Nicotine facilitates nicotinic acetylcholine receptor targeting to mitochondria but makes them less susceptible to selective ligands

Kateryna Uspenska, Olena Lykhmus, Galyna Gergalova, Volodymyr Chernyshov, Hugo R. Arias, Sergiy Komisarenko, Maryna Skok

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Several nicotinic acetylcholine receptor (nAChR) subtypes are expressed in mitochondria to regulate the internal pathway of apoptosis in ion channel-independent manner. However, the mechanisms of nAChR activation in mitochondria and targeting to mitochondria are still unknown. Nicotine has been shown to favor nAChR pentamer assembly, folding, and maturation on the way of biosynthesis. The idea of the present work was to determine whether nicotine affects the content, glycosylation, and function of mitochondrial nAChRs. Experiments were performed in isolated liver mitochondria from mice, that either consumed or not nicotine with the drinking water (200 μL/L) for 7 days. Mitochondria detergent lysates were studied by sandwich or lectin ELISA for the presence and carbohydrate composition of different nAChR subunits. Intact mitochondria were examined by flow cytometry for the binding of fluorescently labeled α-cobratoxin and were tested in functional assay of cytochrome c release under the effect of either Ca2+ or wortmannin in the presence or absence of nAChR-selective ligands, including PNU-282987 (1 nM), dihydro-β-erythroidine (DhβE, 1 μM), PNU-120596 (0.3, 3, or 10 μM) and desformylflustrabromine hydrochloride (dFBr, 0.001, 0.3, or 1 μM). It was found that nicotine consumption increased the ratio of mitochondrial vs non-mitochondrial nAChRs in the liver, enhanced fucosylation of mitochondrial nAChRs, but prevented the binding of α-cobratoxin and the cytochrome c release-attenuating effects of nAChR-specific agonists, antagonists, or positive allosteric modulators. It is concluded that nicotine consumption in vivo favors nAChR glycosylation and trafficking to mitochondria but makes them less susceptible to the effects of specific ligands.

Original languageEnglish
Pages (from-to)43-50
Number of pages8
JournalNeuroscience Letters
Volume656
DOIs
StatePublished - 24 Aug 2017
Externally publishedYes

Fingerprint

Nicotinic Receptors
Nicotine
Mitochondria
Ligands
Cytochromes c
Glycosylation
Liver Mitochondrion
Ion Channels
Lectins
Drinking Water
Detergents
Flow Cytometry
Enzyme-Linked Immunosorbent Assay
Carbohydrates
Apoptosis
Liver

Keywords

  • Cytochrome c
  • Glycosylation
  • Mitochondria
  • Nicotine
  • Nicotinic acetylcholine receptor

Cite this

Uspenska, Kateryna ; Lykhmus, Olena ; Gergalova, Galyna ; Chernyshov, Volodymyr ; Arias, Hugo R. ; Komisarenko, Sergiy ; Skok, Maryna. / Nicotine facilitates nicotinic acetylcholine receptor targeting to mitochondria but makes them less susceptible to selective ligands. In: Neuroscience Letters. 2017 ; Vol. 656. pp. 43-50.
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Nicotine facilitates nicotinic acetylcholine receptor targeting to mitochondria but makes them less susceptible to selective ligands. / Uspenska, Kateryna; Lykhmus, Olena; Gergalova, Galyna; Chernyshov, Volodymyr; Arias, Hugo R.; Komisarenko, Sergiy; Skok, Maryna.

In: Neuroscience Letters, Vol. 656, 24.08.2017, p. 43-50.

Research output: Contribution to journalArticle

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T1 - Nicotine facilitates nicotinic acetylcholine receptor targeting to mitochondria but makes them less susceptible to selective ligands

AU - Uspenska, Kateryna

AU - Lykhmus, Olena

AU - Gergalova, Galyna

AU - Chernyshov, Volodymyr

AU - Arias, Hugo R.

AU - Komisarenko, Sergiy

AU - Skok, Maryna

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AB - Several nicotinic acetylcholine receptor (nAChR) subtypes are expressed in mitochondria to regulate the internal pathway of apoptosis in ion channel-independent manner. However, the mechanisms of nAChR activation in mitochondria and targeting to mitochondria are still unknown. Nicotine has been shown to favor nAChR pentamer assembly, folding, and maturation on the way of biosynthesis. The idea of the present work was to determine whether nicotine affects the content, glycosylation, and function of mitochondrial nAChRs. Experiments were performed in isolated liver mitochondria from mice, that either consumed or not nicotine with the drinking water (200 μL/L) for 7 days. Mitochondria detergent lysates were studied by sandwich or lectin ELISA for the presence and carbohydrate composition of different nAChR subunits. Intact mitochondria were examined by flow cytometry for the binding of fluorescently labeled α-cobratoxin and were tested in functional assay of cytochrome c release under the effect of either Ca2+ or wortmannin in the presence or absence of nAChR-selective ligands, including PNU-282987 (1 nM), dihydro-β-erythroidine (DhβE, 1 μM), PNU-120596 (0.3, 3, or 10 μM) and desformylflustrabromine hydrochloride (dFBr, 0.001, 0.3, or 1 μM). It was found that nicotine consumption increased the ratio of mitochondrial vs non-mitochondrial nAChRs in the liver, enhanced fucosylation of mitochondrial nAChRs, but prevented the binding of α-cobratoxin and the cytochrome c release-attenuating effects of nAChR-specific agonists, antagonists, or positive allosteric modulators. It is concluded that nicotine consumption in vivo favors nAChR glycosylation and trafficking to mitochondria but makes them less susceptible to the effects of specific ligands.

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