Elsevier

Neuroscience

Volume 109, Issue 2, 28 January 2002, Pages 275-285
Neuroscience

Stimulation of non-α7 nicotinic receptors partially protects dopaminergic neurons from 1-methyl-4-phenylpyridinium-induced toxicity in culture

https://doi.org/10.1016/S0306-4522(01)00488-2Get rights and content

Abstract

Previous work has shown that nicotine treatment protects against nigrostriatal degeneration in rodents, findings that may be of relevance to the decreased incidence of Parkinson’s disease in cigarette smokers. In the present studies, we investigated the effect of nicotine against 1-methyl-4-phenylpyridinium-induced toxicity in dopaminergic ventral mesencephalic cultures to identify the nicotinic receptor population that may be involved. [3H]Epibatidine, a ligand that binds to receptors containing α2–α6 subunits, bound to at least two populations of sites that were up-regulated by nicotine in a time and dose dependent manner. We next examined the effect of nicotine on cultures treated with 1-methyl-4-phenylpyridinium, a neurotoxin that selectively damages nigrostriatal dopaminergic neurons. Pre-treatment with nicotine, at 10−7–10−4 M, partially prevented the toxin-induced decline in dopaminergic cells. Pre-exposure to nicotine for 24 h resulted in optimal protection, suggesting that receptor up-regulation may contribute to the observed neuroprotective effect. Nicotine-mediated protection was blocked by pre-incubation with the nicotinic receptor antagonist d-tubocurarine (10−4 M), but not the α7 receptor-selective antagonist α-bungarotoxin (10−7 M).

Our results show that nicotinic receptor activation partially protects nigral dopaminergic neurons from 1-methyl-4-phenylpyridinium-induced toxicity in culture and that this appears to occur through an interaction at non-α7 containing receptors.

Section snippets

Materials

Dulbecco’s modified Eagle’s medium (DMEM), Hank’s balanced salt solutions (HBSS), trypsin–EDTA, fetal calf serum, and penicillin/streptomycin were purchased from Life Technologies (Rockville, MD, USA); normal goat serum, Vectastain ABC kit, and Vector VIP kit from Vector Laboratories (Burlingame, CA, USA); nicotine, d-tubocurarine, α-bungarotoxin, MPP+, poly-D-lysine and polyethyleneimine from Sigma Chemical (St. Louis, MO, USA); tyrosine hydroxylase (TH) antibody from Pel-Freez Biologicals

[3H]Epibatidine binding in culture

VM cells were grown in culture for up to 2 weeks. Specific [3H]epibatidine binding was observed in these cultures as early as 3 days after plating at which time cells had begun to extend processes and form a network. Binding increased with time in culture up to 7 days after plating and then plateaued until 10 days in culture (Fig. 1A). By 14 days in culture, binding had decreased to the level seen at day 3 and the cultures appeared less viable, although the results at this time point were very

Discussion

The present results show that nicotine partially protects against MPP+-induced toxicity in mesencephalic cultures. The degree of neuroprotection is similar to that observed in in vivo studies which demonstrate ∼20% protection against nigrostriatal degeneration after nicotine or cytisine administration (Janson et al., 1988a, Janson et al., 1988b, Fuxe et al., 1990, Janson et al., 1991, Janson and Moller, 1993, Ferger et al., 1998). The similarity in the magnitude of the protective effect against

Conclusion

The results show that stimulation of non-α7 nicotinic receptors partially protects dopaminergic mesencephalic neurons from MPP+-induced toxicity in culture. These results may suggest that nicotinic receptors represent a potential therapeutic target for delaying or preventing Parkinson’s disease. The symptoms of Parkinson’s disease do not manifest until nigrostriatal degeneration has progressed to about 70–80%; therefore, even a modest increase in the integrity of nigral dopaminergic neurons may

Acknowledgements

We thank Namitha Devendrarao for excellent technical assistance. This work was supported by the Tobacco Related Disease Research Program, #8RT-0105 and #7FT-0010.

References (76)

  • B Ferger et al.

    Effects of cytisine on hydroxyl radicals in vitro and MPTP-induced dopamine depletion in vivo

    Eur. J. Pharmacol.

    (1998)
  • A.M Janson et al.

    Chronic nicotine treatment counteracts nigral cell loss induced by a partial mesodiencephalic hemitransection: an analysis of the total number and mean volume of neurons and glia in substantia nigra of the male rat

    Neuroscience

    (1993)
  • A.M Janson et al.

    Chronic nicotine treatment counteracts the disappearance of tyrosine-hydroxylase-immunoreactive nerve cell bodies, dendrites and terminals in the mesostriatal dopamine system of the male rat after partial hemitransection

    Brain Res.

    (1988)
  • A.M Janson et al.

    Chronic nicotine treatment counteracts the decrease in extracellular neostriatal dopamine induced by a unilateral transection at the mesodiencephalic junction in rats: a microdialysis study

    Neurosci. Lett.

    (1991)
  • S Kaneko et al.

    Nicotine protects cultured cortical neurons against glutamate-induced cytotoxicity via alpha7-neuronal receptors and neuronal CNS receptors

    Brain Res.

    (1997)
  • J Komourian et al.

    Characterization of nicotinic receptors in immortalized hippocampal neurons

    Brain Res.

    (1996)
  • Y Li et al.

    alpha7 nicotinic receptor mediated protection against ethanol-induced cytotoxicity in PC12 cells

    Brain Res.

    (1999)
  • Y Li et al.

    Characterization of the neuroprotective and toxic effects of α7 nicotinic receptor activation in PC12 cells

    Brain Res.

    (1999)
  • M.D Minana et al.

    Nicotine prevents glutamate-induced proteolysis of the microtubule-associated protein MAP-2 and glutamate neurotoxicity in primary cultures of cerebellar neurons

    Neuropharmacology

    (1998)
  • M Quik et al.

    Alteration in nicotinic receptor subunit mRNAs in monkey brain after nigrostriatal degeneration

    Neuroscience

    (2000)
  • G.S Shahi et al.

    1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity: partial protection against striato-nigral dopamine depletion in C57BL/6J mice by cigarette smoke exposure and by beta-naphthoflavone pretreatment

    Neurosci. Lett.

    (1991)
  • E.M Sorenson et al.

    Postsynaptic nicotinic receptors on dopaminergic neurons in the substantia nigra pars compacta of the rat

    Neuroscience

    (1998)
  • S Wonnacott

    Presynaptic nicotinic Ach receptors

    Trends Neurosci.

    (1997)
  • M Arroyo-Jimenez et al.

    Ultrastructural localization of the α4-subunit of the neuronal acetylcholine nicotinic receptor in the rat substantia nigra

    J. Neurosci.

    (1999)
  • L Azam et al.

    Expression of neuronal nicotinic acetylcholine receptor subunit mRNAs within rat midbrain dopaminergic neurons

    Soc. Neurosci. Abstr.

    (1999)
  • J.A Baron

    Cigarette smoking and Parkinson’s disease

    Neurology

    (1986)
  • J.A Baron

    Beneficial effects of nicotine and cigarette smoking: The real, the possible and the spurious

    Br. Med. Bull.

    (1996)
  • R.A Behmand et al.

    Nicotine enhances 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine neurotoxicity

    J. Neurochem.

    (1992)
  • N Belluardo et al.

    The nicotinic acetylcholine receptor agonist ABT-594 increases FGF-2 expression in various rat brain regions

    NeuroReport

    (1999)
  • M.E.M Benwell et al.

    Evidence that tobacco smoking increases the density of 3H-nicotine binding sites in human brain

    J. Neurochem.

    (1988)
  • C.R Breese et al.

    Effect of smoking history on 3H-nicotine binding in human postmortem brain

    J. Pharmacol. Exp. Ther.

    (1997)
  • A Brouard et al.

    Mesencephalic dopaminergic neurons in primary cultures express functional neurotensin receptors

    J. Neurosci.

    (1992)
  • D Casper et al.

    EGF enhances the survival of dopamine neurons in rat embryonic mesencephalon primary cell culture

    J. Neurosci. Res.

    (1991)
  • E Charpantier et al.

    Nicotinic acetylcholine subunit mRNA expression in dopaminergic neurons of the rat substantia nigra and ventral tegmental area

    NeuroReport

    (1998)
  • P.B Clarke et al.

    Nicotinic binding in rat brain: autoradiographic comparison of [3H]acetylcholine, [3H]nicotine, and [125I]-α-bungarotoxin

    J. Neurosci.

    (1985)
  • A.C Collins et al.

    Sensitivity to nicotine and brain nicotinic receptors are altered by chronic nicotine and mecamylamine infusion

    J. Pharmacol. Exp. Ther.

    (1994)
  • K.J Elliott et al.

    6-hydroxydopamine lesion of rat nigrostriatal dopaminergic neurons differentially affects nicotinic acetylcholine receptor subunit mRNA expression

    J. Mol. Neurosci.

    (1998)
  • K Fuxe et al.

    Chronic nicotine treatment increases dopamine levels and reduces dopamine utilization in substantia nigra and in surviving forebrain dopamine nerve terminal systems after a partial di-mesencephalic hemitransection

    Naunyn Schmiedebergs Arch. Pharmacol.

    (1990)
  • Cited by (70)

    • Characterization of AN317, a novel selective agonist of α6β2-containing nicotinic acetylcholine receptors

      2020, Biochemical Pharmacology
      Citation Excerpt :

      These results demonstrate that direct activation of α6β2* nAChRs can regulate neuronal activity in a subpopulation of dopaminergic cells in the SNc. Nicotine has previously been demonstrated to mediate neuroprotective effects on primary dopaminergic neurons in mesencephalic cultures injured by various neurotoxins [9,65,66]. To assess the putative protection against dopaminergic neurodegeneration offered by selective α6β2* nAChR activation and compare it to that mediated by non-selective stimulation of nAChR transmission, we studied the effects of AN317 and nicotine on MPP+-induced neurotoxicity on dopaminergic neurons in rat primary mesencephalic cultures.

    • Facilitatory effect of dopamine on neuromuscular transmission mediated via dopamine D1-like receptors and prospective interaction with nicotine

      2015, European Journal of Pharmacology
      Citation Excerpt :

      Compelling evidence has accumulated regarding the neuroprotective effects of nicotine (Quik et al., 2006) against central dopaminergic neurons' damage. In addition, several beneficial functional interactions have been identified in the central nervous system between dopamine and nicotine (Jeyarasasingam et al., 2002). These findings prompted us to explore if similar interactions exist at the level of the motor end plate, how these interactions affect neuromuscular transmission and skeletal muscle performance and could these interactions be the target for new prospective treatment of motor deficit syndromes.

    • Nicotine and Nicotinic Receptor Drugs: Potential for Parkinson's Disease and Drug-Induced Movement Disorders

      2015, International Review of Neurobiology
      Citation Excerpt :

      More importantly, extensive studies showed that nicotine and nAChR agonist administration protected against nigrostriatal damage in parkinsonian monkey, rat, and mouse models (Dajas, Costa, Abin-Carriquiry, McGregor, & Urbanavicius, 2001; Janson et al., 1988; Maggio et al., 1998, 1997; Mudo, Belluardo, Mauro, & Fuxe, 2007; Quik, O'Neill, et al., 2007; Quik et al., 2006; Shimohama, 2009; Stuckenholz et al., 2013; Suzuki et al., 2013). Studies with genetically modified mice and selective nAChR drugs indicate that protection is mediated by various nAChR populations, including the β2* (with the asterisk indicating the presence of other nAChR subunits in the receptor complex) and α7 subtypes (Bordia et al., 2015; Jeyarasasingam, Tompkins, & Quik, 2002; Park et al., 2007; Ryan, Ross, Drago, & Loiacono, 2001; Stuckenholz et al., 2013). These preclinical studies formed the basis for a Michael J. Fox Foundation funded clinical trial, currently in progress, to investigate the potential of the nicotine patch to protect against disease progression in early onset Parkinson's disease (ClinicalTrials.gov Identifier NCT01560754).

    • Secalonic acid A protects dopaminergic neurons from 1-methyl-4- phenylpyridinium (MPP<sup>+</sup>)-induced cell death via the mitochondrial apoptotic pathway

      2013, European Journal of Pharmacology
      Citation Excerpt :

      The Meta Morph software (version 5.0) was used to measure the TH-positive neurons and neurites. According to previous descriptions (Akaneya et al., 1995; Madsen et al., 2003; Shimizu et al., 2003; Jeyarasasingam et al., 2002), neurons with an identifiable nucleus showing at least one process with a minimum length of two times the diameter of the nucleus were recognized as TH- positive neurons. To quantify the number of TH-positive neurons, 29 representative areas per well were counted.

    • Association of nicotine dependence susceptibility gene, CHRNA5, with Parkinson's disease age at onset: Gene and smoking status interaction

      2013, Parkinsonism and Related Disorders
      Citation Excerpt :

      It is possible that the delay in AAO among smokers carrying the ND risk variants is mediated by increased consumption of nicotine associated with rs588765 genotype. Nicotine has a neuroprotective effect on dopaminergic cells, as shown in cultured cells and animal models [30–32]. The 15q24 cluster SNPs are associated not only with ND but also with quantitative cigarettes consumption measurements (e.g. cigarettes per day), supporting the notion that delayed AAO is mediated by the beneficial effect of increased nicotine exposure.

    View all citing articles on Scopus
    1

    Present address: Office of Minority Health and Research, NINDS, 6001 Executive Boulevard, Suite 2149, Bethesda, MD 20892-9535, USA.

    View full text