mRNA Expression of CYP2E1, CYP2A19, CYP1A2, HSD3B, SULT1A1 and SULT2A1 genes in surgically castrated, immunologically castrated, entire male and female pigs and correlation with androstenone, skatole, indole and Improvac-specific antibody levels

https://doi.org/10.17221/159/2018-CJASCitation:Kubešová A., Šťastný K., Faldyna M., Sládek Z., Steinhauserová I., Bořilová G., Knoll A. (2019): mRNA Expression of CYP2E1, CYP2A19, CYP1A2, HSD3B, SULT1A1 and SULT2A1 genes in surgically castrated, immunologically castrated, entire male and female pigs and correlation with androstenone, skatole, indole and Improvac-specific antibody levels. Czech J. Anim. Sci., 64: 89-97.
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This study aimed to obtain a comprehensive look at the influence of castration on mRNA expression of the genes CYP2E1, CYP1A2, CYP2A19, HSD3B, SULT2A1 and SULT1A1 and their correlation with boar taint compounds (androstenone, skatole and indole) and Improvac-specific antibodies in a Czech commercial hybrid (Large White × Landrace (sow) × Duroc (boar)). Pigs were divided into groups of entire male pigs (NC), pigs castrated surgically (SC), pigs immunologically castrated and slaughtered 8 weeks (IM8) or 15 weeks (IM15) after the second dose of Improvac, and gilts (GI). Hepatic mRNA expression, measured by quantitative real-time polymerase chain reaction, differed significantly between the control group (entire male pigs) and all groups of interest for CYP2E1, CYP1A2 and CYP2A19. The mRNA level of the HSD3B gene differed significantly between the control group and the IM8, IM15 and GI groups. SULT1A1 gene expression was significantly different between the control group and the SC, IM8 and GI. In the case of SULT2A1, a significant difference was observed only between the control group and IM8 pigs. For all genes and treatment groups described above, expression was increased relative to the control. Significant differences for Improvac-specific antibodies between IM8 and IM15 groups were observed, indicating decrease of antibodies over time. Moreover, negative correlations between androstenone and mRNA levels of CYP2A19, CYP2E1 and SULT1A1 suggest that gene expression is suppressed.

References:
Babol J, Squires E J, Lundström K (1999): Relationship between metabolism of androstenone and skatole in intact male pigs.. Journal of Animal Science, 77, 84-  https://doi.org/10.2527/1999.77184x
 
Bidanel J., Riquet J., Gruand J., Squires E., Bonneau M., Milan D. (2006): Detection of quantitative trait loci for skatole and indole levels in Meishan × Large White F2 pigs. In: Proc. 8th World Congress on Genetics Applied to Livestock Production, Belo Horizonte, Brazil.
 
Brunius C., Rasmussen M. K., Lacoutière H., Andersson K., Ekstrand B., Zamaratskaia G. (2012): Expression and activities of hepatic cytochrome P450 (CYP1A, CYP2A and CYP2E1) in entire and castrated male pigs. animal, 6, 271-277  https://doi.org/10.1017/S1751731111001674
 
Chen G., Bourneuf E., Marklund S., Zamaratskaia G., Madej A., Lundström K. (2007): Gene expression of 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase in relation to androstenone, testosterone, and estrone sulphate in gonadally intact male and castrated pigs1. Journal of Animal Science, 85, 2457-2463  https://doi.org/10.2527/jas.2007-0087
 
Claus Rolf, Rottner Silke, Rueckert Christine (2008): Individual return to Leydig cell function after GnRH-immunization of boars. Vaccine, 26, 4571-4578  https://doi.org/10.1016/j.vaccine.2008.05.085
 
Doran Elena, Whittington Frances W., Wood Jeffrey D., McGivan John D. (2002): Cytochrome P450IIE1 (CYP2E1) is induced by skatole and this induction is blocked by androstenone in isolated pig hepatocytes. Chemico-Biological Interactions, 140, 81-92  https://doi.org/10.1016/S0009-2797(02)00015-7
 
Doran Elena, Whittington Frances M, Wood Jeffrey D, McGivan John D (2004): Characterisation of androstenone metabolism in pig liver microsomes. Chemico-Biological Interactions, 147, 141-149  https://doi.org/10.1016/j.cbi.2003.12.002
 
Duijvesteijn Naomi, Knol Egbert F, Merks Jan WM, Crooijmans Richard PMA, Groenen Martien AM, Bovenhuis Henk, Harlizius Barbara (2010): A genome-wide association study on androstenone levels in pigs reveals a cluster of candidate genes on chromosome 6. BMC Genetics, 11, -  https://doi.org/10.1186/1471-2156-11-42
 
Gillberg Mette, Skaanild Mette T., Friis Christian (2006): Regulation of Gender-Dependent CYP2A Expression in Pigs: Involvement of Androgens and CAR. Basic & Clinical Pharmacology & Toxicology, 98, 480-487  https://doi.org/10.1111/j.1742-7843.2006.pto_261.x
 
Kluivers-Poodt M., Houx B. B., Robben S. R. M., Koop G., Lambooij E., Hellebrekers L. J. (2012): Effects of a local anaesthetic and NSAID in castration of piglets, on the acute pain responses, growth and mortality. animal, 6, 1469-1475  https://doi.org/10.1017/S1751731112000547
 
Kojima Misaki, Degawa Masakuni (2013): Serum androgen level is determined by autosomal dominant inheritance and regulates sex-related CYP genes in pigs. Biochemical and Biophysical Research Communications, 430, 833-838  https://doi.org/10.1016/j.bbrc.2012.11.060
 
Kojima Misaki, Sekimoto Masashi, Degawa Masakuni (2010): Androgen-mediated down-regulation of CYP1A subfamily genes in the pig liver. Journal of Endocrinology, 207, 203-211  https://doi.org/10.1677/JOE-10-0160
 
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. Livestock Science, 106, 145-153  https://doi.org/10.1016/j.livsci.2006.07.009
 
Lin Zhihong, Lou Yanping, Squires James E. (2004): Molecular cloning and functional analysis of porcine SULT1A1 gene and its variant: a single mutation SULT1A1 causes a significant decrease in sulfation activity. Mammalian Genome, 15, 218-226  https://doi.org/10.1007/s00335-002-2318-4
 
Matal J., Matuskova Z., Tunkova A., Anzenbacherova E., Anzenbacher P. (2009): Porcine CYP2A19, CYP2E1 and CYP1A2 forms are responsible for skatole biotrasformation in the reconstituted system. Neuroendocrinology Letters, 30, 36–40.
 
Moe M., Grindflek E., Doran O. (2007): Expression of 3β-hydroxysteroid dehydrogenase, cytochrome P450-c17, and sulfotransferase 2B1 proteins in liver and testis of pigs of two breeds: Relationship with adipose tissue androstenone concentration1. Journal of Animal Science, 85, 2924-2931  https://doi.org/10.2527/jas.2007-0283
 
Nicolau-Solano S. I., McGivan J. D., Whittington F. M., Nieuwhof G. J., Wood J. D., Doran O. (2006): Relationship between the expression of hepatic but not testicular 3β-hydroxysteroid dehydrogenase with androstenone deposition in pig adipose tissue1. Journal of Animal Science, 84, 2809-2817  https://doi.org/10.2527/jas.2005-595
 
Nygard A.B., Jorgensen C.B., Cirera S., Fredholm M. (2007): Selection of reference genes for gene expression studies in pig tissue using SYBR green qPCR. BMC Molecular Biology, 8, 67. Available at: http://www.biomedcentral.com/1471-2199/8/67 (accessed Dec 9, 2016).
 
Prunier A., Bonneau M., von Borell E.H., Cinott S., Gunn M., Fredriksen B., Giersing M., Morton D.B., Tuyttens F.A.M., Velarde A. (2006): A review of the welfare consequences of surgical castration in piglets and the evaluation of non-surgical methods. Animal Welfare, 15, 277–289.
 
Rasmussen MK, Zamaratskaia G, Ekstrand B (2011): Gender-related Differences in Cytochrome P450 in Porcine Liver - Implication for Activity, Expression and Inhibition by Testicular Steroids. Reproduction in Domestic Animals, 46, 616-623  https://doi.org/10.1111/j.1439-0531.2010.1714.x
 
Rasmussen Martin Krøyer, Brunius Carl, Ekstrand Bo, Zamaratskaia Galia (2012): Expression of hepatic 3β-hydroxysteroid dehydrogenase and sulfotransferase 2A1 in entire and castrated male pigs. Molecular Biology Reports, 39, 7927-7932  https://doi.org/10.1007/s11033-012-1637-5
 
Sinclair P A, Gilmore W J, Lin Z, Lou Y, Squires E J (2006): Molecular cloning and regulation of porcine SULT2A1: relationship between SULT2A1 expression and sulfoconjugation of androstenone. Journal of Molecular Endocrinology, 36, 301-311  https://doi.org/10.1677/jme.1.01847
 
Sladek Z., Prudikova M., Knoll A., Kulich P., Steinhauserova I., Borilova G. (2018): Effect of early immunocastration on testicular histology in pigs. Veterinární Medicína, 63, 18-27  https://doi.org/10.17221/121/2017-VETMED
 
Svobodova K. (2011): Analysis of variability and expression of genes for eukaryotic elongation factor 1 alpha (EEF1A1 and EEF1A2) in pigs. Mendel University in Brno: Doctoral Thesis. (in Czech)
 
Svobodová Kateřina, Bílek Karel, Knoll Aleš (2008): Verification of reference genes for relative quantification of gene expression by real-time reverse transcription PCR in the pig. Journal of Applied Genetics, 49, 263-265  https://doi.org/10.1007/BF03195623
 
Vandesompele J., De Preter K., Pattyn F., Poppe B., Van Roy N., De Paepe A., Speleman F. (2002): Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology, 37, RESEARCH0034.
 
Wang Q.H., Chen J.Y., Guo K.N., Zheng X.B., Wu Q., Wei H., Liu Y. (2015): A correlation study of boar taint-related genes, boar taint substances and sex hormones in Bama miniature pigs at different ages. Indian Journal of Animal Research, 49, 745–751.
 
Xue J., Dial G.D., Pettigrew J.E. (1997): Performance, carcass and meat quality advantages of boars over barrows: A literature review. Journal of Swine Health and Production, 5, 21–28.
 
Zadinová Kateřina, Stupka Roman, Stratil Antonín, Čítek Jaroslav, Vehovský Karel, Lebedová Nicole, Šprysl Michal, Okrouhlá Monika (2017): Association analysis of SNPs in the porcine CYP2E1 gene with skatole, indole, and androstenone levels in backfat of a crossbred pig population. Meat Science, 131, 68-73  https://doi.org/10.1016/j.meatsci.2017.04.236
 
Zamaratskaia Galia, Rasmussen Martin Krøyer (2015): Immunocastration of Male Pigs – Situation Today. Procedia Food Science, 5, 324-327  https://doi.org/10.1016/j.profoo.2015.09.064
 
Zamaratskaia G., Gilmore W.J., Lundström K., Squires E.J. (2007): Effect of testicular steroids on catalytic activities of cytochrome P450 enzymes in porcine liver microsomes. Food and Chemical Toxicology, 45, 676-681  https://doi.org/10.1016/j.fct.2006.10.023
 
Zamaratskaia G., Zlabek V., Chen G., Madej A. (2009): Modulation of porcine cytochrome P450 enzyme activities by surgical castration and immunocastration. animal, 3, 1124-1132  https://doi.org/10.1017/S1751731109004510
 
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