Association and Expression of CYP2A6 and KIF12 Genes Related to Lamb Flavour and Odour
AbstractCytochrome P450, family 2, subfamily A, polypeptide 6 (CYP2A6) and kinesin-like protein KIF12 (KIF12) genes are predicted as candidate genes which play important roles in lamb flavour and odour. The aim of this study was to analyse the genotype polymorphism of CYP2A6 and KIF12 genes, to study association and expression of these genes with lamb flavour and odour. Identification of genes polymorphism and associations of CYP2A6 and KIF12 genes were performed using PCR-RFLP method and GLM analysis. The PCR-RFLP products of CYP2A6 and KIF12 were digested by restriction enzyme BsmAI and BfaI, respectively. The expression of CYP2A6 gene was performed using qRT-PCR. The results showed that the CYP2A6 and KIF12 genes were polymorphics. The CYP2A6 gene found to have two genotypes (TT and GT), whereas the KIF12 gene found to have three genotypes (CC, CT, and TT). The CYP2A6 and KIF12 genes were in Hardy Weinberg Equilibrium (HWE). Association analysis showed that CYP2A6 (g.49170107 G>T) was significantly (P<0.05) associated with 3-methylindole (MI) or skatole, while KIF12 (g.9617965 C>T) was not significantly associated with lamb flavour and odour. The GT genotype exhibited a greater 3-methylindole (MI) or skatole than the TT genotype (P<0.05). The mRNA expression analysis showed that CYP2A6 mRNA expression was higher (P<0.01) in animals with the TT genotype. These results will improve the understanding of the functions of the CYP2A6 in lamb flavour and odour, especially in term of 3-methylindole (MI) or skatole compound within the liver and will shed light on CYP2A6 as a candidate in the selection of sheep with low lamb flavour and odour.
Albertí, P., B. Panea, C. Sañudo, J. L. Olleta, G. Ripoll, P. Ertbjerg, M. Christensen, S. Gigli, S. Failla, S. Concetti, J. F. Hocquette, R. Jailler, S. Rudel, G. Renand, G. R. Nute, R. I. Richardson, & J. L. Williams. 2008. Live weight, body size and carcass characteristics of young bulls of fifteen european breeds. Livestock Sci. 114:19-30. https://doi.org/10.1016/j.livsci.2007.04.010
Allendorf, F. W., G. Luikart, & S. N. Aitken. 2013. Conservation and the genetics of populations. John Wiley & Sons, Chichester.
Awan, K., S. A. Khan, M. M. Khan, & M. T. Khan. 2014. Effect of age on physico-chemical and sensorial quality of buffalo meat. Glob Vet. 13:28-32.
Bain, A., D. A. Astuti, S. Suharti, C. Arman, & K. G. Wiryawan. 2016. Performance, nutrient digestibility, and meat quality of bali cattle fed a ration supplemented with soybean oil calcium soap and cashew fruit flour. Med. Pet. 39:180-188. https://doi.org/10.5398/medpet.2016.39.3.180
Blanch, M., N. Panella-Riera, P. Chevillon, M. Font i Furnols, M. Gil, J. M. Gil, & et al. 2012. Impact on consumer’s sensitivity to androstenone on acceptability of meat grom entire male pigs in three European countries: France, Spain and United Kingdom. Meat Sci. 90:572-578. https://doi.org/10.1016/j.meatsci.2011.09.018
Bonneau, M., & Chevillon, P. 2012. Acceptability of entire male pork with various levels of androstenone and skatole by consumers according to their sensitivity to androstenone. Meat Sci. 90:330-337. https://doi.org/10.1016/j.meatsci.2011.07.019
Bourdon, R. M. 2000. Understanding Animal Breeding. 2nd Ed. Prentice Hall Inc. Upper Saddle River, New Jersey.
Chen, G., Cue, R. A., Lundstrom, K., Wwood, J. D., & Doran, O. 2008. Regulation of CYP2A6 protein expression by skatole, indole, and testicular steroids in primary cultured pig hepatocytes. Drug Metab Dispos. 36:56-60. https://doi.org/10.1124/dmd.107.017285
Cinar, M. U., Kayan, A., Uddin, M. J., Jonas, E., Tesfaye, D., Phatsara, C., Ponsuksili, S., Wimmers, K., Tholen, E., Looft, C., Jungst, H., & Schellander, K. 2012. Association and expression quantitative trait loci (eQTL) analysis of porcine AMBP, GC and PPP1R3B genes with meat quality traits. Mol Biol Rep. 39:4809-4821. https://doi.org/10.1007/s11033-011-1274-4
Diaz, G. J., & E. J Squires. 2000. Metabolism of 3-methylindole by porcine liver microsomes: responsible cytochrome P450 enzymes. Toxicol Sci. 55:284-292. https://doi.org/10.1093/toxsci/55.2.284
Duijvesteijn, N., Knol, E. F., Merks, J. W. M., Crooijmans, R., Groenen, M. A. M., Bovenhuis, H., & Harlizius, B. 2010. A genome-wide association study on androstenone levels in pigs reveals a cluster of candidate genes on chromosome 6. BMC Genet. 11:42. https://doi.org/10.1186/1471-2156-11-42
Fan, B., DU, Z. Q., & Rothschild, M. F. 2010. The hepatocyte nuclear factor-1 alpha (HNF1A) gene is associated with fatness and loin muscle area in the pig. Animal. 4:1619-1627. https://doi.org/10.1017/S175173111000087X
Grindflek, E., Lien, S., Hamland, H., Hansen, MH., Kent, M., & et al. 2011. Large scale genome-wide association and LDLA mapping study identifies QTLs for boar taint and related sex steroids. BMC Genomics. 12: 362. https://doi.org/10.1186/1471-2164-12-362
Gunawan, A., Kaewmala, K., Uddin, M. J., Cinar, M. U., Tesfaye, D., Phatsara, C., & et al. 2011. Association study and expression analysis of porcine ESR1 as a candidate gene for boar fertility and sperm quality. Anim Reprod Sci. 128:11-21. https://doi.org/10.1016/j.anireprosci.2011.08.008
Gunawan, A., Sahadevan, S., Cinar, M. U., Neuhoff, C., Große-Brinkhaus, C., Frieden, L., Tesfaye, D., Tholen, E., Looft, C., Wondim, D. S., & Hölker, M. 2013a. Identification of the novel candidate genes and variants in boar liver tissues with divergent skatole levels using RNA deep sequencing. PLoS One. 8(8).p.e72298. https://doi.org/10.1371/journal.pone.0072298
Gunawan, A., Sahadevan, S., Neuhoff, C., Große-Brinkhaus, C., Gad, A., Frieden, L., Tesfaye, D., Tholen, E., Looft, C., Uddin, M. J., & Schellander, K. 2013b. RNA deep sequencing reveals novel candidate genes and polymorphisms in boar testis and liver tissues with divergent androstenone levels. PloS one. 8(5), p.e63259. https://doi.org/10.1371/journal.pone.0063259
Gunawan, A., Jakaria, Listyarini, K., Furqon, A., Sumantri, C., Akter S. H., & Uddin M. J. 2018. Transcriptome signature of liver tissue with divergent sheepmeat odour and flavour using RNA deep sequencing. Gene. 676:86-94.
Hartl, D. L., & Clark, A. G. 1997. Principle of Population Genetic. Sinauer Associates, Sunderland, MA.
Henseler, S., Preuss, S., & Bennewitz, J. 2014. Fleischerzeugung mitMerinolandschaf-Gebrauchskreuzungen – 2. Mitteilung, sensorische Analyse und Zusammenführung der Ergebnisse in einem Index. Züchtungskunde. 86:104-115.
Hirokawa, N., Noda, Y., Tanaka, Y., & Niwa, S. 2009. Kinesin superfamily motor proteins and intracellular transport. Nat Rev Mol Cell Biol. 10:682-696. https://doi.org/10.1038/nrm2774
Hoffman, T. W., Belk, K. E., Woerner, D. R., Tatum, J. D., Delmore, R. J., Peel, R. K., LeValley, S. B., Pendell, D. L., Maneotis, K. A., Zerby, H. N., English, L. F., Moeller, S. J., & Fluharty, F. L. 2015. Executive Summary Preferences and Complaints associated with American Lamb Quality in Retail & Foodservice Markets. Am. Lamb Board. Denver, CO.
Hopkins, D. L., Fogarty, N. M., & Mortimer, S. I. 2011. Genetic related effects on sheep meat quality. Small Ruminant Res. 101:160-72. https://doi.org/10.1016/j.smallrumres.2011.09.036
Jayasena, D. D., Jung, S., Kim, H. J., Alahakoon, A. U., Nam, K. C., & Jo, C. 2014. Effect of Sex on Flavor-related and Functional Compounds in Freeze-dried Broth Made from Korean Native Chicken. Korean J Food Sci Anim Resour. 34:448-456. https://doi.org/10.5851/kosfa.2014.34.4.448
Kayan, A., Cinar, M. U., Uddin, M. J., Phatsara, C., Wimmers, K., Ponsuksili, S., Tesfaye, D., Looft, C., Juengst, H., Tholen, E., & Schellander, K. 2011. Polymorphism and expression of the porcine Tenascin C gene associated with meat and carcass quality. Meat Sci. 89:76-83. https://doi.org/10.1016/j.meatsci.2011.04.001
Lanthier, F., Lou, Y., & Squires, E. J. 2007. Skatole metabolism in the intact pre-pubescent male pig: The relationship between hepatic enzyme activity and skatole concentrations in plasma and fat. Livest Sci. 106:145-153. https://doi.org/10.1016/j.livsci.2006.07.009
Lee, G. J., Archibald, A. L., Law, A. S., Lloyd, S., Wood, J., & Haley, C. S. 2005. Detection of quantitative trait loci for androstenone, skatole and boar taint in a cross between Large White and Meishan pigs. Anim Genet. 36:14-22. https://doi.org/10.1111/j.1365-2052.2004.01214.x
Lin, Z., Lou, Y., & Squires, E. J. 2004. Molecular cloning, expression and functional characterisation of the cytochrome P450 2A6 gene in pig liver. Anim Genet. 35:314-316. https://doi.org/10.1111/j.1365-2052.2004.01140.x
Matal, J., Mayuskova, Z., Tunkova, A., Anzenbacherova, E., & Anzenbacher, P. 2009. Porcine CYP2A19, CYP2E1, and CYP1A2 Forms are Responsible for Skatole Biotransformation in the Reconstituted System. Neuroendocrinol Lett. 30:36-40.
McRae, J. M., Jaeger, S. R., Bava, C. M., Beresford, M. K., Hunter, D., Jia, Y., Chheang, S. L., Jin, D., Peng, M., Gamble, J. C., Atkinson, K. R., Axten, L. G., Paisley, A. G., William, L., Tooman, L., Pineau, B., Rouse, S. A., & Newcomb, R. D. 2013. Identification of regions associated with variation in sensitivy to food related odors in the human genome. Curr Biol. 23:1596-1600. https://doi.org/10.1016/j.cub.2013.07.031
Moe, M., Lien, S., Aasmundstad, T., Meuwissen, T. H. E., Hansen, M. H. S., Bendixen, C., & Grindflek, E. 2009. Association between SNPs within candidate genes and compounds related to boar taint and reproduction. BMC Genet. 10:32. https://doi.org/10.1186/1471-2156-10-32
Morlein, D., Lungershausen, M., Steinke, K., Sharifi, A. R., & Knorr, C. 2012. A single nucleotide polymorphism in the CYP2E1 gene promoter affects skatole content in backfat of boars of two commercial Duroc-sired crossbred populations. Meat Sci. 92:739-744. https://doi.org/10.1016/j.meatsci.2012.06.031
Nei, & Kumar. 2000. Moleculear Evolution and Phylogenetics. Oxford University Press, New York.
Neuhoff, C., Gunawan, A., Farooq, M. O., Cinar, M. U., Brinkhaus, C. G., Sahadevan, S., Frieden, L., Tesfaye, D., Tholen, E., Looft, C., & et al. 2015. Preliminary study of FMO1, FMO5, CYP21, ESR1, PLIN2, and SULT2A1 as candidate gene for compounds related to boar taint. Meat Sci. 108:67-73. https://doi.org/10.1016/j.meatsci.2015.05.025
Ran-Ressler R. R., Bae, S., Lawrence, P., Wang, D. H., & Brenna J. T. 2014. Branched-chain fatty acid content of foods and estimated intake in the USA. Br J Nutr. 112:565-572. https://doi.org/10.1017/S0007114514001081
Robic, A., Feve, K., Larzul, C., Billon, Y., & Van Son, M. 2011. Expression levels of 25 genes in liver and testis located in a QTL region for androstenone on SSC7q1.2. Anim Genet. 42:662-665. https://doi.org/10.1111/j.1365-2052.2011.02195.x
Sambrook, J., & Russell, D. 2001. Molecular Cloning: a Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press, New York.
Schiller, K. F., Preuss, S., Kaffarnik, S., Vetter, W., Rodehutscord, M., & Bennewitz, J. 2015. Concentration of three branched-chain fatty acids in adipose tissue does not affect meat sensory traits in crossbred and purebred German “Merinolandschaf” lambs. Arch Anim Breed. 58:159-163. https://doi.org/10.5194/aab-58-159-2015
Silver, N., Best, S., Jiang, J., & Thein, S. L. 2006. Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR. BMC Mol Biol. 7:33. https://doi.org/10.1186/1471-2199-7-33
Strathe, A. B., Velander, I. H., Mark, T., & Kadarmideen, H. N. 2013. Genetic parameters for androstenone and skatole as indicators of boar taint and their relationship to production and litter size traits in danish landrace. Anim Sci J. 91:2587-2595. https://doi.org/10.2527/jas.2012-6107
Sumantri, C., R. Diyono, A. Farajallah & I. Inounu. 2008. Polymorphism of calpastatin gene and its eﬀect on body weight of local sheeps. JITV. 13:117-126.
Tajeda, J. F., Pena, R. E., & Andres, A. I. 2008. Effect of live weight and sex on physico-chemical and sensorial characteristics of Merino lamb meat. Meat Sci. 80:1061-1067. https://doi.org/10.1016/j.meatsci.2008.04.026
Ueno, H., Huang, X., Tanaka, Y., & Hirokawa, N. 2011. KIF16B/Rab14 molecular motor complex is critical for early embryonic development by transporting FGF receptor. Dev Cell. 20:60-71. https://doi.org/10.1016/j.devcel.2010.11.008
Watkins, P. J., Rose, G., Salvatore, L., Allen, D., Tucman, D., Warner, R. D., Dunshea, F. R., & Pethick, D. W. 2010. Age and nutrition influence the concentrations of three branched chain fatty acids in sheep fat from Australian abattoirs. Meat Sci. 86:594-599. https://doi.org/10.1016/j.meatsci.2010.04.009
Watkins, P. J., Kearney, G., Rose, G., Allen, D., Ball, A. J. D., Pethick W., & Warner, R. D. 2014. Effect of branched-chain fatty acids 3-methylindole and 4-methylphenol on consumer sensory scores of grilled lamb meat. Meat Sci. 96:1088-1094. https://doi.org/10.1016/j.meatsci.2012.08.011
Whittington, F. M., Zammerini, D., Nute, G. R., Baker, A., Hughes, S. I., & Wood, J. D. 2011. Comparison of heating methods and the use of different tissues for sensory assessment of abnormal odours (boar taint) in pig meat. Meat Sci. 88:249-255. https://doi.org/10.1016/j.meatsci.2010.12.029
Wiercinska, P., Lou, Y., & Squires, E. J. 2012. The role of different procine cytochrome P450 enzymes and cytochrome b5A in skatole metabolism. SAPT. 6: 834-845.
Windig, J. J., Mulder, H. A., Ten Napel, J., Knol, E. F., Mathur, P. K., & Crump, R. E. 2012. Genetic parameters for androstenone, skatole, indole and human nose scores as measures ofboar taint and their relationship with finishing traits. Anim Sci J. 90:2120-2129. https://doi.org/10.2527/jas.2011-4700
Wood, J., Enser, M., Fisher, A., Nute, G., Sheard, P., Richardson, R. & et al. 2008. Fat deposition, fatty acid composition and meat quality: A review. Meat Sci. 78:343-353. https://doi.org/10.1016/j.meatsci.2007.07.019
Yang, W., Tanaka, Y., Bundo, M., & Hirokawa, N. 2014. Antioksidant signaling involving the microtubule motor KIF12 is an intracellular target of nutrition excess in beta cells. Dev Cell. 31:202-214. https://doi.org/10.1016/j.devcel.2014.08.028
Young, O. A., Lane, G. A., Priolo, A., & Fraser, K. 2003. Pastoral and species flavour in lambs raised on pasture, lucerne or maize. J. Sci. Food Agric. 83:93-104. https://doi.org/10.1002/jsfa.1282
Zamaratskaia, G., & Squires, E. J. 2009. Biochemical, nutritional and genetic effects on boar taint in entire male pigs. Animal. 3:1508-1521. https://doi.org/10.1017/S1751731108003674
Zhou, R., Niwa, S., Homma, N., Takei, Y., & Hirokawa, N. 2009. KIF26A is an unconventional kinesin and regulates GDNF-Ret signaling in enteric neuronal development. Cell. 139:802-813. https://doi.org/10.1016/j.cell.2009.10.023
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