Inducible, heterologous expression of human α7-nicotinic acetylcholine receptors in a native nicotinic receptor-null human clonal line

Jian Hong Peng, Linda Lucero, John Fryer, Jennifer Herl, Sherry S. Leonard, Ronald J. Lukas

Research output: Contribution to journalArticle

48 Scopus citations

Abstract

Tetracycline-regulated expression of recombinant nicotinic acetylcholine receptors (nAChR) composed of human α7 subunits is achieved in native nAChR- null SH-EP1 human epithelial cells. α7 subunits are heterologously expressed as messenger RNA and as components of 125I-labeled α-bungarotoxin (I- Bgt)-binding nAChR (~ 10 pmol per milligram of membrane protein) at levels sensitive to the amount of tetracycline in cell growth medium. I-Bgt-binding α7-nAChR appear on the cell surface pool and in intracellular pools. The pharmacological profile for drug competition toward I-Bgt binding to these recombinant α7-nAChR matches that of human native α7-nAChR naturally expressed in SH-SY5Y human neuroblastoma cells (rank order potency methyllycaconitine > 1,1-dimethyl-4-phenylpiperazinium > (-)nicotine > cytisine > carbamylcholine ≥ D-tubocurarine). Chronic exposure to nicotine induces up-regulation of human recombinant α7-nAChR (80% up-regulation at 10 μM nicotine) just as it does native α7-nAChR in other human cell lines. These studies confirm expression of nAChR as homooligomers of human α7 subunits from transgenes, establish a native nAChR-null background for such expression, and demonstrate that this expression can be regulated to facilitate studies of human α7-nAChR.

Original languageEnglish (US)
Pages (from-to)172-179
Number of pages8
JournalBrain Research
Volume825
Issue number1-2
DOIs
StatePublished - Apr 17 1999

Keywords

  • Acetylcholine
  • Nicotine
  • Nicotinic acetylcholine receptor
  • SH-EP1 cell
  • Tetracycline

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

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