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Volume 17, Number 4, Issue of February 15, 1997 pp. 1350-1362
Copyright ©1997 Society for Neuroscience

Ligand-Gated Ion Channel Subunit Partnerships: GABAA Receptor alpha 6 Subunit Gene Inactivation Inhibits delta  Subunit Expression

Received Oct. 10, 1996; revised Nov. 25, 1996; accepted Nov. 26, 1996.

A. Jones1, , E. R. Korpi2, , R. M. McKernan3, , R. Pelz4, , Z. Nusser5, R. Mäkelä2, J. R. Mellor1, S. Pollard6, S. Bahn1, F. A. Stephenson6, A. D. Randall1, W. Sieghart4, P. Somogyi5, A. J. H. Smith1, and W. Wisden1

1 Medical Research Council Laboratory of Molecular Biology, Medical Research Council Centre, Cambridge CB2 2QH, United Kingdom, 2 Department of Pharmacology and Clinical Pharmacology, University of Turku, Fin-20014 Turku, Finland, Department of Alcohol Research, National Public Health Institute, Fin-00180 Helsinki, Finland, 3 Merck Sharp & Dohme Research Laboratories, Neuroscience Research Centre, Harlow, CM20 2QR, United Kingdom, 4 University Clinic for Psychiatry, Department of Biochemical Psychiatry, 1090 Vienna, Austria, 5 Medical Research Council Anatomical Neuropharmacology Unit, Oxford University Department of Pharmacology, Oxford, OX1 3TH, United Kingdom, and 6 School of Pharmacy, University of London, Brunswick Square, London, WC1N 1AX, United Kingdom

Cerebellar granule cells express six GABAA receptor subunits abundantly (alpha 1, alpha 6, beta 2, beta 3, gamma 2, and delta ) and assemble various pentameric receptor subtypes with unknown subunit compositions; however, the rules guiding receptor subunit assembly are unclear. Here, removal of intact alpha 6 protein from cerebellar granule cells allowed perturbations in other subunit levels to be studied. Exon 8 of the mouse alpha 6 subunit gene was disrupted by homologous recombination. In alpha 6 -/- granule cells, the delta  subunit was selectively degraded as seen by immunoprecipitation, immunocytochemistry, and immunoblot analysis with delta  subunit-specific antibodies. The delta  subunit mRNA was present at wild-type levels in the mutant granule cells, indicating a post-translational loss of the delta  subunit. These results provide genetic evidence for a specific association between the alpha 6 and delta  subunits. Because in alpha 6 -/- neurons the remaining alpha 1, beta 2/3, and gamma 2 subunits cannot rescue the delta  subunit, certain potential subunit combinations may not be found in wild-type cells.

Key words: GABAA receptor; alpha 6 subunit; granule cell; cerebellum; homologous recombination; gene targeting; transgenic mice; knockout mice; ligand-gated ion channel; subunit sorting; subunit assembly; internal ribosome entry site; dicistronic mRNA; muscimol; SR95531; Ro 15-4513; flunitrazepam




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Z. Nusser, W. Sieghart, and P. Somogyi
Segregation of Different GABAA Receptors to Synaptic and Extrasynaptic Membranes of Cerebellar Granule Cells
J. Neurosci., March 1, 1998; 18(5): 1693 - 1703.
[Abstract] [Full Text] [PDF]

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 Mol. Pharmacol.Home page
R. Mäkelä, M. Uusi-Oukari, G. E. Homanics, J. J. Quinlan, L. L. Firestone, W. Wisden, and E. R. Korpi
Cerebellar gamma -Aminobutyric Acid Type A Receptors: Pharmacological Subtypes Revealed by Mutant Mouse Lines
Mol. Pharmacol., September 1, 1997; 52(3): 380 - 388.
[Abstract] [Full Text]

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 Proc. Natl. Acad. Sci. USAHome page
S. Bahn, A. Jones, and W. Wisden
Directing gene expression to cerebellar granule cells using gamma -aminobutyric acid type A receptor alpha 6 subunit transgenes
PNAS, August 19, 1997; 94(17): 9417 - 9421.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
V. Tretter, B. Hauer, Z. Nusser, R. M. Mihalek, H. Hoger, G. E. Homanics, P. Somogyi, and W. Sieghart
Targeted Disruption of the GABAA Receptor delta Subunit Gene Leads to an Up-regulation of gamma 2 Subunit-containing Receptors in Cerebellar Granule Cells
J. Biol. Chem., March 23, 2001; 276(13): 10532 - 10538.
[Abstract] [Full Text] [PDF]


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