Immortalized hypothalamic GT1-7 neurons express functional gamma-aminobutyric acid type A receptors

T G Hales, H Kim, B Longoni, R W Olsen, A J Tobin

Research output: Contribution to journalArticlepeer-review

70 Citations (Scopus)

Abstract

Neuronal cell lines provide a source of pure populations of neurons and allow the properties of many neurotransmitter receptors to be studied. However, none of these cells have been reported to express functional gamma-aminobutyric acid (GABA)A receptors. Indeed, there have been no reports of cell lines expressing functional amino acid receptors. Using biochemical and electrophysiological techniques, we have identified a neuronal cell line expressing functional GABAA receptors. Membranes from immortalized hypothalamic (GT1-7) neurons bound [3H]muscimol but not [3H]flunitrazepam. GABA-activated chloride currents, recorded from GT1-7 cells, were blocked by bicuculline and Zn2+ but were insensitive to diazepam. These results suggest that GABAA receptors on GT1-7 cells lack gamma subunits. The neurosteroid 5 alpha-pregnan-3 alpha-ol-20-one and pentobarbital both modulated GABAA receptors in these cells. Polymerase chain reaction analysis of the cells revealed the presence of mRNAs encoding alpha 1, beta 1, and beta 3 polypeptides. GT1-7 cells provide a useful model system for studying the regulation of GABAA receptor polypeptide expression.

Original languageEnglish
Pages (from-to)197-202
Number of pages6
JournalMolecular Pharmacology
Volume42
Issue number2
Publication statusPublished - Aug 1992

Keywords

  • Anesthetics/pharmacology
  • Animals
  • Barbiturates/pharmacology
  • Base Sequence
  • Bicuculline/pharmacology
  • Cell Line, Transformed
  • Cell Membrane/metabolism
  • Chloride Channels
  • Diazepam/pharmacology
  • Electrophysiology
  • Flunitrazepam/pharmacology
  • GABA Antagonists
  • Gene Expression Regulation/genetics
  • Hypothalamus/pathology
  • Membrane Proteins/physiology
  • Mice
  • Molecular Sequence Data
  • Muscimol/metabolism
  • Neurons/pathology
  • Peptides/genetics
  • Polymerase Chain Reaction
  • RNA, Messenger/analysis
  • Receptors, GABA-A/classification
  • Sensitivity and Specificity
  • Steroids/pharmacology
  • Tritium
  • Tumor Cells, Cultured
  • Zinc/pharmacology
  • gamma-Aminobutyric Acid/pharmacology

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