Acetylcholine and alcohol sensitivity of neuronal nicotinic acetylcholine receptors: Mutations in transmembrane domains

Cecilia M. Borghese, Deeba N. Ali, Virginia Bleck, R. Adron Harris

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Background: The effect of n-alcohols on glycine and γ-aminobutyric acid type A receptors depends on two specific amino acids (AAs) located in the transmembrane domains (TM) 2 and 3. Our aim was to assess whether the corresponding AAs in the neuronal nicotinic acetylcholine receptor (nAChR) also formed a binding pocket for alcohols. Methods: We made single AA substitutions in the homologous sites in rat neuronal nAChR α2 and α4 (αL261 and αL283) and expressed them in Xenopus laevis oocytes in combination with β4 wild type. The effect of different n-alcohols was studied in α4(L261A)β4 and α4(L283A)β4 nAChRs. The effect of ethanol, propanol, and octanol on acetylcholine (ACh) responses was studied in α2(L261X)β4 and α2(L283X)β4 nAChRs. Results: Most of the mutations in the α2 subunit, in either the 261 or the 283 position, induced changes in ACh sensitivity and increased alcohol action, but none was able to reduce ethanol potentiation. In α4(L283A)β4, enhancement of potentiation by short-chain alcohols was observed, as well as a change from inhibition to potentiation for long-chain alcohols. The exposure of the AAs was assessed through the action of a charged thiol-specific reagent on α2(L261C)β4 and α2(L283C)β4, and these experiments suggest that the AA in TM2 is located in a water-accessible position, whereas the AA in TM3 is inaccessible. However, a noncharged thiol-specific reagent did not affect either ACh responses or ethanol effect on α2(L261C)β4. Conclusions: The AAs located at positions 261 and 283 of the α2 and α4 nAChR subunits do not seem to form a binding pocket for alcohols. Additional studies are required to determine whether alcohols act on a site near these AAs or on sites unrelated to the TM2-TM3 site found in glycine and γ-aminobutyric acid type A receptors.

Original languageEnglish (US)
Pages (from-to)1764-1772
Number of pages9
JournalAlcoholism: Clinical and Experimental Research
Volume26
Issue number12
DOIs
StatePublished - Dec 1 2002

Keywords

  • Alcohol
  • Ligand-Gated Ion Channel
  • Nicotinic Acetylcholine Receptor
  • Transmembrane Domain
  • Xenopus Laevis Oocyte

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Toxicology
  • Psychiatry and Mental health

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