4, 2003; 100(3): 1340 - 1345.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
E. Harvey-Girard and R. J. Dunn
Excitatory Amino Acid Receptors of the Electrosensory System: The NR1/NR2B N-Methyl-D-Aspartate Receptor
J Neurophysiol, February 1, 2003; 89(2): 822 - 832.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
L. Chen, M. Muhlhauser, and C. R. Yang
Glycine Tranporter-1 Blockade Potentiates NMDA-Mediated Responses in Rat Prefrontal Cortical Neurons In Vitro and In Vivo
J Neurophysiol, February 1, 2003; 89(2): 691 - 703.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
L. J Gentet and D. Ulrich
Strong, reliable and precise synaptic connections between thalamic relay cells and neurones of the nucleus reticularis in juvenile rats
J. Physiol., February 1, 2003; 546(3): 801 - 811.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
H. Ren, Y. Honse, B. J. Karp, R. H. Lipsky, and R. W. Peoples
A Site in the Fourth Membrane-associated Domain of the N-Methyl-D-aspartate Receptor Regulates Desensitization and Ion Channel Gating
J. Biol. Chem., January 3, 2003; 278(1): 276 - 283.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
D. E. Chapman, K. A. Keefe, and K. S. Wilcox
Evidence for Functionally Distinct Synaptic NMDA Receptors in Ventromedial Versus Dorsolateral Striatum
J Neurophysiol, January 1, 2003; 89(1): 69 - 80.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
K. Prybylowski, Z. Fu, G. Losi, L. M. Hawkins, J. Luo, K. Chang, R. J. Wenthold, and S. Vicini
Relationship between Availability of NMDA Receptor Subunits and Their Expression at the Synapse
J. Neurosci., October 15, 2002; 22(20): 8902 - 8910.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
B. H. Miller and A. C. Gore
N-Methyl-D-Aspartate Receptor Subunit Expression in GnRH Neurons Changes during Reproductive Senescence in the Female Rat
Endocrinology, September 1, 2002; 143(9): 3568 - 3574.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
J. C Pina-Crespo and A. J Gibb
Subtypes of NMDA receptors in new-born rat hippocampal granule cells
J. Physiol., May 15, 2002; 541(1): 41 - 64.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
J. Nabekura, T. Ueno, S. Katsurabayashi, A. Furuta, N. Akaike, and M. Okada
Reduced NR2A expression and prolonged decay of NMDA receptor-mediated synaptic current in rat vagal motoneurons following axotomy
J. Physiol., March 15, 2002; 539(3): 735 - 741.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
G. Losi, K. Prybylowski, Z. Fu, J. H. Luo, and S. Vicini
Silent Synapses in Developing Cerebellar Granule Neurons
J Neurophysiol, March 1, 2002; 87(3): 1263 - 1270.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
G. Woodhall, D. I. Evans, M. O. Cunningham, and R. S. G. Jones
NR2B-Containing NMDA Autoreceptors at Synapses on Entorhinal Cortical Neurons
J Neurophysiol, October 1, 2001; 86(4): 1644 - 1651.
[Abstract] [Full Text] [PDF]

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 Mol. Pharmacol.Home page
N. Chen, J. Ren, L. A. Raymond, and T. H. Murphy
Changes in Agonist Concentration Dependence That Are a Function of Duration of Exposure Suggest N-Methyl-D-aspartate Receptor Nonsaturation during Synaptic Stimulation
Mol. Pharmacol., February 1, 2001; 59(2): 212 - 219.
[Abstract] [Full Text]

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 J. Neurosci.Home page
L. Pasti, M. Zonta, T. Pozzan, S. Vicini, and G. Carmignoto
Cytosolic Calcium Oscillations in Astrocytes May Regulate Exocytotic Release of Glutamate
J. Neurosci., January 15, 2001; 21(2): 477 - 484.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
D. Anchisi, B. Scelfo, and F. Tempia
Postsynaptic Currents in Deep Cerebellar Nuclei
J Neurophysiol, January 1, 2001; 85(1): 323 - 331.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
J. D. Sinor, S. Du, S. Venneti, R. C. Blitzblau, D. N. Leszkiewicz, P. A. Rosenberg, and E. Aizenman
NMDA and Glutamate Evoke Excitotoxicity at Distinct Cellular Locations in Rat Cortical Neurons In Vitro
J. Neurosci., December 1, 2000; 20(23): 8831 - 8837.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
L. Cathala, C. Misra, and S. Cull-Candy
Developmental Profile of the Changing Properties of NMDA Receptors at Cerebellar Mossy Fiber-Granule Cell Synapses
J. Neurosci., August 15, 2000; 20(16): 5899 - 5905.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
C. Misra, S. G Brickley, D. J A Wyllie, and S. G Cull-Candy
Slow deactivation kinetics of NMDA receptors containing NR1 and NR2D subunits in rat cerebellar Purkinje cells
J. Physiol., June 1, 2000; 525(2): 299 - 305.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
C. Misra, S. G Brickley, M. Farrant, and S. G Cull-Candy
Identification of subunits contributing to synaptic and extrasynaptic NMDA receptors in Golgi cells of the rat cerebellum
J. Physiol., April 1, 2000; 524(1): 147 - 162.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
A. Momiyama
Distinct synaptic and extrasynaptic NMDA receptors identified in dorsal horn neurones of the adult rat spinal cord
J. Physiol., March 15, 2000; 523(3): 621 - 628.
[Abstract] [Full Text] [PDF]

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 J. Neurophysiol.Home page
G. Rumbaugh, K. Prybylowski, J. F. Wang, and S. Vicini
Exon 5 and Spermine Regulate Deactivation of NMDA Receptor Subtypes
J Neurophysiol, March 1, 2000; 83(3): 1300 - 1306.
[Abstract] [Full Text] [PDF]

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 J. Pharmacol. Exp. Ther.Home page
Y. Li-Smerin, E. Aizenman, and J. W. Johnson
Inhibition by Intracellular Mg2+ of Recombinant N-Methyl-D-aspartate Receptors Expressed in Chinese Hamster Ovary Cells
J. Pharmacol. Exp. Ther., March 1, 2000; 292(3): 1104 - 1110.
[Abstract] [Full Text]

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 J. Neurophysiol.Home page
K. R. Tovar, K. Sprouffske, and G. L. Westbrook
Fast NMDA Receptor-Mediated Synaptic Currents in Neurons From Mice Lacking the epsilon 2 (NR2B) Subunit
J Neurophysiol, January 1, 2000; 83(1): 616 - 620.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
G. Rumbaugh and S. Vicini
Distinct Synaptic and Extrasynaptic NMDA Receptors in Developing Cerebellar Granule Neurons
J. Neurosci., December 15, 1999; 19(24): 10603 - 10610.
[Abstract] [Full Text] [PDF]


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