Skip to main content
NCBI home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1993 Oct;470:575–600. doi: 10.1113/jphysiol.1993.sp019876

Calcium-mediated modulation of N-methyl-D-aspartate (NMDA) responses in cultured rat hippocampal neurones.

L Vyklický Jr 1
PMCID: PMC1143935  PMID: 8308745

Abstract

1. Agonist-independent (inactivation) and agonist-induced (desensitization) refractory states of N-methyl-D-aspartate (NMDA) receptors were studied on cultured rat hippocampal neurones using whole-cell and inside-out patch-clamp techniques and a fast perfusion system. 2. Shortly after whole-cell formation, application of 100 microM NMDA in the presence of 10 microM glycine and 0.2 mM [Ca2+]o induced membrane currents that desensitized by 23%. Repeated application of NMDA at 30 s intervals resulted in a progressive increase in the degree and rate of onset of NMDA receptor desensitization. 3. Test responses to NMDA recorded in the presence of 0.2 mM [Ca2+]o were reversibly inactivated by 60% following a train of ten 1 s applications of NMDA delivered at 0.5 Hz in the presence of 2 mM [Ca2+]o; similar results were obtained with 2 mM [Sr2+]o and 2 mM [Ba2+]o. In the presence of Ca2+ or Sr2+, desensitization during the train of responses to NMDA increased by 14 and 19% respectively, while with Ba2+ there was no increase in desensitization. 4. In the presence of 0.2 mM [Ca2+]o at a holding potential of -60 mV, or in the presence of 2 mM [Ca2+]o at a holding potential of +50 mV, a train of ten applications of NMDA failed to induce either inactivation or an increase in desensitization of test responses to NMDA. These results suggest an important role for [Ca2+]o in the induction of both inactivation and desensitization of NMDA receptors. 5. Increasing the intracellular calcium concentration, [Ca2+]i, via repeated activation of voltage-gated Ca2+ channels, resulted in a reversible inactivation of test responses to NMDA by 35% but failed to increase desensitization. In neurones dialysed with intracellular solution containing 2.5 mM Ca2+ NMDA receptor desensitization was similar to that in neurones dialysed with 10 nM Ca2+. 6. Block of NMDA receptor-channels by 2 mM [Mg2+]o during the train application of NMDA prevented the induction of both inactivation and desensitization. In contrast 3 mM [Mg2+]i was ineffective. 7. The magnitude of both inactivation and desensitization of NMDA receptors was not affected by intracellular dialysis of ATP, the non-hydrolysable ATP analogue 5'-adenylylimido-diphosphate (AMP-PNP), different Ca2+ chelators (EGTA or BAPTA), the Ca(2+)-activated protease inhibitor (leupeptin), dithiothreitol, or the phosphatase inhibitors (okadaic acid and a calcineurin inhibitor). 8. Application of 2.5 mM Ca2+ to the cytoplasmic side of inside-out patches induced inactivation of NMDA responses similar in magnitude to the inactivation seen in whole-cell recording.(ABSTRACT TRUNCATED AT 400 WORDS)

Full text

PDF
575

Selected References

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

  1. Aizenman E., Lipton S. A., Loring R. H. Selective modulation of NMDA responses by reduction and oxidation. Neuron. 1989 Mar;2(3):1257–1263. doi: 10.1016/0896-6273(89)90310-3. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bading H., Greenberg M. E. Stimulation of protein tyrosine phosphorylation by NMDA receptor activation. Science. 1991 Aug 23;253(5022):912–914. doi: 10.1126/science.1715095. [DOI] [PubMed] [Google Scholar]
  4. Benveniste M., Clements J., Vyklický L., Jr, Mayer M. L. A kinetic analysis of the modulation of N-methyl-D-aspartic acid receptors by glycine in mouse cultured hippocampal neurones. J Physiol. 1990 Sep;428:333–357. doi: 10.1113/jphysiol.1990.sp018215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brady R. J., Smith K. L., Swann J. W. Calcium modulation of the N-methyl-D-aspartate (NMDA) response and electrographic seizures in immature hippocampus. Neurosci Lett. 1991 Mar 11;124(1):92–96. doi: 10.1016/0304-3940(91)90829-i. [DOI] [PubMed] [Google Scholar]
  6. Chen L., Huang L. Y. Sustained potentiation of NMDA receptor-mediated glutamate responses through activation of protein kinase C by a mu opioid. Neuron. 1991 Aug;7(2):319–326. doi: 10.1016/0896-6273(91)90270-a. [DOI] [PubMed] [Google Scholar]
  7. Chizhmakov I. V., Kiskin N. I., Krishtal O. A. Two types of steady-state desensitization of N-methyl-D-aspartate receptor in isolated hippocampal neurones of rat. J Physiol. 1992 Mar;448:453–472. doi: 10.1113/jphysiol.1992.sp019051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clark G. D., Clifford D. B., Zorumski C. F. The effect of agonist concentration, membrane voltage and calcium on N-methyl-D-aspartate receptor desensitization. Neuroscience. 1990;39(3):787–797. doi: 10.1016/0306-4522(90)90261-2. [DOI] [PubMed] [Google Scholar]
  9. Cohen P., Holmes C. F., Tsukitani Y. Okadaic acid: a new probe for the study of cellular regulation. Trends Biochem Sci. 1990 Mar;15(3):98–102. doi: 10.1016/0968-0004(90)90192-e. [DOI] [PubMed] [Google Scholar]
  10. Croall D. E., DeMartino G. N. Calcium-activated neutral protease (calpain) system: structure, function, and regulation. Physiol Rev. 1991 Jul;71(3):813–847. doi: 10.1152/physrev.1991.71.3.813. [DOI] [PubMed] [Google Scholar]
  11. Halpain S., Greengard P. Activation of NMDA receptors induces rapid dephosphorylation of the cytoskeletal protein MAP2. Neuron. 1990 Sep;5(3):237–246. doi: 10.1016/0896-6273(90)90161-8. [DOI] [PubMed] [Google Scholar]
  12. Hashimoto Y., Perrino B. A., Soderling T. R. Identification of an autoinhibitory domain in calcineurin. J Biol Chem. 1990 Feb 5;265(4):1924–1927. [PubMed] [Google Scholar]
  13. Huganir R. L., Greengard P. Regulation of neurotransmitter receptor desensitization by protein phosphorylation. Neuron. 1990 Nov;5(5):555–567. doi: 10.1016/0896-6273(90)90211-w. [DOI] [PubMed] [Google Scholar]
  14. Johnson J. W., Ascher P. Voltage-dependent block by intracellular Mg2+ of N-methyl-D-aspartate-activated channels. Biophys J. 1990 May;57(5):1085–1090. doi: 10.1016/S0006-3495(90)82626-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. KATZ B., THESLEFF S. A study of the desensitization produced by acetylcholine at the motor end-plate. J Physiol. 1957 Aug 29;138(1):63–80. doi: 10.1113/jphysiol.1957.sp005838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Klee C. B., Draetta G. F., Hubbard M. J. Calcineurin. Adv Enzymol Relat Areas Mol Biol. 1988;61:149–200. doi: 10.1002/9780470123072.ch4. [DOI] [PubMed] [Google Scholar]
  17. Legendre P., Rosenmund C., Westbrook G. L. Inactivation of NMDA channels in cultured hippocampal neurons by intracellular calcium. J Neurosci. 1993 Feb;13(2):674–684. doi: 10.1523/JNEUROSCI.13-02-00674.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lerma J., Zukin R. S., Bennett M. V. Glycine decreases desensitization of N-methyl-D-aspartate (NMDA) receptors expressed in Xenopus oocytes and is required for NMDA responses. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2354–2358. doi: 10.1073/pnas.87.6.2354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Llano I., Leresche N., Marty A. Calcium entry increases the sensitivity of cerebellar Purkinje cells to applied GABA and decreases inhibitory synaptic currents. Neuron. 1991 Apr;6(4):565–574. doi: 10.1016/0896-6273(91)90059-9. [DOI] [PubMed] [Google Scholar]
  20. MacDermott A. B., Mayer M. L., Westbrook G. L., Smith S. J., Barker J. L. NMDA-receptor activation increases cytoplasmic calcium concentration in cultured spinal cord neurones. 1986 May 29-Jun 4Nature. 321(6069):519–522. doi: 10.1038/321519a0. [DOI] [PubMed] [Google Scholar]
  21. MacDonald J. F., Mody I., Salter M. W. Regulation of N-methyl-D-aspartate receptors revealed by intracellular dialysis of murine neurones in culture. J Physiol. 1989 Jul;414:17–34. doi: 10.1113/jphysiol.1989.sp017674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mayer M. L., MacDermott A. B., Westbrook G. L., Smith S. J., Barker J. L. Agonist- and voltage-gated calcium entry in cultured mouse spinal cord neurons under voltage clamp measured using arsenazo III. J Neurosci. 1987 Oct;7(10):3230–3244. doi: 10.1523/JNEUROSCI.07-10-03230.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mayer M. L., Vyklicky L., Jr, Clements J. Regulation of NMDA receptor desensitization in mouse hippocampal neurons by glycine. Nature. 1989 Mar 30;338(6214):425–427. doi: 10.1038/338425a0. [DOI] [PubMed] [Google Scholar]
  24. Mayer M. L., Vyklicky L., Jr, Westbrook G. L. Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones. J Physiol. 1989 Aug;415:329–350. doi: 10.1113/jphysiol.1989.sp017724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mayer M. L., Westbrook G. L. Permeation and block of N-methyl-D-aspartic acid receptor channels by divalent cations in mouse cultured central neurones. J Physiol. 1987 Dec;394:501–527. doi: 10.1113/jphysiol.1987.sp016883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mayer M. L., Westbrook G. L. The action of N-methyl-D-aspartic acid on mouse spinal neurones in culture. J Physiol. 1985 Apr;361:65–90. doi: 10.1113/jphysiol.1985.sp015633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ruppersberg J. P., Stocker M., Pongs O., Heinemann S. H., Frank R., Koenen M. Regulation of fast inactivation of cloned mammalian IK(A) channels by cysteine oxidation. Nature. 1991 Aug 22;352(6337):711–714. doi: 10.1038/352711a0. [DOI] [PubMed] [Google Scholar]
  28. Sather W., Dieudonné S., MacDonald J. F., Ascher P. Activation and desensitization of N-methyl-D-aspartate receptors in nucleated outside-out patches from mouse neurones. J Physiol. 1992 May;450:643–672. doi: 10.1113/jphysiol.1992.sp019148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sather W., Johnson J. W., Henderson G., Ascher P. Glycine-insensitive desensitization of NMDA responses in cultured mouse embryonic neurons. Neuron. 1990 May;4(5):725–731. doi: 10.1016/0896-6273(90)90198-o. [DOI] [PubMed] [Google Scholar]
  30. Seubert P., Larson J., Oliver M., Jung M. W., Baudry M., Lynch G. Stimulation of NMDA receptors induces proteolysis of spectrin in hippocampus. Brain Res. 1988 Sep 13;460(1):189–194. doi: 10.1016/0006-8993(88)91222-x. [DOI] [PubMed] [Google Scholar]
  31. Siman R., Noszek J. C. Excitatory amino acids activate calpain I and induce structural protein breakdown in vivo. Neuron. 1988 Jun;1(4):279–287. doi: 10.1016/0896-6273(88)90076-1. [DOI] [PubMed] [Google Scholar]
  32. Traynelis S. F., Cull-Candy S. G. Pharmacological properties and H+ sensitivity of excitatory amino acid receptor channels in rat cerebellar granule neurones. J Physiol. 1991 Feb;433:727–763. doi: 10.1113/jphysiol.1991.sp018453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  34. Vyklický L., Jr, Benveniste M., Mayer M. L. Modulation of N-methyl-D-aspartic acid receptor desensitization by glycine in mouse cultured hippocampal neurones. J Physiol. 1990 Sep;428:313–331. doi: 10.1113/jphysiol.1990.sp018214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Vyklický L., Jr, Vlachová V., Krůsek J. The effect of external pH changes on responses to excitatory amino acids in mouse hippocampal neurones. J Physiol. 1990 Nov;430:497–517. doi: 10.1113/jphysiol.1990.sp018304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wang K. K. Developing selective inhibitors of calpain. Trends Pharmacol Sci. 1990 Apr;11(4):139–142. doi: 10.1016/0165-6147(90)90060-L. [DOI] [PubMed] [Google Scholar]
  37. Zorumski C. F., Yang J., Fischbach G. D. Calcium-dependent, slow desensitization distinguishes different types of glutamate receptors. Cell Mol Neurobiol. 1989 Mar;9(1):95–104. doi: 10.1007/BF00711446. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES