Hostname: page-component-586b7cd67f-vdxz6 Total loading time: 0 Render date: 2024-11-30T23:53:46.309Z Has data issue: false hasContentIssue false

The relationship between adipose tissue skatole levels, rates of hepatic microsomal skatole metabolism and hepatic cytochrome P450IIE1 expression in two breeds of pig

Published online by Cambridge University Press:  18 August 2016

E. Doran
Affiliation:
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK
F. W. Whittington
Affiliation:
Department of Clinical Veterinary Science, School of Veterinary Science, Langford, University of Bristol, Bristol BS40 5DU, UK
J. D. Wood
Affiliation:
Department of Clinical Veterinary Science, School of Veterinary Science, Langford, University of Bristol, Bristol BS40 5DU, UK
J. D. McGivan*
Affiliation:
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK
*
Corresponding author; E-mail:[email protected]
Get access

Abstract

The relationships between the level of skatole in backfat, the rate of skatole metabolism in isolated liver microsomes, hepatic cytochrome P450IIE1 content and mRNA levels were investigated in Large White ✕ Landrace (LW) and the Meishan ✕ Landrace (M) breeds. A method based on thin layer chromatography was developed and used for measurement of microsomal skatole metabolism. Skatole metabolism by liver microsomes was inhibited by diallyl sulphide, a specific inhibitor of cytochrome P450IIE1 but not by inhibitors of other P450 isoforms. We have shown that the rate of skatole metabolism by liver microsomes was proportional to the microsomal P450IIE1 content. In LW pigs there was considerable variation in cytochrome P450IIE1 expression and P450IIE1 protein level and there was a significant negative correlation between backfat skatole level and hepatic microsomal cytochrome P450IIE1 content. Pigs exhibiting low P450IIE1 content in general also showed low levels of P450IIE1 mRNA. These results show that the levels of deposition of backfat skatole in LW pigs are inversely related to the rate at which skatole can be metabolized by liver microsomes, and this in turn depends on the level of expression of cytochrome P450IIE1 mRNA in the liver. In the M breed generally the P450IIE1 protein and mRNA levels were very low and backfat skatole level was high. However the skatole level varied over a 10-fold range and there was no significant correlation with P450IIE1 protein or mRNA content. The M breed was shown to express high levels of the multidrug resistance protein in liver, and it is suggested that export of skatole from liver via this transport protein may be an additional factor regulating backfat skatole in M pigs, but not in the LW breed.

Type
Growth, development and meat science
Copyright
Copyright © British Society of Animal Science 2002

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Annor-Frempong, I.E., Nute, G. R., Whittington, F. W. and Wood, J. D. 1997. The problem of boar taint in pork. II. The influence of androstenone, skatole and indole, presented individually and in combination, on odour perception. Meat Science 47: 4961.Google Scholar
Babol, J., Squires, E. J. and Lundstrom, K. 1998a. Hepatic metabolism of skatole in pigs by cytochrome P4502E1. Journal of Animal Science 76: 822828.Google Scholar
Babol, J., Squires, E. J. and Lundstrom, K. 1998b. Relationship between oxidation and conjugation metabolism of skatole in pig liver and concentration of skatole in fat. Journal of Animal Science 76: 829838.Google Scholar
Banerjee, S., Ganapathi, R., Ghosh, L. and Yu, C. L. 1992. Down-regulation of ras and myc expression associated with mdr-1 overexpression in adriamycin-resistant tumor cells. Cellular and Molecular Biology 38: 561570.Google Scholar
Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248254.Google Scholar
Cleveland, D. W., Fischer, S. G., Kirschner, M. W. and Laemmli, U. K. 1977. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. Journal of Biological Chemistry 252: 11021106.Google Scholar
Diaz, G. J. and Squires, E. J. 2000. Metabolism of 3-methylindole by porcine liver microsomes: responsible cytochrome P450 enzymes. Toxicological Sciences 55: 284292.Google Scholar
Gottesman, M. M. and Pastan, I. 1993. Biochemistry of multidrug resistance mediated by the multidrug transporter (review). Annual Review of Biochemistry 62: 385427.Google Scholar
Gupta, S. and Gollapudi, S. 1993. P-glycoprotein (MDR 1 gene product) in cells of the immune system: its possible physiologic role and alteration in aging and human immunodeficiency virus-1 (HIV-1) infection. Journal of Clinical Immunology 13: 289301.Google Scholar
Lundstrom, K. and Bonneau, M. 1996. Off flavour of meat with particular emphasis on boar taint. In ECCEAMST (ed. Taylor, S. Raimundo, A. Severini, M. and F. Smulders, J. M.), pp. 137154. Utrecht, The Netherlands.Google Scholar
Porter, T. D., Khani, S. C. and Coon, M. J. 1989. Induction and tissue-specific expression of rabbit cytochrome P4502E1 and 2E2 genes. Molecular Pharmacology 36: 6165.Google Scholar
Sambrook, J., Fritsch, E. F. and Maniatis, T. 1989. Analysis of RNA. In Molecular cloning. A laboratory manual (ed. Nolan, C.), pp. 7·37–7·57. Cold Spring Harbor, New York.Google Scholar
Schenkman, J. B. and Cinti, D. L. 1978. Preparation of microsomes with calcium. Methods in Enzymology 52: 8389.Google Scholar
Squires, E. J. and Lundstrom, K. 1997. Relationship between cytochrome P4502E1 in liver and levels of skatole and its metabolites in intact male pigs. Journal of Animal Science 75: 25062511.Google Scholar
Wiseman, J., Redshaw, M. S., Jagger, S., Nute, G. R., Whittington, F. W. and Wood, J. D. 1999. Influence of type and dietary rate of inclusion of non-starch polysaccharides on skatole content and meat quality of finishing pigs. Animal Science 69: 123133.Google Scholar
Zaman, G. J., Flens, M. J., Leusden, M. R.van, Haas, M.de, Mulder, H. S., Lankelma, J., Pinedo, H. M., Sheper, R. J., Baas, F., Broxterman, H. J. and Borst, P. 1994. The human multidrug resistance-associated protein MRP is a plasma membrane drug-efflux pump. Proceedings of the National Academy of Sciences of the United States of America 91: 88228826.Google Scholar