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Essential fatty acid deficiency in freshwater fish: the effects of linoleic, α-linolenic, γ-linolenic and stearidonic acids on the metabolism of [1-14C]18:3n-3 in a carp cell culture model

Fish Physiology and Biochemistry volume 22, pages 67–75 (2000)Cite this article

Abstract

The desaturation of [1-14C]18:3n-3 to 20:5n-3 and 22:6n-3 is enhanced in an essential fatty acid deficient cell line (EPC-EFAD) in comparison with the parent cell line (EPC) from carp. In the present study, the effects of competing, unlabeled C18 polyunsaturated fatty acids (PUFA), linoleic (18:2n-6), α-linolenic (18:3n-3), γ-linolenic (18:3n-6) and stearidonic (18:4n-3) acids, on the metabolism of [1-14C]18:3n-3 were investigated in EPC-EFAD cells in comparison with EPC cells. The incorporation of [1-14C]18:3n-3 in both cell lines was significantly reduced by competing C18 PUFA, with the rank order being 18:4n-3>18:3n-3 = 18:2n-6>18:3n-6. In the absence of competing PUFA, radioactivity from [1-14C]18:3n-3 in EPC cells was predominantly recovered in phosphatidylethanolamine followed by phosphatidylcholine. This pattern was unaffected by competing n-6PUFA, but n-3PUFA reversed this pattern as did essential fatty acid deficiency in the presence of all competing PUFA. The altered lipid class distribution was most pronounced in cells supplemented with 18:4n-3. Competing C18 PUFA significantly decreased the proportions of radioactivity recovered in 22:6n-3, pentaene and tetraene products, with the proportions of radioactivity recovered in 18:3n-3 and 20:3n-3 increased, in both cell lines. However, the inhibitory effect of competing C18 PUFA on the desaturation of [1-14C]18:3n-3 was significantly greater in EPC-EFAD cells. The magnitude of the inhibitory effects of C18 PUFA on [1-14C]18:3n-3 desaturation was dependent upon the specific fatty acid with the rank order being 18:4n-3>18:3n-3>18:2n-6, with 18:3n-6 having little inhibitory effect on the metabolism of [1-14C]18:3n-3 in EPC cells. The differential effects of the C18 PUFA on [1-14C]18:3n-3 metabolism were consistent with mass competition in combination with increased desaturation activity in EPC-EFAD cells and the known substrate fatty acid specificities of desaturase enzymes. However, the mechanism underpinning the greater efficacy with which the unlabeled C18PUFA competed with [1-14C]18:3n-3 in the desaturation pathway in EPC-EFAD cells was unclear.

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References

  • Biagi, P.L., Bordoni, A., Hrelia, S., Celadon, M. and Horrobin, D.F. 1991. ?-Linolenic acid dietary supplementation can reverse the aging influence on rat liver microsome ?6-desaturase activity. Biochim. Biophys. Acta 1083: 187-192.

    Google Scholar 

  • Blond, J.P., Ratieuville, P. and Bezard, J. 1986. Acides gras alimentaires et ?5 desaturation par les microsomes hepatiques de rat. Reprod. Nutr. Devel. 26: 77-84.

    Google Scholar 

  • Brenner, R.R. 1981. Nutritional and hormonal factors influencing desaturation of essential fatty acids. Prog. Lipid Res. 20: 41-47.

    Google Scholar 

  • Capriotti, A.M. and Laposta, M. 1986. Identification of variables critical to reproducible delipidation of serum. J. Tissue Culture Methods 10: 219-221.

    Google Scholar 

  • Choi, Y.S. and Sugano, M. 1988. Effects of dietary ?-and ?-linolenic acids on lipid metabolism in young and adult rats. Ann. Nutr. Metab. 32: 169-176.

    Google Scholar 

  • Christie, W.W. 1982. Lipid Analysis, 2nd edn. Pergamon Press, Oxford.

    Google Scholar 

  • Folch, J., Lees, M., Sloane Stanley, G.H. 1957. A simple method for the isolation and purification of total lipids from animal tissues. J. Biol. Chem. 226: 497-509.

    Google Scholar 

  • Ghioni, C., Tocher, D.R. and Sargent, J.R. 1997. The effect of culture on morphology, lipid and fatty acid composition, and polyunsaturated fatty acid metabolism of rainbow trout (Oncorhynchus mykiss) skin cells. Fish Physiol. Biochem. 16: 499-513.

    Google Scholar 

  • Hansen, H.S. 1986. The essential nature of linoleic acid in mammals. Trends Biochem. Sci. 11: 263-265.

    Google Scholar 

  • Hoy, C.E., Holmer, G., Kaur, N., Byrjalsen, I. and Kirstein, D. 1983. Acyl group distributions in tissue lipids of rats fed evening primrose oil (?-linolenic plus linoleic acid) or soybean oil (?-linolenic plus linoleic acid). Lipids 18: 760-771.

    Google Scholar 

  • Laposata, M., Prescott, S.M., Bross, T.E. and Majerus, P.W. 1982. Development and characterization of a tissue culture cell line with essential fatty acid deficiency. Proc. Natl. Acad. Sci. USA 79: 7654-7658.

    Google Scholar 

  • Lerner, R., Lindstrom, P., Berg, A., Johansson, E., Rosendahl, K. and Palmblad, J. 1995. Development and characterization of essential fatty acid deficiency in human endothelial cells in culture. Proc. Natl. Acad. Sci. USA 92: 1147-1151.

    Google Scholar 

  • Marcelo, C.L., Duell, E.A., Rhodes, L.M. and Dunham, W.R. 1992. In vitro model of essential fatty acid deficiency. J. Invest. Dermatol. 99: 703-708.

    Google Scholar 

  • Melin, T. and Nilsson, A. (1997) ?6 desaturase and ?5 desaturase in human Hep G2 cells are both fatty acid interconversion rate limiting and are upregulated under essential fatty acid deficient conditions. Prostaglandins Leukotrienes Essent. Fatty Acids 56: 437-442.

    Google Scholar 

  • Sargent, J.R., Bell, J.G., Bell, M.V., Henderson, R.J. and Tocher, D.R. 1995. Requirement criteria for essential fatty acids. J. Appl. Icthyol. 11: 183-198.

    Google Scholar 

  • Takeuchi, T. and Watanabe, T. 1977. Requirement of carp for essential fatty acids. Bull. Jap. Soc. Sci. Fish. 43: 541-551.

    Google Scholar 

  • Tocher, D.R. and Dick, J.R. 1999. Polyunsaturated fatty acid metabolism in a cell culture model of essential fatty acid defi-ciency in a freshwater fish, carp (Cyprinus carpio). Fish Physiol. Biochem., 21: 257-267.

    Google Scholar 

  • Tocher, D.R., Bell, J.G., Dick, J.R. and Sargent, J.R. 1997. Fatty acyl desaturation in isolated hepatocytes from Atlantic salmon (Salmo salar): Stimulation by dietary borage oil containing-linolenic acid. Lipids 32: 1237-1247.

    Google Scholar 

  • Tocher, D.R. Dick, J.R. and Sargent, J.R. 1995. Development of an in vitro model of essential fatty acid deficiency in fish cells. Prostaglandins Leukotrienes Essent. Fatty Acids 53: 365-375.

    Google Scholar 

  • Tocher, D.R., Dick, J.R. and Sargent, J.R. 1996. Stimulation of proliferation of an essential fatty acid deficient fish cell line by C20 and C22 polyunsaturated fatty acids and effects on fatty acid composition. Prostaglandins Leukotrienes Essent. Fatty Acids 55: 345-356.

    Google Scholar 

  • Tocher, D.R., Sargent, J.R. and Frerichs, G.N. 1988. The fatty acid compositions of established fish cell lines after long-term culture in mammalian sera. Fish Physiol. Biochem. 5: 219-227.

    Google Scholar 

  • Ulmann, L., Blond, J.P., Maniongui, C., Poisson, J.P., Durand, G., Bezard, J. and Pascal, G. 1991. Effects of age and dietary fatty acids on desaturase activities and on fatty acid composition of liver microsomal phospholipids of adult rats. Lipids 26: 127-133.

    Google Scholar 

  • Vitiello, F., and Zanetta, J.-P. 1978. Thin-layer chromatography of phospholipids. J. Chromatogr. 166: 637-640.

    Google Scholar 

  • Wilson, R. and Sargent, J.R. 1992. High resolution separation of polyunsaturated fatty acids by argentation thin-layer chromatography. J. Chromatogr. 623: 403-407.

    Google Scholar 

  • Zar, J.H. 1984. Biostatistical Analysis, 2nd edition. Prentice-Hall, Englewood Cliffs.

    Google Scholar 

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Authors and Affiliations

  1. Unit of Aquatic Biochemistry, Institute of Aquaculture, University of Stirling, Stirling , FK9 4LA, Scotland, UK

    Douglas R. Tocher & James R. Dick

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  1. Douglas R. Tocher
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  2. James R. Dick
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Tocher, D.R., Dick, J.R. Essential fatty acid deficiency in freshwater fish: the effects of linoleic, α-linolenic, γ-linolenic and stearidonic acids on the metabolism of [1-14C]18:3n-3 in a carp cell culture model. Fish Physiology and Biochemistry 22, 67–75 (2000). https://doi.org/10.1023/A:1007877130756

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  • DOI: https://doi.org/10.1023/A:1007877130756

  • carp
  • cell culture
  • desaturation
  • elongation
  • essential fatty acid deficiency
  • polyunsaturated fatty acids

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