Skip to main content
PMC home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1996 Jul 15;494(Pt 2):479–492. doi: 10.1113/jphysiol.1996.sp021507

Identification of a native low-conductance NMDA channel with reduced sensitivity to Mg2+ in rat central neurones.

A Momiyama 1, D Feldmeyer 1, S G Cull-Candy 1
PMCID: PMC1160649  PMID: 8842006

Abstract

1. We have identified a new type of NMDA channel in rat central neurones that express mRNA for the NR2D subunit. We have examined single NMDA channels in cerebellar Purkinje cells (which possess NR1 and 2D), deep cerebellar nuclei (NR1, 2A, 2B and 2D) and spinal cord dorsal horn neurones (NR1, 2B and 2D). 2. In Purkinje cells, NMDA opened channels with a main conductance of 37.9 +/- 1.1 pS and a subconductance of 17.8 +/- 0.7 pS, with frequent transitions between the two levels. 3. NMDA activated low-conductance ('38/18 pS') events (along with high-conductance--'50/40 pS'--openings) in some patches from deep cerebellar nuclei and dorsal horn neurones. Our evidence suggests that 38/18 pS and 50/40 pS events arose from distinct types of NMDA receptors. 4. The transitions for 38/18 pS events were asymmetrical: steps from 38 to 18 pS were more frequent (72.2%) than steps from 18 to 38 pS. This feature appeared common to the 38/18 pS events in all three cell types, suggesting similarity in the low-conductance channels. 5. The 38/18 pS channels in Purkinje cells exhibited characteristic NMDA receptor properties, including requirement for glycine, antagonism by D-2-amino-5-phosphonopentanoic acid (D-AP5) and 7-chlorokynurenic acid, and voltage-dependent block by extracellular Mg2+. 6. The mean open time for the 38 pS state (0.74 +/- 0.07 ms) was significantly briefer than that for the 18 pS state (1.27 +/- 0.18 ms). 7. Mg2+ block of low-conductance NMDA channels in Purkinje cells was less marked than block of 50/40 pS channels in cerebellar granule cells. 8. The time course of appearance of 38/18 pS NMDA channels matched the expression of mRNA for the NR2D subunit. Thus 38/18 pS events were present in > 70% of Purkinje cell patches in 0- to 8-day-old animals, and absent by postnatal day 12. 9. We propose that the 38/18 pS NMDA channels identified here (associated with the NR2D subunit), and the other low-conductance NMDA channel associated with the NR2C subunit, may together constitute a functionally distinct subclass of native NMDA receptors.

Full text

PDF
479

Images in this article

488Figure 8
on p.488

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akazawa C., Shigemoto R., Bessho Y., Nakanishi S., Mizuno N. Differential expression of five N-methyl-D-aspartate receptor subunit mRNAs in the cerebellum of developing and adult rats. J Comp Neurol. 1994 Sep 1;347(1):150–160. doi: 10.1002/cne.903470112. [DOI] [PubMed] [Google Scholar]
  2. Ascher P., Bregestovski P., Nowak L. N-methyl-D-aspartate-activated channels of mouse central neurones in magnesium-free solutions. J Physiol. 1988 May;399:207–226. doi: 10.1113/jphysiol.1988.sp017076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ascher P., Nowak L. The role of divalent cations in the N-methyl-D-aspartate responses of mouse central neurones in culture. J Physiol. 1988 May;399:247–266. doi: 10.1113/jphysiol.1988.sp017078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carmignoto G., Vicini S. Activity-dependent decrease in NMDA receptor responses during development of the visual cortex. Science. 1992 Nov 6;258(5084):1007–1011. doi: 10.1126/science.1279803. [DOI] [PubMed] [Google Scholar]
  5. Crepel F., Audinat E. Excitatory amino acid receptors of cerebellar Purkinje cells: development and plasticity. Prog Biophys Mol Biol. 1991;55(1):31–46. doi: 10.1016/0079-6107(91)90010-p. [DOI] [PubMed] [Google Scholar]
  6. Crepel F., Delhaye-Bouchaud N., Dupont J. L. Fate of the multiple innervation of cerebellar Purkinje cells by climbing fibers in immature control, x-irradiated and hypothyroid rats. Brain Res. 1981 Jan;227(1):59–71. doi: 10.1016/0165-3806(81)90094-8. [DOI] [PubMed] [Google Scholar]
  7. Cull-Candy S. G., Usowicz M. M. Multiple-conductance channels activated by excitatory amino acids in cerebellar neurons. Nature. 1987 Feb 5;325(6104):525–528. doi: 10.1038/325525a0. [DOI] [PubMed] [Google Scholar]
  8. Cull-Candy S. G., Usowicz M. M. On the multiple-conductance single channels activated by excitatory amino acids in large cerebellar neurones of the rat. J Physiol. 1989 Aug;415:555–582. doi: 10.1113/jphysiol.1989.sp017736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farrant M., Cull-Candy S. G. Excitatory amino acid receptor-channels in Purkinje cells in thin cerebellar slices. Proc Biol Sci. 1991 Jun 22;244(1311):179–184. doi: 10.1098/rspb.1991.0067. [DOI] [PubMed] [Google Scholar]
  10. Farrant M., Feldmeyer D., Takahashi T., Cull-Candy S. G. NMDA-receptor channel diversity in the developing cerebellum. Nature. 1994 Mar 24;368(6469):335–339. doi: 10.1038/368335a0. [DOI] [PubMed] [Google Scholar]
  11. Hestrin S. Developmental regulation of NMDA receptor-mediated synaptic currents at a central synapse. Nature. 1992 Jun 25;357(6380):686–689. doi: 10.1038/357686a0. [DOI] [PubMed] [Google Scholar]
  12. Hollmann M., Boulter J., Maron C., Beasley L., Sullivan J., Pecht G., Heinemann S. Zinc potentiates agonist-induced currents at certain splice variants of the NMDA receptor. Neuron. 1993 May;10(5):943–954. doi: 10.1016/0896-6273(93)90209-a. [DOI] [PubMed] [Google Scholar]
  13. Ikeda K., Nagasawa M., Mori H., Araki K., Sakimura K., Watanabe M., Inoue Y., Mishina M. Cloning and expression of the epsilon 4 subunit of the NMDA receptor channel. FEBS Lett. 1992 Nov 16;313(1):34–38. doi: 10.1016/0014-5793(92)81178-o. [DOI] [PubMed] [Google Scholar]
  14. Ishii T., Moriyoshi K., Sugihara H., Sakurada K., Kadotani H., Yokoi M., Akazawa C., Shigemoto R., Mizuno N., Masu M. Molecular characterization of the family of the N-methyl-D-aspartate receptor subunits. J Biol Chem. 1993 Feb 5;268(4):2836–2843. [PubMed] [Google Scholar]
  15. Jahr C. E., Stevens C. F. Glutamate activates multiple single channel conductances in hippocampal neurons. Nature. 1987 Feb 5;325(6104):522–525. doi: 10.1038/325522a0. [DOI] [PubMed] [Google Scholar]
  16. Kourie J. I., Laver D. R., Junankar P. R., Gage P. W., Dulhunty A. F. Characteristics of two types of chloride channel in sarcoplasmic reticulum vesicles from rabbit skeletal muscle. Biophys J. 1996 Jan;70(1):202–221. doi: 10.1016/S0006-3495(96)79564-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kutsuwada T., Kashiwabuchi N., Mori H., Sakimura K., Kushiya E., Araki K., Meguro H., Masaki H., Kumanishi T., Arakawa M. Molecular diversity of the NMDA receptor channel. Nature. 1992 Jul 2;358(6381):36–41. doi: 10.1038/358036a0. [DOI] [PubMed] [Google Scholar]
  18. Llano I., Marty A., Armstrong C. M., Konnerth A. Synaptic- and agonist-induced excitatory currents of Purkinje cells in rat cerebellar slices. J Physiol. 1991 Mar;434:183–213. doi: 10.1113/jphysiol.1991.sp018465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mariani J., Changeux J. P. Ontogenesis of olivocerebellar relationships. I. Studies by intracellular recordings of the multiple innervation of Purkinje cells by climbing fibers in the developing rat cerebellum. J Neurosci. 1981 Jul;1(7):696–702. doi: 10.1523/JNEUROSCI.01-07-00696.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mayer M. L., Westbrook G. L., Guthrie P. B. Voltage-dependent block by Mg2+ of NMDA responses in spinal cord neurones. Nature. 1984 May 17;309(5965):261–263. doi: 10.1038/309261a0. [DOI] [PubMed] [Google Scholar]
  21. McBain C. J., Mayer M. L. N-methyl-D-aspartic acid receptor structure and function. Physiol Rev. 1994 Jul;74(3):723–760. doi: 10.1152/physrev.1994.74.3.723. [DOI] [PubMed] [Google Scholar]
  22. Meguro H., Mori H., Araki K., Kushiya E., Kutsuwada T., Yamazaki M., Kumanishi T., Arakawa M., Sakimura K., Mishina M. Functional characterization of a heteromeric NMDA receptor channel expressed from cloned cDNAs. Nature. 1992 May 7;357(6373):70–74. doi: 10.1038/357070a0. [DOI] [PubMed] [Google Scholar]
  23. Monyer H., Burnashev N., Laurie D. J., Sakmann B., Seeburg P. H. Developmental and regional expression in the rat brain and functional properties of four NMDA receptors. Neuron. 1994 Mar;12(3):529–540. doi: 10.1016/0896-6273(94)90210-0. [DOI] [PubMed] [Google Scholar]
  24. Monyer H., Sprengel R., Schoepfer R., Herb A., Higuchi M., Lomeli H., Burnashev N., Sakmann B., Seeburg P. H. Heteromeric NMDA receptors: molecular and functional distinction of subtypes. Science. 1992 May 22;256(5060):1217–1221. doi: 10.1126/science.256.5060.1217. [DOI] [PubMed] [Google Scholar]
  25. Moriyoshi K., Masu M., Ishii T., Shigemoto R., Mizuno N., Nakanishi S. Molecular cloning and characterization of the rat NMDA receptor. Nature. 1991 Nov 7;354(6348):31–37. doi: 10.1038/354031a0. [DOI] [PubMed] [Google Scholar]
  26. Nowak L., Bregestovski P., Ascher P., Herbet A., Prochiantz A. Magnesium gates glutamate-activated channels in mouse central neurones. Nature. 1984 Feb 2;307(5950):462–465. doi: 10.1038/307462a0. [DOI] [PubMed] [Google Scholar]
  27. Perkel D. J., Hestrin S., Sah P., Nicoll R. A. Excitatory synaptic currents in Purkinje cells. Proc Biol Sci. 1990 Aug 22;241(1301):116–121. doi: 10.1098/rspb.1990.0074. [DOI] [PubMed] [Google Scholar]
  28. Petralia R. S., Wang Y. X., Wenthold R. J. The NMDA receptor subunits NR2A and NR2B show histological and ultrastructural localization patterns similar to those of NR1. J Neurosci. 1994 Oct;14(10):6102–6120. doi: 10.1523/JNEUROSCI.14-10-06102.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rabacchi S., Bailly Y., Delhaye-Bouchaud N., Mariani J. Involvement of the N-methyl D-aspartate (NMDA) receptor in synapse elimination during cerebellar development. Science. 1992 Jun 26;256(5065):1823–1825. doi: 10.1126/science.1352066. [DOI] [PubMed] [Google Scholar]
  30. Rosenmund C., Legendre P., Westbrook G. L. Expression of NMDA channels on cerebellar Purkinje cells acutely dissociated from newborn rats. J Neurophysiol. 1992 Nov;68(5):1901–1905. doi: 10.1152/jn.1992.68.5.1901. [DOI] [PubMed] [Google Scholar]
  31. Sheng M., Cummings J., Roldan L. A., Jan Y. N., Jan L. Y. Changing subunit composition of heteromeric NMDA receptors during development of rat cortex. Nature. 1994 Mar 10;368(6467):144–147. doi: 10.1038/368144a0. [DOI] [PubMed] [Google Scholar]
  32. Stern P., Béhé P., Schoepfer R., Colquhoun D. Single-channel conductances of NMDA receptors expressed from cloned cDNAs: comparison with native receptors. Proc Biol Sci. 1992 Dec 22;250(1329):271–277. doi: 10.1098/rspb.1992.0159. [DOI] [PubMed] [Google Scholar]
  33. Tsuzuki K., Mochizuki S., Iino M., Mori H., Mishina M., Ozawa S. Ion permeation properties of the cloned mouse epsilon 2/zeta 1 NMDA receptor channel. Brain Res Mol Brain Res. 1994 Oct;26(1-2):37–46. doi: 10.1016/0169-328x(94)90071-x. [DOI] [PubMed] [Google Scholar]
  34. Tölle T. R., Berthele A., Zieglgänsberger W., Seeburg P. H., Wisden W. The differential expression of 16 NMDA and non-NMDA receptor subunits in the rat spinal cord and in periaqueductal gray. J Neurosci. 1993 Dec;13(12):5009–5028. doi: 10.1523/JNEUROSCI.13-12-05009.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wafford K. A., Bain C. J., Le Bourdelles B., Whiting P. J., Kemp J. A. Preferential co-assembly of recombinant NMDA receptors composed of three different subunits. Neuroreport. 1993 Sep 30;4(12):1347–1349. doi: 10.1097/00001756-199309150-00015. [DOI] [PubMed] [Google Scholar]
  36. Watanabe M., Mishina M., Inoue Y. Distinct spatiotemporal distributions of the N-methyl-D-aspartate receptor channel subunit mRNAs in the mouse cervical cord. J Comp Neurol. 1994 Jul 8;345(2):314–319. doi: 10.1002/cne.903450212. [DOI] [PubMed] [Google Scholar]
  37. Yamazaki M., Mori H., Araki K., Mori K. J., Mishina M. Cloning, expression and modulation of a mouse NMDA receptor subunit. FEBS Lett. 1992 Mar 23;300(1):39–45. doi: 10.1016/0014-5793(92)80160-i. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES