Skip to main content

Changes in AMPA receptor-spliced variant expression and shift in AMPA receptor spontaneous desensitization pharmacology during cerebellar granule cell maturation in vitro

Journal of Molecular Neuroscience volume 11, pages 23–41 (1998)Cite this article

Abstract

Using appropriate internal standards, quantitative reverse transcripase-polymerase chain reaction (RT-PCR), and cerebellar granule cell (CG) in primary cultures we have quantified the expression of mRNAs encoding for GluR1–4 dl-α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor subunits during neuronal maturation in vitro. GluR1 is the mRNA that increases during CG maturation; the expression changes of the other GluR mRNAs are minimal and the translation products of these mRNAs change with a similar pattern. During CG maturation, there is an 8- to 10-fold increase in the GluR1 FLOP mRNA and a twofold increase in the expression of FLOP mRNA for GluR4 and GluR4C. The GluR1 FLIP mRNA increases, but by a smaller extent. We found that the GluR2 mRNA is completely edited at its Q/R site during CG maturation. The increase on the expression of GluR1 FLIP and FLOP and of GluR4 FLOP mRNA variants during development is associated with a 10-fold increase in AMPA-mediated Na+ currents and in the increased amplification of this current by 7-chloro-3-methyl-3,4-dihydro-2H-1,2,4benzothiadiazine S-S-dioxide (IDRA21) or by 6-chloro-3,4dihydro-3-(2-norbornen-5-yl)-7-sulfamoyl-1,2,4-benzothiadiazine 1,1 dioxide (cyclothiazide [CT]).

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  • Angus-Silver R., Traynelis S. F., and Cull-Candy S. G. (1992) Rapid-time-course miniature and evoked excitatory currents at cerebellar synapses in situ. Nature 355, 163–166.

    Article  Google Scholar 

  • Arai A., Guidotti A., Costa E., and Lynch G. (1996a) Effect of the AMPA receptor modulator IDRA21 on LTP in Hippocampal slices. Neuroreport 7, 2211–2215.

    PubMed  CAS  Article  Google Scholar 

  • Arai A., Kessler M., Rogers G., and Lynch G. (1996b) Effects of a memory-enhancing drug on DL-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor currents and synaptic transmission in hippocampus. J. Pharmacol. Exp. Ther. 278, 627–638.

    PubMed  CAS  Google Scholar 

  • Bennet J. A. and Dingledine R. (1995) Topology profile for a glutamate receptor: three transmembrane domains and a channel-lining reentrant membrane loop. Neuron 14, 373–384.

    Article  Google Scholar 

  • Bertolino M., Baraldi M., Parenti C., Braghiroli D., DiBella M., Vicini S., and Costa E. (1993) Modulation of AMPA/Kainate receptors by analogues of diazoxide and cyclothiazide in thin slices of rat hippocampus. Recepto. Channels 1, 267–278.

    CAS  Google Scholar 

  • Burnashev N., Monyer H., Seeburg P. H., and Sakmann B. (1992) Divalent ion permeability of AMPA receptor channels is dominated by the edited form of a single subunit. Neuron 8, 189–198.

    PubMed  Article  CAS  Google Scholar 

  • Chew L.-J., Fleck M. W., Wright P., Scherer S. E., Mayer M. L., and Gallo V. (1997) Growth factor-induced transcription of GluR1 increases functional AMPA receptor density in glial progenitor cells. J. Neurosci. 17, 227–240.

    PubMed  CAS  Google Scholar 

  • Chirgwin J. M., Przybyla A. E., MacDonald R. J., and Rutter W. J. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18, 5294–5299.

    PubMed  Article  CAS  Google Scholar 

  • Condorelli D. F., Dell’Albani P., Aronica E., Genazzani A. A., Casabona G., Corsaro M., Balazs R., and Nicoletti F. (1993) Growth conditions differentially regulate the expression of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor subunits in cultured neurons. J. Neurochem. 61, 2133–2139.

    PubMed  Article  CAS  Google Scholar 

  • Connolly C. N., Krishek B. J., McDonald B. J., Smart T. G., and Moss S. J. (1996) Assembly of cell surface expression of heteromeric and homomeric α-aminobutyric acid type A receptors. J. Biol. Chem. 271, 89–96.

    PubMed  Article  CAS  Google Scholar 

  • Copani A., Genazzani A. A., Aleppo G., Casabona G., Canonico P. L., Scapagnini U., and Nicoletti F. (1992) Nootropic drugs positively modulate α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-sensitive glutamate receptors in neuronal cultures. J. Neurochem. 58, 1199–1204.

    PubMed  Article  CAS  Google Scholar 

  • Donevan S. D. and Rogawski M. A. (1993) GYKI 52466, a 2,3 benzodiazepine, is a highly selective, noncompetitive antagonist of AMPA/Kainate receptor responses. Neuron 10, 51–59.

    PubMed  Article  CAS  Google Scholar 

  • Fleck M. W., Bahring R., Patneau D. K., and Mayer M. L. (1996) AMPA receptor heterogeneity in rat hippocampal neurons revealed by differential sensitivity to cyclothiazide. J. Neurophysiol. 75, 2322–2333.

    PubMed  CAS  Google Scholar 

  • Forti L. and Pietrobon D. (1993) Functional diversity of L-type calcium channels in rat cerebellar neurons Neuron 10, 437–450.

    PubMed  Article  CAS  Google Scholar 

  • Gallo V., Upson L. M., Hayes W. P., Vyklicky L. Jr., Winters C., and Buonanno A. (1992) Molecular cloning and developmental analysis of a new glutamate receptor subunit isoform in cerebellum. J. Neurosci. 12, 1010–1023.

    PubMed  CAS  Google Scholar 

  • Geiger J. R., Melcher T., Koh D. S., Sakmann B., Seeburg P. H., Jonas P., and Monyer H. (1995) Relative abundance of subunit mRNAs determines gating and Ca2+ permeability of AMPA receptors in principal neurons and interneurons in rat CNS. Neuron 15, 193–204.

    PubMed  Article  CAS  Google Scholar 

  • Gouliaev A. H. and Senning A. (1994) Piracetam and other structurally related nootropics. Brain Res. 19, 180–222.

    Article  CAS  Google Scholar 

  • Grayson D. R., Bovolin P., and Santi M. R. (1993) Absolute quantitation of γ-aminobutyric acid: a receptor subunit messenger RNAs by competitive polymerase chain reaction. Methods Neurosci. 12, 191–208.

    CAS  Google Scholar 

  • Grynkiewicz G., Poenie M., and Tsien R. Y. (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260, 3440–3450.

    PubMed  CAS  Google Scholar 

  • Hack N. J., Sluiter A. A., and Balazs R. (1995) AMPA receptors in cerebellar granule cells during development in culture. Brain Res. Dev. Brain Res. 87, 55–61.

    PubMed  Article  CAS  Google Scholar 

  • Hamill O. P., Marty A., Neher E., Sakmann B., and Sigworth F. J. (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch. 391, 85–100.

    PubMed  Article  CAS  Google Scholar 

  • Harootunian A. T., Kao J. P., Eckert B. K., and Tsien R. Y. (1991) Fluorescence ratio imaging of cytosolic free Na+ in individual fibroblasts and lymphocytes. J. Biol. Chem. 264, 19,458–19,467.

    Google Scholar 

  • Hollmann M. and Heinemann S. (1994) Cloned glutamate receptors. Annu. Rev. Neurosci. 17, 31–108.

    PubMed  Article  CAS  Google Scholar 

  • Hollmann M., Maron C., and Heinemann S. (1994) N-glycosylation site tagging suggests a three transmembrane domain topology for the glutamate receptor GluR1. Neuron 13, 1331–1343.

    PubMed  Article  CAS  Google Scholar 

  • Impagnatiello F., Oberto A., Longone P., Costa E., and Guidotti A. (1997) 7-Chloro-3-methyl-3,4-dihydro-2H-1,2,4-benzothiadiazine-S,S-dioxide: a partial modulator of AMPA receptor desensitization devoid of neurotoxicity. Proc. Natl. Acad. Sci. USA 94, 7053–7058.

    PubMed  Article  CAS  Google Scholar 

  • Isaacson J. S. and Nicoll R. A. (1994) Aniracetam reduces glutamate receptor desensitization and slows the decay of fast excitatory synaptic currents in the hippocampus. Proc. Natl. Acad. Sci. USA 88, 10,936–10,940.

    Google Scholar 

  • Ito I., Tanabe S., Kohda A., and Sugiyama H. (1990) Allosteric potentiation of quisqualate receptors by a nootropic drug aniracetam. J. Physiol. Lond. 424, 533–543.

    PubMed  CAS  Google Scholar 

  • Johansen T. H., Chaudhary A., and Verdoorn T. A. (1995) Interaction among GYKI-52466, cyclothiazide, and aniracetam at recombinant AMPA and kainate receptors. Mol. Pharmacol. 48, 946–955.

    PubMed  CAS  Google Scholar 

  • Kiedrowski L. and Costa E. (1995) Glutamate-induced destabilization of intracellular calcium concentration homeostasis in cultured cerebellar granule cells: role of mitochondria in calcium buffering. Mol. Pharmacol. 47, 140–147.

    PubMed  CAS  Google Scholar 

  • Laemlli U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.

    Article  Google Scholar 

  • Monyer H., Seeburg P. H., and Wisden W. (1991) Glutamate-operated channels: developmentally early and mature forms arise by alternative splicing. Neuron 6, 799–810.

    PubMed  Article  CAS  Google Scholar 

  • Mosbacher J., Schoepfer R., Monyer H., Burnashev N., Seeburg P. H., and Ruppersberg J. P. (1994) A molecular determinant for submillisecond desensitization in glutamate receptors Science 266, 1059–1062.

    PubMed  Article  CAS  Google Scholar 

  • Moudy A. M., Yamada K. A., and Rothman S. M. (1994) Rapid desensitization determines the pharmacology of glutamate neurotoxicity Neuropharmacol. 33, 953–962.

    Article  CAS  Google Scholar 

  • Nakanishi N., Shneider N. A., and Axel R. (1990) A family of glutamate receptors genes: evidence for the formation of heteromultimeric receptors with distinct channel properties. Neuron 5, 569–581.

    PubMed  Article  CAS  Google Scholar 

  • Nicoletti F., Wroblewski J. T., Novelli A., Alho H., Guidotti A., and Costa E. (1986) The activation of inositol phospholipid metabolism as a signal-transducing system for excitatory amino acids in primary cultures of cerebellar granule cells J. Neurosci. 6, 1905–1911.

    PubMed  CAS  Google Scholar 

  • Partin K. M., Fleck M. W., and Mayer M. L. (1996) AMPA receptor FLIP/FLOP Mutants affecting deactivation, desensitization and modulation by cyclothiazide, aniracetam and thiocyanate. J. Neurosci. 16, 6634–6647.

    PubMed  CAS  Google Scholar 

  • Partin K. M., Patneau D. K., and Mayer M. L. (1994) Cyclothiazide differentially modulates desensitization of α-amino-3-hydroxy-5-methil-4-isoxazole-propionic acid receptor splice variants. Mol. Pharmacol. 46, 129–138.

    PubMed  CAS  Google Scholar 

  • Pollard H., Heron A., Moreau J., Ben-Ari Y., and Khrestchatisky M. (1993) Alterations of the GluR-B AMPA receptor FLIP/FLOP expression in kainate-induced epilepsy and ischemia. Neuroscience 57, 545–554.

    PubMed  Article  CAS  Google Scholar 

  • Rheinalt Parri H. and Lansman J. B. (1996) Multiple components of Ca2+ channels facilitation in cerebellar granule cells: expression of facilitation during development in culture. J. Neurosci. 16, 4890–4902.

    Google Scholar 

  • Sambrook J., Fritsch E. F., and Maniatis T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  • Seeburg P. H. (1993) The TIPS/TINS lecture: the molecular biology of mammalian glutamate receptor channels. Trends Pharmacol. Sci. 14, 297–303.

    PubMed  Article  CAS  Google Scholar 

  • Sekiguchi M., Fleck M. W., Mayer M., Takeo J., Chiba Y., Yamashita S., and Wada K. (1997) A novel allosteric potentiator of AMPA receptors: 4-[2-(Phenylsulfonylamino) ethylthio]-2, 6-difluorophenoxyacetamide. J. Neurosci. 17, 5760–5771.

    PubMed  CAS  Google Scholar 

  • Sommer B., Keinanen K., Verdoorn T. A., Wisden W., Burnashev N., Herb A., Kohler M., Takagi T., Sakmann B., and Seeburg P. H. (1990) FLIP and FLOP: a cell-specific functional switch in glutamate-operated channels of the CNS. Science 249, 1580–1585.

    PubMed  Article  CAS  Google Scholar 

  • Staubli U., Perez Y., Xu F. B., Rogers G., Ingvar M., Stone-Elander S., and Lynch G. (1994) Centrally active modulators of glutamate receptors facilitate the induction of long-term potentiation in vivo. Proc. Natl. Acad. Sci. USA 91, 11,158–11,162.

    CAS  Google Scholar 

  • Stern-Bach Y., Bettler B., Hartley M., Sheppard P. O., O’Hara P. J., and Heinemann, S. F. (1994) Agonist selectivity of glutamate receptors is specified by two domains structurally related to bacterial amino binding proteins. Neuron 13, 1345–1357.

    PubMed  Article  CAS  Google Scholar 

  • Swanson G. T., Kamboj S. K., and Cull-Candy S. G. (1997) Single-channel properties of recombinant AMPA receptors depend on RNA editing, splice variation, and subunit composition. J. Neurosci. 17, 58–69.

    PubMed  CAS  Google Scholar 

  • Thompson D. M., Guidotti A., Di Bella M., and Costa E. (1995) 7-Chloro-3-Methyl-3,4-dihydro-2H-1,2,4-benzothiadiazineS,S-Dioxide (IDRA21): a congener of aniracetam, potently abates pharmacologically induced cognition impairments in patas monkeys. Proc. Natl. Acad. Sci. USA 92, 7667–7671.

    PubMed  Article  CAS  Google Scholar 

  • Wenthold R. J., Yokotani N., Doi K., and Wada K. (1992) Immunochemical characterization of the non-NMDA glutamate receptor using subunit-specific antibodies. J. Biol. Chem. 267, 501–507.

    PubMed  CAS  Google Scholar 

  • Wyllie D. J., Traynelis S. F., and Cull-Candy S. G. (1993) Evidence for more than one type of non-NMDA receptor in outside-out patches from cerebellar granule cells of the rat. J. Physiol. 463, 193–226.

    PubMed  CAS  Google Scholar 

  • Yamada K. A. and Rothman S. M. (1992) Diazoxide blocks glutamate desensitization and prolongs excitatory postsynaptic currents in rat hippocampal neurons. J. Physiol. Lond. 458, 409–423.

    PubMed  CAS  Google Scholar 

  • Yamada K. A. and Tang C. M. (1993) Benzothiadiazines inhibit rapid glutamate receptor desensitization and enhances glutamatergic synaptic currents. J. Neurosci. 13, 3904–3915.

    PubMed  CAS  Google Scholar 

  • Zivkovic I., Thompson D. M., Bertolino M., Uzunov D., Di Bella M., Costa E., and Guidotti A. (1995) 7-Chloro-3-methyl-3-4-dihydro-2H-1,2,4benzothiadiazineS,S-dioxide (IDRA21): a benzothiadiazide derivative that enhance cognition by attenuating DL-α-amino-2,3-dihydro-5-methyl-3-oxo-4-isoxazolepropanoic acid (AMPA) receptor desensitization. J. Pharmacol. Exp. Ther. 272, 300–309.

    PubMed  CAS  Google Scholar 

Download references

Author information

Affiliations

  1. The Psychiatric Institute, Department of Psychiatry, University of Illinois College of Medicine, 60612, Chicago, IL

    Patrizia Longone, Francesco Impagnatiello, Jean-Marc Mienville, Erminio Costa & Alessandro Guidotti

Authors
  1. Patrizia Longone
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Francesco Impagnatiello
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jean-Marc Mienville
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Erminio Costa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alessandro Guidotti
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Drs. Longone and Impagnatiello contributed equally to this work and should be considered co-lead authors.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Longone, P., Impagnatiello, F., Mienville, JM. et al. Changes in AMPA receptor-spliced variant expression and shift in AMPA receptor spontaneous desensitization pharmacology during cerebellar granule cell maturation in vitro. J Mol Neurosci 11, 23–41 (1998). https://doi.org/10.1385/JMN:11:1:23

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1385/JMN:11:1:23

Index Entries

  • AMPA receptors
  • cerebellar granule cells
  • cyclothiazide
  • IDRA21
  • glutamate receptors