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

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

Research output: Contribution to journalArticle

47 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Nicotinic Receptors
Tetracycline
Nicotine
Up-Regulation
Bungarotoxins
Tubocurarine
Carbachol
Neuroblastoma
Transgenes
Membrane Proteins
Epithelial Cells
Pharmacology
Cell Line
Messenger RNA

Keywords

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

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Inducible, heterologous expression of human α7-nicotinic acetylcholine receptors in a native nicotinic receptor-null human clonal line. / Peng, Jian Hong; Lucero, Linda; Fryer, John D.; Herl, Jennifer; Leonard, Sherry S.; Lukas, Ronald J.

In: Brain Research, Vol. 825, No. 1-2, 17.04.1999, p. 172-179.

Research output: Contribution to journalArticle

Peng, Jian Hong ; Lucero, Linda ; Fryer, John D. ; Herl, Jennifer ; Leonard, Sherry S. ; Lukas, Ronald J. / Inducible, heterologous expression of human α7-nicotinic acetylcholine receptors in a native nicotinic receptor-null human clonal line. In: Brain Research. 1999 ; Vol. 825, No. 1-2. pp. 172-179.
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