Sperm Fertility of Transgenic Boar Harboring hEPO Gene is Decreased

  • Park Chun-Gyu (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Kim Sung-Woo (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Lee Poong-Yeon (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Han Joo-Hee (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Lee Hyun-Gi (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Byun Sung-June (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Yang Boh-Suk (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Lee Chang-Hyung (Department of Animal Science, Korea National Agricultural College, RDA) ;
  • Lee Hoon-Taek (Animal Resources Research Center, Konkuk University) ;
  • Chang Won-Kyong (Animal Biotechnology Division, National Livestock Research Institute) ;
  • Park Jin-Ki (Animal Biotechnology Division, National Livestock Research Institute)
  • Published : 2006.03.01

Abstract

This study was conducted to compare the reproduction ability of the wild type boar and recombinant human erythropoietin (hEPO) transgenic boar semen. Ejaculated boar semen was analyzed by flow cytometry, Elisa and IVF methods. In experiment 1, flow cytometric analysis showed that the live sperm ratio of transgenic boar sperm significantly lower (P<0.05) than that of wild type boar after incubation at 20, 22, 24 and 26 hr. In experiment 2, the presence and levels of various cytokines (IL-6, IL-10 and $TNF-{\alpha}$) to related animal reproduction in the seminal and blood plasma were examined using specific enzyme immunoassay. There was no significant difference between both groups. In experiment 3, the fertilizing capacity and developmental ability of both boar sperm were compared. The transgenic boar sperm had a significantly low capacity of penetration, sperm-zona binding, embryo development, and blastocyst formation compared to wild type sperm (P<0.05). These results suggest that transgenic boar sperm harboring hEPO gene has low sperm viability than wild type boar, and it is a reason to decrease of fertility and litter size.

Keywords

References

  1. Abeydeera LR, Wang WH, Cantley TC, Rieke A, Murphy CN, Prather RS, Day BN (2000): Development and viability of pig oocytes matured in a protein-free medium containing epidermal growth factor. Theriogenology 54:787-97 https://doi.org/10.1016/S0093-691X(00)00390-3
  2. Burdon T, Sankaran L, Wall RJ, Spencer M, Hennighausen L (1991): Expression of a whey acidic protein transgene during mammary development. J Biol Chem 11: 6909-6914
  3. Garner DL, Dobrinsky JR, Welch GR, Johnson LA (1996): Porcine sperm viability, oocyte fertilization and embryo development after staining spermatozoa with SYBR-14. Theriogenology 45:1103-1113 https://doi.org/10.1016/0093-691X(96)00067-2
  4. Evenson DP, Darzynkiewicz Z, Melamed MR (1980): Relation of mammalian sperm chromatin heterogeneity to fertility. Science 210:1131-1133 https://doi.org/10.1126/science.7444440
  5. Erslev A (1953): Humoral regulation of red cell production. Blood 8:349-357
  6. Feugang JM, Roover DE, Leonard S, Dessy F, Donnay I (2002): Kinetics of apoptosis in preimplantation bovine embryos produced in vitro and in vivo. Theriogenology 57:494 (Abstract)
  7. Bookfor FR, Schwarz LK (1991): Effects of interleukin- 6, interleukin-2, and tumor necrosis factor Q on transferring release from Sertoli cells in culture. Endocrinology 129:256-262 https://doi.org/10.1210/endo-129-1-256
  8. Garner DL, Johnson LA (1995): Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol Reprod 53:276-284 https://doi.org/10.1095/biolreprod53.2.276
  9. Gillan L, Evans G, Maxwell WM (2005): Flow cytometric evaluation of sperm parameters in relation to fertility potential. Theriogenology 63:445-457 https://doi.org/10.1016/j.theriogenology.2004.09.024
  10. Gjorret JO, Wengle J, King WA, Schellander K, Hyttel P (2002): Occurrence of apoptosis in bovine embryos reconstructed by nuclear transfer or derived in vivo. Theriogenology 57:495 (Abstract)
  11. Hogan B, Costantini F, Lacy E (1994): Manipulating the mouse embryo: a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory, pp127-216
  12. Eiserman J, Register KB, Striker RC, Collins JL (1989): The effect of tumor necrosis on human sperm motility in vitro. J Androl 10:270-274 https://doi.org/10.1002/j.1939-4640.1989.tb00100.x
  13. Jelkmann W (1992): Erythropoietin: structure, control of production, and function. Physiol Rev 72: 449-489 https://doi.org/10.1152/physrev.1992.72.2.449
  14. Johnson LA, Aalbers JG, Gooton HJG (1988): Artificial insemination of swine: fecundity of boar semen stored in Beltsville TS (BTS), modified Modena (MM) or MR-A and inseminated on one, three and four days after collection. Zuchthygiene (Berl) 23:49-55 https://doi.org/10.1111/j.1439-0531.1988.tb01080.x
  15. Lacombe C, Da Silva JL, Bruneval P, Camillery JP, Bariety J et al. (1988): Identification of tissues and cells producing erythropoietin in the anemic mouse. Contrib Nephrol 66:17-24
  16. Levy RR, Cordonier H, Czyba JC, Goerin JF (2001): Apoptosis in preimplantation mammalian embryo and genetics. Int J Anat Embryol 106(Suppl):2101-2108
  17. Limonta J, Pedraza A, Rodr Aguez A, Freyre M, Barral AM, Castro FO, Lleonart R, Garcia CA, Gavilondo JV, de la Fuente J (1995): Production of active anti-CD6 mouse/human chimeric antibodies in the milk of transgenic mice. Immunotechnology 1:107-113 https://doi.org/10.1016/1380-2933(95)00010-0
  18. Li S, Rosen JM (1994): Distal regulatory elements required for rat whey acidic protein gene expression in transgenic mice. J BioI Chem 269:14235-4243
  19. Abeydeera, LR, Day BN (1997): In vitro penetration of pig oocytes in a modified Tris-buffered medium: effect of BSA, caffeine and calcium. Theriogenology 48:537-544 https://doi.org/10.1016/S0093-691X(97)00270-7
  20. Harrison RAP (1997): Sperm plasma membrane characteristics and boar semen fertility. J Reprod Fertil (Suppl) 52:271-283 https://doi.org/10.1530/jrf.0.0520271
  21. Mattioli M, Bacci ML, Galeati G, Seren E (1989): Developmental competence of pig oocytes matured and fertilized in vitro. Theriogenology 31:1201-1207 https://doi.org/10.1016/0093-691X(89)90089-7
  22. Miller LJ, Fischer KA, Goralnick SJ, Litt M, Burleson JA, Albertsen P, Kreutzer DL (2002): Interleukin- 10 levels in seminal plasma: implications for chronic prostatitis-chronic pelvic pain syndrome. J Urol 167:753-756 https://doi.org/10.1016/S0022-5347(01)69139-0
  23. Skinner MK (1991): Cell-cell interactions in the testis. Endocrine Rev 12:45-77 https://doi.org/10.1210/edrv-12-1-45
  24. Rajasekaran M, Hellstrom WJG, Naz RK, Sikka SC (1995): Oxidative stress and interleukins in seminal plasma dUring leukocytospermia. Fertil Steril 64: 166-171
  25. Gruschwitz MS, Bresinschek R, Bresinschek HP (1996): Cytokine levels in the seminal plasma of infertile males. J Androl 14:158-163
  26. Park JK, Lee YK, Lee P, Chung HJ, Kim S, Lee HG, Seo MK, Han JH, Park CG, Kim HT, Kim YK, Min KS, Kim JH, Lee HT, Chang WK (2006): Recombinant human erythropoietin produced in milk of transgenic pigs. J Biotechnol (in press)
  27. Pierce CN, Larson DF (2005) Inflammatory cytokine inhibition of erythropoiesis in patients implanted with a mechanical circulatory assist device. Perfusion 20(2):83-90 https://doi.org/10.1191/0267659105pf793oa
  28. Paleyanda RK, Zhang DW, Hennighausen L, McKnight RA, Lubon H (1994): Regulation of human protein C gene expression by the mouse WAP promoter. Transgenic Res 3:335-343 https://doi.org/10.1007/BF01976765
  29. Kelly RW (1995): Immunosupressive mechanisms in semen : Implications for contraception. Hum Reprod 10:1686-1693 https://doi.org/10.1093/oxfordjournals.humrep.a136156
  30. Tanphaichitr N, Smith J, Mongkolsirikieart S, Gradil C, Lingwood C (1993): Role of a gamete specific sulfoglycolipid-immobilizing protein on mouse sperm- egg binding. Dev BioI 156:164-175 https://doi.org/10.1006/dbio.1993.1067
  31. Vera O, Vasquez LA, Munoz MG (2003): Semen quality and presence of cytokines in seminal fluid of bull ejaculates. Theriogenology 60:553-558 https://doi.org/10.1016/S0093-691X(03)00031-1
  32. Wall RJ, Rexroad CE, Powell A, Shamay A, McKnight R, Hennighausen L (1996): Synthesis and secretion of the mouse whey acidic protein in transgenic sheep. Transgenic Res 5:67-72 https://doi.org/10.1007/BF01979923
  33. Yamauchi-Takihara, Ihara Y, Ogata A, Yoshizaki K, Azuma J, Kishimoto T (1995): Hypoxic stress induces cardiac myocyte-derived interleukin-6. Circulation 91:1520-1524 https://doi.org/10.1161/01.CIR.91.5.1520
  34. Yoshida M, Ishizaki Y, Kawagishi H (1990): Blastocyst formation by pig embryos resulting from invitro fertilization of oocytes matured in vitro. J Reprod Fertil 88:1-8 https://doi.org/10.1530/jrf.0.0880001
  35. Zhang J, Gao J (2004): Determination of IL-1beta, IL-4 and IL-10 contents in the seminal plasma of infertile patients and its clinical value. Zhonghua Nan Ke Xue 10:851-4
  36. Zhu Y, Osterlundh I, Hulten F, Magnusson U (2004): Tumor necrosis factor-alpha, interleukin-6, serum amyloid A, haptoglobin, and cortisol concentrations in sows following intrarrlarrunary inoculation of Escherichia coli. Am J Vet Res 65:1434-1439 https://doi.org/10.2460/ajvr.2004.65.1434