Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Study on the Reproductive Function in Transgenic Pig Harboring Human Erythropoietin (hEPO) Gene

  • Lee, Hyun-Gi (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Hwi-Cheul (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Chung, Hak-Jae (Hanwoo Experimental Station, National Institute of Animal Science, RDA) ;
  • Hwang, In-Sul (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Choi, Myoung-Seob (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Byun, Sung-June (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Seung-Hoon (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Kim, Min-Ji (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Woo, Jae-Seok (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Chang, Won-Kyong (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Poong-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Lee, Hoon-Taek (Department of Animal Science, KonKuk University) ;
  • Park, Jin-Ki (Animal Biotechnology Division, National Institute of Animal Science, RDA)
  • Published : 2008.06.30
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Abstract

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.

Keywords

References

  1. Anagnostou A, Liu Z, Steiner M, Chin K, Lee ES, Kessimian N, Noguchi CT (1994): Erythropoietin receptor mRNA expression in human endothelial cells. Proc Natl Acad Sci USA 91:3974-3978
  2. Ashley RA, Dubuque SH, Dvorak B, Woodward SS, Williams SK, Kling PJ (2002): Erythropoietin stimulates vasculogenesis in neonatal rat mesenteric microvascular endothelial cells. Pediatr Res 51:472-478 https://doi.org/10.1203/00006450-200204000-00012
  3. Carlini RG, Reyes AA, Rothstein M (1995): Recombinant human erythropoietin stimulates angiogenesis in vitro. Kidney Int 47:740-745 https://doi.org/10.1038/ki.1995.113
  4. Fisher JW (1997): Erythropoietin: physiologic and pharmacologic aspects. Proc Soc Exp Biol Med 216: 358-369
  5. Hanahan D, Folkman J (1996): Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86:353-364 https://doi.org/10.1016/S0092-8674(00)80108-7
  6. Heeschen C, Aicher A, Lehmann R, Fichtlscherer S, Vasa M, Urbich C, Mildner-Rihm C, Martin H, Zeiher AM, Dimmeler S (2003): Erythropoietin is a potent physiologic stimulus for endothelial progenitor cell mobilization. Blood 102:1340-1346 https://doi.org/10.1182/blood-2003-01-0223
  7. Horiguchi H, Oguma E, Kayama F (2005): The effects of iron deficiency on estradiol-induced suppression of erythropoietin induction in rats: Implications of pregnancy-related anemia. Blood 106:67-74 https://doi.org/10.1182/blood-2004-06-2350
  8. Horsley BR, Estienne MJ, Harper AF, Purcell SH, Baitis HK, Beal WE, Knight JW (2005): Effect of PG 600 on the timing of ovulation in gilts treated with altrenogest. J Anim Sci 83:1690-1695 https://doi.org/10.2527/2005.8371690x
  9. Kim MO, Kim SH, Shin MJ, Lee DB, Kim TW, Kim KS, Ha JH, Lee S, Park YB, Kim SJ, Ryoo ZY (2007): Human erythropoietin induces lung failure and erythrocytosis in transgenic mice. Mol Cells 23: 17-22
  10. Macmillan D, Bill RM, Sage KA, Fern D, Flitsch SL (2001): Selective in vitro glycosylation of recombinant proteins: semi-synthesis of novel homogeneous glycoforms of human erythropoietin. Chem Biol 8: 133-145 https://doi.org/10.1016/S1074-5521(00)90065-6
  11. Muller-Ehmsen J, Schmidt A, Krausgrill B, Schwinger RH, Bloch W (2006): Role of erythropoietin for angiogenesis and vasculogenesis: From embryonic development through adulthood. Am J Physiol Heart Circ Physiol 290:H331-340 https://doi.org/10.1152/ajpheart.01269.2004
  12. Paleyanda RK, Zhang DW, Hennighausen L, Mc- Knight 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
  13. Park CG, Kim S, Lee P, Han JH, Lee HG, Byun SJ, Yang BS, Lee CH, Lee HT, Chang WK, Park JK (2006a): Sperm Fertility of Transgenic Boar Harboring hEPO Gene is Decreased. Reprod Dev Biol 30:27-34
  14. 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 (2006b): Recombinant human erythropoietin produced in milk of transgenic pigs. J Biotechnol 122:362-371 https://doi.org/10.1016/j.jbiotec.2005.11.021
  15. Pittius CW, Hennighausen L, Lee E, Westphal H, Nicols E, Vitale J, Gordon K (1988): A milk protein gene promoter directs the expression of human tissue plasminogen activator cDNA to the mammary gland in transgenic mice. Proc Natl Acad Sci USA 85:5874-5878
  16. Ribatti D, Presta M, Vacca A, Ria R, Giuliani R, Dell'Era P, Nico B, Roncali L, Dammacco F (1999): Human erythropoietin induces a pro-angiogenic phenotype in cultured endothelial cells and stimulates neovascularization in vivo. Blood 93:2627-2636
  17. Sasaki H, Bothner B, Dell A, Fukuda M (1987): Carbohydrate structure of erythropoietin expressed in Chinese hamster ovary cells by a human erythropoietin cDNA. J Biol Chem 262:12059-12076
  18. Sasaki R (2003): Pleiotropic functions of erythropoietin. Intern Med 42:142-149 https://doi.org/10.2169/internalmedicine.42.142
  19. Signoret JP (1971): The mating behaviour of the sow. In: Cole, D.J.A.(eds), Pig production. Butterworths, UK, pp 295-313
  20. Yasuda Y, Masuda S, Chikuma M, Inoue K, Nagao M, Sasaki R (1998): Estrogen-dependent production of erythropoietin in uterus and its implication in uterine angiogenesis. J Biol Chem 273:25381-25387 https://doi.org/10.1074/jbc.273.39.25381