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Analysis of ENPP2 in the Uterine Endometrium of Pigs Carrying Somatic Cell Nuclear Transfer Cloned Embryos

  • Seo, Heewon ;
  • Choi, Yohan ;
  • Yu, Inkyu ;
  • Shim, Jangsoo ;
  • Lee, Chang-Kyu ;
  • Hyun, Sang-Hwan ;
  • Lee, Eunsong ;
  • Ka, Hakhyun
  • Received : 2013.03.18
  • Accepted : 2013.06.04
  • Published : 2013.09.01

Abstract

Somatic cell nuclear transfer (SCNT) is a useful tool for animal cloning, but the efficiency of producing viable offspring by SCNT is very low. To improve this efficiency in the production of cloned pigs, it is critical to understand the interactions between uterine function and cloned embryos during implantation. Lysophosphatidic acid (LPA) is a lipid mediator that plays an important role in the establishment of pregnancy in pigs; however, LPA production in the uterine endometrium of pigs carrying SCNT-cloned conceptuses has not been determined. Therefore, we investigated expression of ENPP2, an LPA-generating enzyme, in the uterine endometrium of gilts with conceptuses derived from SCNT during the implantation period. Uterine endometrial tissue and uterine flushing were obtained from gilts carrying SCNT-derived conceptuses and from gilts carrying conceptuses resulting from natural mating on d 12 of pregnancy. Our results demonstrated no difference in the level of ENPP2 mRNA expression in the uterine endometrium between gilts carrying SCNT-derived conceptuses and gilts carrying naturally-conceived conceptuses, but secretion of ENPP2 protein into the uterine lumen did decrease significantly in pigs with SCNT-derived conceptuses. These results indicate that expression and secretion of ENPP2, which are critical for appropriate LPA production and successful pregnancy, are dysregulated in the uterine endometrium of pigs carrying SCNT-derived conceptuses.

Keywords

Pig;Pregnancy;ENPP2;LPA;SCNT

References

  1. Bauersachs, S., S. E. Ulbrich, V. Zakhartchenko, M. Minten, M. Reichenbach, H. D. Reichenbach, H. Blum, T. E. Spencer, and E. Wolf. 2009. The endometrium responds differently to cloned versus fertilized embryos. Proc. Natl. Acad. Sci. USA 106:5681-5686. https://doi.org/10.1073/pnas.0811841106
  2. Burghardt, R. C., J. A. Bowen, G. R. Newton, and F. W. Bazer. 1997. Extracellular matrix and the implantation cascade in pigs. J. Reprod. Fertil. Suppl 52:151-164.
  3. Campbell, K. H., R. Alberio, I. Choi, P. Fisher, R. D. Kelly, J. H. Lee, and W. Maalouf. 2005. Cloning: eight years after Dolly. Reprod. Domest. Anim. 40:256-268. https://doi.org/10.1111/j.1439-0531.2005.00591.x
  4. Chae, J. I., S. K. Cho, J. W. Seo, T. S. Yoon, K. S. Lee, J. H. Kim, K. K. Lee, Y. M. Han, and K. Yu. 2006. Proteomic analysis of the extraembryonic tissue from cloned porcine embryos. Mol. Cell. Proteomics 5:1559-1566. https://doi.org/10.1074/mcp.M500427-MCP200
  5. Gardell, S. E., A. E. Dubin, and J. Chun. 2006. Emerging medicinal roles for lysophospholipid signaling. Trends Mol. Med. 12:65-75. https://doi.org/10.1016/j.molmed.2005.12.001
  6. Geisert, R. D. and J. V. Yelich. 1997. Regulation of conceptus development and attachment in pigs. J. Reprod. Fertil. Suppl. 52:133-149.
  7. Gerrard, J. M. and P. Robinson. 1989. Identification of the molecular species of lysophosphatidic acid produced when platelets are stimulated by thrombin. Biochim. Biophys. Acta 1001:282-285. https://doi.org/10.1016/0005-2760(89)90112-4
  8. Ishii, I., N. Fukushima, X. Ye, and J. Chun. 2004. Lysophospholipid receptors: signaling and biology. Annu. Rev. Biochem. 73:321-354. https://doi.org/10.1146/annurev.biochem.73.011303.073731
  9. Jaeger, L. A., G. A. Johnson, H. Ka, J. G. Garlow, R. C. Burghardt, T. E. Spencer, and F. W. Bazer. 2001. Functional analysis of autocrine and paracrine signalling at the uterine-conceptus interface in pigs. Reprod. Suppl. 58:191-207.
  10. Jouneau, A., Q. Zhou, A. Camus, V. Brochard, L. Maulny, J. Collignon, and J. P. Renard. 2006. Developmental abnormalities of NT mouse embryos appear early after implantation. Development 133:1597-1607. https://doi.org/10.1242/dev.02317
  11. Ka, H., H. Seo, M. Kim, S. Moon, H. Kim, and C. K. Lee. 2008. Gene expression profiling of the uterus with embryos cloned by somatic cell nuclear transfer on day 30 of pregnancy. Anim. Reprod. Sci. 108:79-91. https://doi.org/10.1016/j.anireprosci.2007.07.008
  12. Aoki, J., A. Inoue, and S. Okudaira. 2008. Two pathways for lysophosphatidic acid production. Biochim. Biophys. Acta 1781:513-518. https://doi.org/10.1016/j.bbalip.2008.06.005
  13. Bazer, F. W., T. L. Ott, and T. E. Spencer. 1998. Endocrinology of the transition from recurring estrous cycles to establishment of pregnancy in subprimate mammals. In: Endocrinology of pregnancy (Ed. F. W. Bazer). Totowa, NJ: Humana Press 1-34.
  14. Ka, H., T. E. Spencer, G. A. Johnson, and F. W. Bazer. 2000. Keratinocyte growth factor: expression by endometrial epithelia of the porcine uterus. Biol. Reprod. 62:1772-1778. https://doi.org/10.1095/biolreprod62.6.1772
  15. Kim, H. R., J. K. Kang, J. T. Yoon, H. H. Seong, J. K. Jung, H. M. Lee, C. S. Park, and D. I. Jin. 2005. Protein profiles of bovine placenta derived from somatic cell nuclear transfer. Proteomics 5:4264-4273. https://doi.org/10.1002/pmic.200401297
  16. Kim, M., H. Seo, Y. Choi, W. Hwang, C. K. Lee, and H. Ka. 2009. Aberrant expression of retinol-binding protein, osteopontin and fibroblast growth factor 7 in the porcine uterine endometrium of pregnant recipients carrying embryos produced by somatic cell nuclear transfer. Anim. Reprod. Sci. 112:172-181. https://doi.org/10.1016/j.anireprosci.2008.04.029
  17. Lee, R. S., A. J. Peterson, M. J. Donnison, S. Ravelich, A. M. Ledgard, N. Li, J. E. Oliver, A. L. Miller, F. C. Tucker, B. Breier, and D. N. Wells. 2004. Cloned cattle fetuses with the same nuclear genetics are more variable than contemporary half-siblings resulting from artificial insemination and exhibit fetal and placental growth deregulation even in the first trimester. Biol. Reprod. 70:1-11.
  18. Liszewska, E., P. Reinaud, E. Billon-Denis, O. Dubois, P. Robin, and G. Charpigny. 2009. Lysophosphatidic acid signaling during embryo development in sheep: involvement in prostaglandin synthesis. Endocrinology 150:422-434. https://doi.org/10.1210/en.2008-0749
  19. Livak, K. J. and T. D. Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402-408. https://doi.org/10.1006/meth.2001.1262
  20. Mansouri-Attia, N., O. Sandra, J. Aubert, S. Degrelle, R. E. Everts, C. Giraud-Delville, Y. Heyman, L. Galio, I. Hue, X. Yang, X. C. Tian, H. A. Lewin, and J. P. Renard. 2009. Endometrium as an early sensor of in vitro embryo manipulation technologies. Proc. Natl. Acad. Sci. USA 106:5687-5692. https://doi.org/10.1073/pnas.0812722106
  21. Pope, W. F. 1988. Uterine asynchrony: a cause of embryonic loss. Biol. Reprod. 39:999-1003. https://doi.org/10.1095/biolreprod39.5.999
  22. Pope, W. F., M. S. Lawyer, B. S. Nara, and N. L. First. 1986. Effect of asynchronous superinduction on embryo survival and range of blastocyst development in swine. Biol. Reprod. 35:133-137. https://doi.org/10.1095/biolreprod35.1.133
  23. Seo, H. and H. Ka. 2011. Expression of lysophosphatidic acid receptor 3 in the uterine endometrium of pigs with somatic cell nuclear transfer cloned conceptuses. J. Anim. Sci. Technol. 53:203-209. https://doi.org/10.5187/JAST.2011.53.3.203
  24. Seo, H., M. Kim, Y. Choi, C. K. Lee, and H. Ka. 2008. Analysis of lysophosphatidic acid (LPA) receptor and LPA-induced endometrial prostaglandin-endoperoxide synthase 2 expression in the porcine uterus. Endocrinology 149:6166-6175. https://doi.org/10.1210/en.2008-0354
  25. Seo, H., Y. Choi, J. Shim, M. Kim, and H. Ka. 2012. Analysis of the lysophosphatidic acid-generating enzyme ENPP2 in the uterus during pregnancy in pigs. Biol. Reprod. 87:77. https://doi.org/10.1095/biolreprod.112.099564
  26. Seo, H., Y. Choi, J. Shim, Y. Choi, and H. Ka. 2012. Regulatory mechanism for expression of IL1B receptors in the uterine endometrium and effects of IL1B on prostaglandin synthetic enzymes during the implantation period in pigs. Biol. Reprod. 87:31. https://doi.org/10.1095/biolreprod.112.099051
  27. Spencer, T. E. and F. W. Bazer. 2004. Uterine and placental factors regulating conceptus growth in domestic animals. J. Anim. Sci. 82 E-Suppl:E4-13.
  28. Tigyi, G. and R. Miledi. 1992. Lysophosphatidates bound to serum albumin activate membrane currents in Xenopus oocytes and neurite retraction in PC12 pheochromocytoma cells. J. Biol. Chem. 267:21360-21367.
  29. Tokumura, A., E. Majima, Y. Kariya, K. Tominaga, K. Kogure, K. Yasuda, and K. Fukuzawa. 2002. Identification of human plasma lysophospholipase D, a lysophosphatidic acid-producing enzyme, as autotaxin, a multifunctional phosphodiesterase. J. Biol. Chem. 277:39436-39442. https://doi.org/10.1074/jbc.M205623200
  30. Tokumura, A., Y. Kanaya, M. Miyake, S. Yamano, M. Irahara, and K. Fukuzawa. 2002. Increased production of bioactive lysophosphatidic acid by serum lysophospholipase D in human pregnancy. Biol. Reprod. 67:1386-1392. https://doi.org/10.1095/biolreprod.102.004051
  31. Tokumura, A., T. Kume, S. Taira, K. Yasuda, and H. Kanzaki. 2009. Altered activity of lysophospholipase D, which produces bioactive lysophosphatidic acid and choline, in serum from women with pathological pregnancy. Mol. Hum. Reprod. 15:301-310. https://doi.org/10.1093/molehr/gap017
  32. Tranguch, S., T. Daikoku, Y. Guo, H. Wang, and S. K. Dey. 2005. Molecular complexity in establishing uterine receptivity and implantation. Cell. Mol. Life Sci. 62:1964-1973. https://doi.org/10.1007/s00018-005-5230-0
  33. Wells, D. N. 2005. Animal cloning: problems and prospects. Rev. Sci. Tech. 24:251-264.
  34. Ye, X., K. Hama, J. J. Contos, B. Anliker, A. Inoue, M. K. Skinner, H. Suzuki, T. Amano, G. Kennedy, H. Arai, J. Aoki, and J. Chun. 2005. LPA3-mediated lysophosphatidic acid signalling in embryo implantation and spacing. Nature 435:104-108. https://doi.org/10.1038/nature03505

Acknowledgement

Supported by : Rural Development Administration, National Research Foundation