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1992 Fiscal Year Final Research Report Summary

Study on role of protein phosphorylation in sialagogue-stimulated parotid gland.

Research Project

Project/Area Number 02454427
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Functional basic dentistry
Research InstitutionThe University of Tokushima

Principal Investigator

INOUE Hideo  Tokushima Univ., School of Dentistry, Professor, 歯学部, 教授 (30028732)

Co-Investigator(Kenkyū-buntansha) TSURUMI Chizuko  Tokushima Univ., School of Dentistry, Research Associate, 歯学部, 助手 (60236958)
ASHIDA Yoshiyuki  Tokushima Univ., School of Dentistry, Research Associate, 歯学部, 助手 (40212500)
Project Period (FY) 1990 – 1992
Keywordssialagogue / protein phosphorylation / parotid / ornithine decarboxylase / amylase secretion / proto-oncogene / mRNA / herbimycin A
Research Abstract

Changes in protein phosphorylation and mRNA levels of ornithine decarboxylase (ODC) and proto-oncogenes in sialagogue-stimulated parotid gland were investigated in rat parotid explants cultured on siliconized lens paper floating on serum-free 199 medium.
The results were summarized as follows;
1) Isoproterenol (IPR) enhanced the phosphorylation of serine/threonine residues of four proteins with apparent molecular weights of 17K, 20K, 31K and 32K and carbachol (CC) stimulated the phosphorylation of31K and 32K proteins. These protein phosphorylations preceded the increase of ODC activity. IPR-dependent ODC induction and phosphorylation of the proteins were selectively suppressed by monensin but not by polymyxin B, whereas CC-dependent ODC induction and protein phosphorylation were inhibited by polymyxin B but not by monensin. Neither monensin nor polymyxin B suppressed IPR- or CC-stimulated amylase secretion.
2) Northern/dot and Western blot analyses revealed that the sialagogues (IPR, CC, … More methoxamine) elevated the steady-state levels of ODC mRNA and ODC protein to maxima at 2 h and 6 h, respectively, after stimulation. The increases were roughly proportional to those in ODC activity, suggesting that sialagogue-dependent enzyme induction is regulated at the transcriptional level. The mRNAs of four of nine proto-oncogenes examined showed sialagogue-dependent increases to maxima at 30 min (c-fos) or 60 min (c-jun, c-myc and c-src) after stimulation. These increases were all transient, with the levels returning to the unstimulated levels within 60 min.
3) Tyrosine kinase inhibitors such as herbimycin A, genistein and methyl 2,5-dihydroxycinnamate inhibited a sialagogue-stimulated ODC induction but enhanced a sialagogue-induced amylase secretion. A tyrosine phosphatase inhibitor, Na_3 VO_4, increased ODC activity and this increase was suppressed by herbimycin A.
These results suggest that serine/threonine and tyrosine phosphorylations of proteins participate in a sialagogue-stimulated ODC induction, and that protein tyrosine phosphorylation shows an inhibitory effect on amylase secretion. Less

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] A.Ueno,K.Kikuchi,M.Nishino,M.Kawano,N.Matsumoto and H.Inoue: "Sialagogue-stimulated Protein Phosphorylation Related to Ornithine Decarboxylase Induction in Cultured Rat Parotid Explants" Arch.Oral Biol.36. 415-423 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Ueno,M.Kawano,Y.Ashida,F.Kinoshita,and H.Inoue: "Induction of ornithine decarboxylase by sialagogues in a human Parotid gland adenocarcinoma cell line" Biochem.Int.24. 137-145 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Kawano,A.Ueno,Y.Ashida,N.Matsumoto and H.Inoue: "Effects of Sialagogues on Ornithine Decarboxylase Induction and Proto-oncogene Expression in Murine Parotid Gland" J.Dent.Res.71. 1885-1890 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Ueno, K.Kikuchi, M.Nishino, M.Kawano, N.Matsumoto and H.Inoue: "Sialagogue-stimulated protein phosphorylation related to ornithine decarboxylase induction in cultured rat parotid explants." Arch. Oral Biol. 36. 415-423 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Ueno, M.Kawano, Y.Ashida, F.Kinoshita and H.Inoue: "Induction of ornithine decarboxylase by sialagogues in a human parotid gland adenocarcinoma cell line." Biochem. Int. 24. 137-145 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Kawano, A.Ueno, Y.Ashida, N.Matsumoto and H.Inoue: "Effects of sialagogues on ornithine decarboxylase induction and proto-oncogene expression in murine parotid gland." J. Dent. Res. 71. 1885-1890 (1992)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1994-03-24  

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