Promoted Neuronal Differentiation after Activation of Alpha4/Beta2 Nicotinic Acetylcholine Receptors in Undifferentiated Neural Progenitors

Takeshi Takarada, Noritaka Nakamichi, Seiya Kitajima, Ryo Fukumori, Ryota Nakazato, Nguyen Quynh Le, Yeong Hun Kim, Koichi Fujikawa, Miki Kou, Yukio Yoneda

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Background: Neural progenitor is a generic term used for undifferentiated cell populations of neural stem, neuronal progenitor and glial progenitor cells with abilities for proliferation and differentiation. We have shown functional expression of ionotropic N-methyl-D-aspartate (NMDA) and gamma-aminobutyrate type-A receptors endowed to positively and negatively regulate subsequent neuronal differentiation in undifferentiated neural progenitors, respectively. In this study, we attempted to evaluate the possible functional expression of nicotinic acetylcholine receptor (nAChR) by undifferentiated neural progenitors prepared from neocortex of embryonic rodent brains. Methodology/Principal Findings: Reverse transcription polymerase chain reaction analysis revealed mRNA expression of particular nAChR subunits in undifferentiated rat and mouse progenitors prepared before and after the culture with epidermal growth factor under floating conditions. Sustained exposure to nicotine significantly inhibited the formation of neurospheres composed of clustered proliferating cells and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction activity at a concentration range of 1 μM to 1 mM without affecting cell survival. In these rodent progenitors previously exposed to nicotine, marked promotion was invariably seen for subsequent differentiation into cells immunoreactive for a neuronal marker protein following the culture of dispersed cells under adherent conditions. Both effects of nicotine were significantly prevented by the heteromeric α4β2 nAChR subtype antagonists dihydro-β-erythroidine and 4-(5-ethoxy-3-pyridinyl)-N-methyl-(3E)-3-buten-1-amine, but not by the homomeric α7 nAChR subtype antagonist methyllycaconitine, in murine progenitors. Sustained exposure to nicotine preferentially increased the expression of Math1 among different basic helix-loop-helix proneural genes examined. In undifferentiated progenitors from embryonic mice defective of NMDA receptor subunit-1, nicotine was still effective in significantly inhibiting the proliferation. Conclusions/Significance: Functional α4β2 nAChR subtype would be constitutively expressed to play a role in the mechanism underlying the determination of proliferation and subsequent differentiation fate into a neuronal lineage in association with preferential promotion of Math1 expression in undifferentiated neural progenitors of developing rodent neocortex independently of NMDA receptor activation.

Original languageEnglish
Article numbere46177
JournalPLoS One
Volume7
Issue number10
DOIs
Publication statusPublished - Oct 4 2012
Externally publishedYes

Fingerprint

cholinergic receptors
nicotine
Nicotinic Receptors
Nicotine
Chemical activation
aspartic acid
neocortex
Rodentia
rodents
Neocortex
N-Methyl-D-Aspartate Receptors
receptors
antagonists
mice
Cells
Aminobutyrates
Neural Stem Cells
Polymerase chain reaction
epidermal growth factor
neuroglia

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Promoted Neuronal Differentiation after Activation of Alpha4/Beta2 Nicotinic Acetylcholine Receptors in Undifferentiated Neural Progenitors. / Takarada, Takeshi; Nakamichi, Noritaka; Kitajima, Seiya; Fukumori, Ryo; Nakazato, Ryota; Le, Nguyen Quynh; Kim, Yeong Hun; Fujikawa, Koichi; Kou, Miki; Yoneda, Yukio.

In: PLoS One, Vol. 7, No. 10, e46177, 04.10.2012.

Research output: Contribution to journalArticle

Takarada, T, Nakamichi, N, Kitajima, S, Fukumori, R, Nakazato, R, Le, NQ, Kim, YH, Fujikawa, K, Kou, M & Yoneda, Y 2012, 'Promoted Neuronal Differentiation after Activation of Alpha4/Beta2 Nicotinic Acetylcholine Receptors in Undifferentiated Neural Progenitors', PLoS One, vol. 7, no. 10, e46177. https://doi.org/10.1371/journal.pone.0046177
Takarada, Takeshi ; Nakamichi, Noritaka ; Kitajima, Seiya ; Fukumori, Ryo ; Nakazato, Ryota ; Le, Nguyen Quynh ; Kim, Yeong Hun ; Fujikawa, Koichi ; Kou, Miki ; Yoneda, Yukio. / Promoted Neuronal Differentiation after Activation of Alpha4/Beta2 Nicotinic Acetylcholine Receptors in Undifferentiated Neural Progenitors. In: PLoS One. 2012 ; Vol. 7, No. 10.
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