Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Functional Expression of GABAA Chloride Channels and Benzodiazepine Binding Sites in Baculovirus Infected Insect Cells

Bio/Technology volume 10, pages 679–681 (1992)Cite this article

Abstract

We have employed the baculovirus expression system for the production of insect cell membranes having GABA/benzo–diazepine binding sites. Three recombinant baculoviruses each harboring a different GABAA receptor cDNA were introduced into insect cells by simultaneous infection. Infected cells expressed GABA responsive Cl− channels and benzodiazepine binding sites with the same pharmacological specificity as animal cells expressing these receptor subunits.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Schofield, P.R., Darlison, M.G., Fujita, N., Burt, D.R., Stephenson, F.A., Rodriguez, H., Rhee, L.M., Ramachandran, J., Reale, V., Glencorse, T.A., Seeburg, P.H. and Barnard, E.A. 1987 Sequence and functional expiession of the GABAA receptor show a ligind gated receptor superfamily. Nature 328: 221–227.

    Article  CAS  Google Scholar 

  2. Pritchett, D.B., Sontheimer, H., Gorman, C.M., Kettenmann, H., Seeburg, P.H. and Schofield, P.R. 1988 Transient expression shows ligind gating and allosteric polentiation of GABAA receptor subumts. Science 242: 1306–1308.

    Article  CAS  Google Scholar 

  3. Pritchett, D.B., Sontheimer, H., Shivers, B.D., Ymer, S., Kettenmann, H., Schofield, P.R. and Seeburg, P.H. 1988. Importance of a novel GABAA receptor subunit for benzodiazepine pliaimacology. Nature 338: 582–585.

    Article  Google Scholar 

  4. Pritchett, D.B., L¨ddens, H. and Seehuig, P.H. 1989. Type I and type II GABAA benzodiazepine receptors produced in transfected cells. Science 245: 1389–1392.

    Article  CAS  Google Scholar 

  5. Shivers, B.D., Killisch, I., Sprengel, R., Sontheimer, H., Köhler, M., Schofield, P.R. and Seeburg, P.H. 1989. Two novel GABAA receptor subunits exist in distinct neuronal subpopulations. Neuron 3: 327–337.

    Article  CAS  Google Scholar 

  6. Ymer, S., Schofield, P.R., Draguhn, A., Werner, P., Köhler, M. and Seeburg, P.H. 1989 GABAA receptor β subunits heteiogeneity: functional expression of cloned cDNAs. EMBO J. 8: 1665–1670.

    Article  CAS  Google Scholar 

  7. Pritchett, D.B. and Seeburg, P.H. 1990. α-Aminobutyric acidA receptor α5 sub unit creates novel type II benzodiazepine receptor pharmacology. J. of Neurochem. 54: 1802–1804.

    Article  CAS  Google Scholar 

  8. Putlitz, J., Kubasek, W.L., Duchene, M., Marget, M., vonSpecht, B.U. and Domdey, H. 1990. Antibody production from baculovirus infected insect cells. Bio/Technology 8: 651–654.

    Google Scholar 

  9. Angelo, C.S., Smith, G.E., Summers, M.D. and Krug, R.M. 1987. Two of the three influenza viral polymerase proteins expressed by using baculovirus vectors form a complex in insect cells. J. Gen. Virol. 61: 361–365.

    CAS  Google Scholar 

  10. Klaiber, K., Williams, N., Roberts, T.M., Papazian, D.M., Jan, L.Y. and Miller, C. 1990. Functional expression of shaker K+ channels in a baculovirus-infected insect cell line. Neuron 5: 221–226.

    Article  CAS  Google Scholar 

  11. Puia, G., Vicini, S., Seeburg, P.H. and Costa, E. 1991. Influence of recombinant γ-aminobutryic acidA receptor subunit composition on the action of allosteric modulators of γ-aminobutryic acid-gated Cl currents. Mol. Biol. 39: 691–696.

    CAS  Google Scholar 

  12. Tallman, J.F., Thomas, J.W. and Gallager, D.W. 1978. GABAergic modulation of benzodiazepine binding site sensitivity. Nature 274: 383–385.

    Article  CAS  Google Scholar 

  13. Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold spring Harbor. N.Y.

    Google Scholar 

  14. Luckow, V.A. and Summers, M.D. 1989. High level expression of nonfused foreign genes with Autographa California nuclear polyhedrosis virus expression vector. Virology 170: 31–39.

    Article  CAS  Google Scholar 

  15. Summers, M.D. and Smith, G.E. 1987. A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures. Texas Agricultural Experimental Station Bulletin No. 1555.

  16. Cheng, Y.-C. and Prusoff, W.H. 1973. Relationship between the inhibition constant (K1) and the concentrations of inhibitor which causes 50 percent inhibition (I50) of an enzymatic reaction. Biochemical Pharmacol. 22: 3099–3108.

    Article  CAS  Google Scholar 

  17. Hamill, O.P., Marty, A., Neher, E., Sakmann, B. and Sigworth, E.J. 1981. Improved patch-clamp techniques for high resolution current recording from cells and cell-free membrane patches. Pfl¨gers Arch. 391: 85–100.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Molecular Biology Research, The Upjohn Company, Kalamazoo, MI, 49007

    D. B. Carter, D. R. Thomsen, D. J. Lennon & W. Gale

  2. CNS Research, The Upjohn Company, Kalamazoo, MI, 49007

    W. B. Im & H. K. Im

  3. Chemical and Biological Screening, The Upjohn Company, Kalamazoo, MI, 49007

    D. M. Ngo & M. W. Smith

  4. University of Heidelberg, Center for Molecular Biology, D-6900, Heidelberg, Germany

    P. H. Seeburg

Authors
  1. D. B. Carter
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. R. Thomsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. W. B. Im
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. J. Lennon
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D. M. Ngo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. W. Gale
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H. K. Im
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. P. H. Seeburg
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. M. W. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Carter, D., Thomsen, D., Im, W. et al. Functional Expression of GABAA Chloride Channels and Benzodiazepine Binding Sites in Baculovirus Infected Insect Cells. Nat Biotechnol 10, 679–681 (1992). https://doi.org/10.1038/nbt0692-679

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0692-679

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing