Assembly of functional GABAA receptors in insect cells using baculovirus expression vectors

Neuroreport. 1992 Jul;3(7):597-600. doi: 10.1097/00001756-199207000-00013.

Abstract

We have constructed recombinant baculoviruses containing cDNAs encoding either the alpha 1- or the beta 1-subunit of the bovine GABAA receptor. In Spodoptera frugiperda (IPLB-Sf-21) cells infected with recombinant virus expressing either the alpha 1- or beta 1-subunit, or in cells co-infected with both viruses, functional GABAA receptors were detected by whole-cell electrophysiological recordings. The threshold for the responses mediated by the homo-oligomeric channels (alpha- or beta-) was 2-3 x 10(-6) M GABA, and for the co-infected cells was 8 x 10(-8) M GABA, suggesting that hetero-oligomeric channels formed in these cells. All GABA-induced currents were found to be inhibited by bicuculline and picrotoxin, potentiated by pentobarbital but were insensitive to benzodiazepines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Bicuculline / pharmacology
  • Cattle
  • Cell Line
  • Cloning, Molecular
  • Evoked Potentials / drug effects
  • Genetic Vectors
  • Macromolecular Substances
  • Moths
  • Pentobarbital / pharmacology
  • Receptors, GABA-A / biosynthesis
  • Receptors, GABA-A / genetics
  • Receptors, GABA-A / physiology*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / metabolism
  • Transfection*
  • Virulence Factors, Bordetella / pharmacology
  • gamma-Aminobutyric Acid / pharmacology*

Substances

  • Macromolecular Substances
  • Receptors, GABA-A
  • Recombinant Proteins
  • Virulence Factors, Bordetella
  • gamma-Aminobutyric Acid
  • Pentobarbital
  • Bicuculline