Effect of N′-nitrosonornicotine (NNN) on murine palatal fusion in vitro

Takashi Saito, Xiao Mei Cui, Tadashi Yamamoto, Nobuyuki Shiomi, Pablo Bringas, Charles F. Shuler

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Maternal smoking has been linked to an increased risk for orofacial clefts. N′-nitrosonornicotine (NNN) is one of the tobacco-specific nitrosamines that has been shown to be linked to the deleterious effects of tobacco and could be linked to the formation of cleft palate birth defects. The effect of NNN on palatal fusion was examined using an in vitro organ culture model of palatal development. The organ cultures were exposed to NNN (0.01, 0.1, 1, 10 and100 mM) and the effects on palatal development characterized at defined points. Palatal fusion was evaluated at embryonic day 13 (E13) + 72 h by characterizing the remaining medial edge epithelium (MEE) and determining the extent of fusion compared to controls. The NNN-treated group (1 mM) had more MEE remaining in the palatal midline than the untreated group at E13 + 72 h (P < 0.05). Changes in cell proliferation in the MEE resulting from NNN exposure were examined by BrdU incorporation in replicating DNA. Changes in the pattern of MEE cell death were examined by TUNEL. BrdU incorporation and TUNEL staining showed that the NNN (1 mM)-treated palates had more MEE cell proliferation and less apoptosis than the untreated-palates at E13 + 24 h (P < 0.05). The mechanism altered by NNN was further evaluated by characterizations of extracellular signal-regulated kinase (ERK) 1/2, p38 and c-jun amino-terminal kinase (JNK). NNN at 1 mM induced ERK1/2 phosphorylation, but reduced p38 phosphorylation (P < 0.05, P < 0.01, respectively) in the MEE. The results suggest that NNN inhibited palatal fusion by effects on cell proliferation and MEE cell death.

Original languageEnglish
Pages (from-to)475-485
Number of pages11
JournalToxicology
Volume207
Issue number3
DOIs
Publication statusPublished - Feb 28 2005
Externally publishedYes

Fingerprint

N'-nitrosonornicotine
Fusion reactions
Epithelium
Cell proliferation
Phosphorylation
Tobacco
Palate
Organ Culture Techniques
Cell Proliferation
In Situ Nick-End Labeling
Cell death
Bromodeoxyuridine
Cell Death
In Vitro Techniques
Nitrosamines
Mitogen-Activated Protein Kinase 3
JNK Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinase 1

Keywords

  • Cell proliferation
  • Medial edge epithelium
  • N′-nitrosonornicotine
  • Palate
  • Programmed cell death
  • Tobacco

ASJC Scopus subject areas

  • Toxicology

Cite this

Saito, T., Cui, X. M., Yamamoto, T., Shiomi, N., Bringas, P., & Shuler, C. F. (2005). Effect of N′-nitrosonornicotine (NNN) on murine palatal fusion in vitro. Toxicology, 207(3), 475-485. https://doi.org/10.1016/j.tox.2004.10.015

Effect of N′-nitrosonornicotine (NNN) on murine palatal fusion in vitro. / Saito, Takashi; Cui, Xiao Mei; Yamamoto, Tadashi; Shiomi, Nobuyuki; Bringas, Pablo; Shuler, Charles F.

In: Toxicology, Vol. 207, No. 3, 28.02.2005, p. 475-485.

Research output: Contribution to journalArticle

Saito, T, Cui, XM, Yamamoto, T, Shiomi, N, Bringas, P & Shuler, CF 2005, 'Effect of N′-nitrosonornicotine (NNN) on murine palatal fusion in vitro', Toxicology, vol. 207, no. 3, pp. 475-485. https://doi.org/10.1016/j.tox.2004.10.015
Saito, Takashi ; Cui, Xiao Mei ; Yamamoto, Tadashi ; Shiomi, Nobuyuki ; Bringas, Pablo ; Shuler, Charles F. / Effect of N′-nitrosonornicotine (NNN) on murine palatal fusion in vitro. In: Toxicology. 2005 ; Vol. 207, No. 3. pp. 475-485.
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