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The incorporation of monomethylethanolamine and dimethylethanolamine in fetal brain aggregating cell culture

Neurochemical Research volume 13, pages 1–8 (1988)Cite this article

Abstract

Fetal rat brain aggregating cell cultures were exposed to varying concentrations of [3H]monomethylethanolamine (MME) and [3H] dimethylethanolamine (DME). The rate of labeling of water-soluble compounds was more rapid and the amount of radioactivity present was greater than in the lipids. After a 72 hour incubation in the presence of millimolar concentrations of these nitrogenous bases, the major water-soluble products were the phosphorylated form of the bases. Little label was associated with the free bases or their cytidyl derivate. In the phospholipids, 97% of the radioactivity was recovered in phosphatidylmonomethylethanolamine (PMME) and 3% in phosphatidyldimethylethanolamine (PDME) or 95% in PDME and 5% in phosphatidylcholine (PC) after growth in presence of [3H]MME and [3H]DME respectively. The rate of formation of the radioactive products increased as function of the concentration of the nitrogenous base added up to 4 mM, the highest concentration employed. There was no significant difference in the pattern of labeling with cells grown in media devoid of methionine or choline. The turnover of the water-soluble metabolites was more rapid than in the phospholipids where an apparent half-life of 24 hours was calculated.

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Abbreviations

PMT:

phospholipid-N-methyltransferase

AdoMet:

S-adenosyl-L-methionine

EA:

ethanolamine

MME:

N-monomethylethanolamine

DME:

N,N-dimethylethanolamine

CH:

choline

PE:

phosphatidylethanolamine

PMME:

phosphatidylmonomethylethanolamine

PDME:

phosphatidyldimethylethanolamine

PC:

phosphatidylcholine

PS:

phosphatidylserine

CAPS:

cyclohexylaminopropane sulfonic acid

References

  1. Akesson, B. 1977. Manipulation of phospholipid polar head group composition in primary cultures of rats hepatocytes. B.B.R.C., 76:93–99.

    Google Scholar 

  2. Honma, Y., Kasukabe, T. and Mozumi, M. 1982. Modification of membrane phospholipid composition by choline analogs induces differentiation of cultured mouse leukemia cells. B.B.A. 721:83–86.

    Google Scholar 

  3. Smith, J. D. 1983. Effect of modification of membrane phospholipid composition on the activity of phosphatidylethanolamine N-methyltransferase of Tetrahymena. Arch. Bioch. Bioph., 223:193–201.

    Google Scholar 

  4. Ansell, G. B., and Spanner, S. 1962. The effect of 2-dimethylaminoethanol on brain phospholipid metabolism. J. Neurochem. 9:253–263.

    PubMed  Google Scholar 

  5. Yavin, E. 1977. Base stimulation of phospholipid metabolism in neuroblstoma cells. Biochim. Biophys. Acta, 689:278–289.

    Google Scholar 

  6. Ansell, G. B., and Chojnacki, T. 1966. The incorporation of the phosphate esters of N-substituted aminoethanols into the phospholipids of brain and liver. Biochem. J. 98:303–310.

    PubMed  Google Scholar 

  7. Yavin, E. 1985. Polar head group decarboxylatin and metylation of phospholipids: an alternate route for phosphatidylcholine formation in cultured neuronal cells. J. Neurochem. 44:1451–1458.

    PubMed  Google Scholar 

  8. Kanfer, J. N. 1986. The monomethylethanolamine and dimethylethanolamine base exchange reactions of a rat brain microsomal fraction. Biochim. Biophys. Acta, 879:278–285.

    PubMed  Google Scholar 

  9. Miller, E. M. 1974. Deanol: a solution for tardive dyskinesia. New England J. Med. 291:796–797.

    Google Scholar 

  10. Lewis, J. A., and Young, R. 1975. Deanol and methylphenidate in minimal brain dysfunction. Clin. Pharmacol. Ther. 17:534–540.

    PubMed  Google Scholar 

  11. Fann, W. E., Sullivan, J. L., Miller, R. D., and McKenzie, G. M. 1975. Deanol in tardive dyskinesia: a preliminary report. Psychopharmacologia 42:135–137.

    PubMed  Google Scholar 

  12. Zahniser, N. R., Chou, D. and Hanin, I. 1977. Is 2-dimethylaminoethanol (deanol) indeed a precursor of brain acetylcholine? A gas chromatographic evaluation. J. Pharmacol. Exp. Ther., 200:545–559.

    PubMed  Google Scholar 

  13. Zahniser, N. R., Katyal, S. L., Shih, T. M., Monin, I., Mossy, J., Martinez, A. J., and Lombardy, B. 1978. Effects of N-methylaminoethanol and N,N-dimethylaminoethanol in the diet of pregnant rats on neonatal rat brain cholinergic and phospholipid profile. J. Neurchem. 30:1245–1252.

    Google Scholar 

  14. Dainous, F., and Kanfer, J. N. 1986. Effect of modification of membrane phospholipid composition on phospholipid methylation in aggregating cell culture. J. Neurochem. 46:1859–1866.

    PubMed  Google Scholar 

  15. Bartlett, G. R. 1959. Phosphorus assay in column chromatography. J. Biol. Chem. 234:466–468.

    PubMed  Google Scholar 

  16. Horrocks, L. A., and Sun, G. Y. 1972. Ethanolamine plasmalogens. Research Methods in Neurochemistry. Pages 223–231,in Marks N. and Rodnight R. (eds.) Vol. 1, Raven Press, New York.

    Google Scholar 

  17. Hirata, F., Stittmatter, W. J., and Axelrod, J. 1979. β-adrenergic receptor agonists increase phospholipid methylation, membrane fuidity and β-adrenergic receptor adenylate cyclase coupling. Proc. Natl. Acad. Sci. USA 76:368–372.

    PubMed  Google Scholar 

  18. Vance, D. E., Pelech, S. D. and Choy, P. C. 1981. CTP: Phosphocholine cytidylyltransferase from rat liver. Methods in Enzymology, Pages 576–581, Enzymes of Phospholipid Synthesisin Colowick, S. P., and Kaplan, N. D., (eds.) Vol. 71, Academic Press, New York.

    Google Scholar 

  19. Fonnum, F. 1975. A rapid radiochemical method for the determination of choline acetyltransferase. J. Neurochem. 24:407–409.

    PubMed  Google Scholar 

  20. Akesson, B. 1978. Autoregulation of phospholipid-N-methylation by the membrane phosphatidylethanolamine content. F.E.B.S. Lett., 92:177–180.

    Google Scholar 

  21. Maeda, M., Tanaka, Y., and Akamatsu, Y. 1980. Phospholipid methylation in MOPC-31C cell membranes with modified phospholipid composition. Biochem. Biophys. Res. Commun. 96:876–881.

    PubMed  Google Scholar 

  22. Robert, J., Rebel, G., Mandel, P., and Yavin, E. 1977. Polar head group manipulation of phospholipids in cultured neuroblastoma cells. Life Sci. 22:211–216.

    Google Scholar 

  23. McKenzie, R. C. and Brophy, P. J. 1983. Lipid composition and physical properties of membranes from C-6 glial cells with altered phospholipid polar headgroups. Biochim. Biophys. Acta 763:357–362.

    Google Scholar 

  24. Yavin, E. 1980. Regulation of phospholipid metabolism in differentiating cells from rat brain cerebral hemispheres in culture: antogenesis of carrier specific transport of choline and N-methyl-substituted choline analogs. J. Neurochem. 34:178–183.

    PubMed  Google Scholar 

  25. Sastry, B. V., Owens, L. K., and Janson, V. E. 1982. Enhancement of the responsiveness of the rat diaphragm by L-methionine and phospholipid methylation and their relationship to aging. J. Pharmacol. Exp. Ther. 221:629–636.

    PubMed  Google Scholar 

  26. Hoffman, D. R., Haning, J. A., and Cornatzer, W. E. 1981. Effects of a methyl-deficient diet on rat liver phosphatidylcholine biosynthesis. Can. J. Biochem. 59:543–550.

    PubMed  Google Scholar 

  27. Schneider, W. J., and Vance, D. E. 1978. Effect of choline deficiency on the enzymes that synthesize phosphatidylcholine and phosphatidylethanolamine in rat liver. Europ. J. Biochem. 85:181–187.

    PubMed  Google Scholar 

  28. Ansell, G. B., and Spanner, S. 1977. Studies on the origin of choline in the brain of the rat. Biochem. J. 122:741–750.

    Google Scholar 

  29. Kanfer, J. N. 1982. Contribution of substrates for phospholipid-N-methylation by the base exchange enzyme in the rat brain microsomes. Biochem. Biophys. Res. Commun. 106:422–428.

    PubMed  Google Scholar 

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Affiliations

  1. Department of Biochemistry, Faculty of Medicine, University of Manitoba, 770 Bannatyne Avenue, R3E 0W3, Winnipeg, Canada

    Francine Dainous & Julian N. Kanfer

Authors
  1. Francine Dainous
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  2. Julian N. Kanfer
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Dainous, F., Kanfer, J.N. The incorporation of monomethylethanolamine and dimethylethanolamine in fetal brain aggregating cell culture. Neurochem Res 13, 1–8 (1988). https://doi.org/10.1007/BF00971848

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  • DOI: https://doi.org/10.1007/BF00971848

Key Words

  • Aggregating nerve cell culture
  • monomethylethanolamine
  • dimethylethanolamine
  • phospholipid-N-methyltransferase