Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Gene Expression Analysis of Pregnant Specific Stage in the Miniature Pig Ovary

  • Yun, Seong-Jo (Animal Biotechnology, Graduate School of Bio. & Information Technology, Institute of Genetic Engineering, Hankyong National University) ;
  • Noh, Won-Gun (Animal Biotechnology, Graduate School of Bio. & Information Technology, Institute of Genetic Engineering, Hankyong National University) ;
  • Yoon, Jong-Taek (Animal Biotechnology, Graduate School of Bio. & Information Technology, Institute of Genetic Engineering, Hankyong National University) ;
  • Min, Kwan-Sik (Animal Biotechnology, Graduate School of Bio. & Information Technology, Institute of Genetic Engineering, Hankyong National University)
  • Published : 2009.12.31
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Abstract

The miniature pig is considered to be a better organ donor breed for xenotransplantation than other pig breeds because the size of the organs of the miniature pig is similar to that of humans. In this study, we aimed at identifying differentially expressed genes in the miniature pig ovary during pregnancy. For this, we used the miniature pig ovary model, annealing control primer-based reverse transcription polymerase chain reaction (PCR), quantitative real-time PCR (qRT-PCR), and northern blotting analysis. We identified 13 genes showing differential expression on the based of pregnancy status and validated 8 genes using qRT-PCR. We also sequenced the full-length cDNA of ephrin receptor A4 (EphA4), which had a significant difference in expression level, and validated it by northern blotting. These genes may provide a better understanding of the cellular and molecular mechanisms during pregnancy in miniature pig ovary.

Keywords

References

  1. Adams RH, Wilkinson GA, Weiss C, Diella F, Gale NW, Deutsch U, Risau W, Klein R (1999): Roles of ephrinB ligands and EphB receptors in cardiovascular development: demarcation of arterial/venous domains, vascular morphogenesis, and sprouting angiogenesis. Genes Dev 13:295-306 https://doi.org/10.1101/gad.13.3.295
  2. Auersperg N, Wong AS, Choi KC, Kang SK, Leung PC (2001): Ovarian surface epithelium: biology, endocrinology, and pathology. Endocrine Reviews 22: 255-288 https://doi.org/10.1210/er.22.2.255
  3. Bazer FW, Ott TL, Spencer TE (1998): Endocrinology of the transition from recurring estrous cycles to establishment of pregnancy in subprimate mammals. In: Bazer FW, editor. The Endocrinology of Pregnancy. Totowa: Humana Press, pp 1-34
  4. Dor FJ, Tseng YL, Cheng J, Moran K, Sanderson TM, Lancos CJ, Shimizu A, Yamade K, Awwad M, Sachs DH, Hawley RJ, Schuurman HJM, Cooper DK (2004): Alpha 1,3-galactosyltransferase gene-knockout miniature swine produce natural cytotoxic antiGal antibodies. Transplantation 78:15-20
  5. Frisen J, Holmberg J, Barbacid M (1999): Ephrins and their Eph receptors: multitalented directors of embryonic development. EMBO J 18:5159-5165 https://doi.org/10.1093/emboj/18.19.5159
  6. Gerety SS, Wang HU, Chen ZF, Anderson DJ (1999): Symmetrical mutant phenotypes of the receptor EphB4 and its specific transmembrane ligand ephrin-B2 in cardiovascular development. Mol Cell 4: 403-414
  7. Goldshmit Y, Galea MP, Bartlett PF, Turnley AM (2006): EphA4 regulates central nervous system vascular formation. The Journal of Comparative Neurology 497:864-875 https://doi.org/10.1002/cne.21029
  8. Golovan SP, Hakimov HA, Verschoor CP, Walters S, Gadish M, Elsik C, Schenkel F, Chiu DKY, Forsberg CW (2008): Analysis of Sus scrofa liver proteome and identification of proteins differentially expressed between genders, and conventional and genetically enhanced lines. Comparative Biochemis and Physiology Part D 3:234-242 https://doi.org/10.1016/j.cbd.2008.05.001
  9. Hwang IT, Kim YJ, Kim SH, Kwak CI, Gu YY, Chun JY (2003): Annealing control primer system for improving specificity of PCR amplification. Biotechniques 35:1180-1184
  10. Ka HH, Seo HW, Kim MG, Choi YH, Lee CK (2009): Identification of differentially expressed genes in the uterine endometrium on day 12 of the estrous cycle and pregnancy in pigs. Molecular Reproduction & Development 76:75-84 https://doi.org/10.1002/mrd.20935
  11. Kim YJ, Kwak CI, Gu YY, Hwang IT, Chun JY (2004): Annealing control primer system for identification of differentially expressed genes on agarose gels. Biotechniques 36:424-426
  12. Lai L, Kolber-Simonds D, Park KW, Cheong HT, Greenstein JL, Im GS, Samuel M, Bonk A, Rieke A, Day BN, Murphy CN, Carter DB, Hawley RJ, Prather RS (2002): Production of alpha-1,3-galactosyltransferase knockout pigs by nuclear transfer cloning. Science 295:1089-1092 https://doi.org/10.1126/science.1068228
  13. Madej A, Lang A, Brandt Y, Kindahl H, Madsen MT, Einarsson S (2005): Factors regulating ovarian function in pigs. Domestic Animal Endocrinology 29:347-361 https://doi.org/10.1016/j.domaniend.2005.02.030
  14. Ok JK, Park HJ, Kwon DK, Kang JT, Jang G, Lee BC (2009): Effect of recipient breed on delivery rate of cloned miniature pig. Zygote 17:203-207 https://doi.org/10.1017/S0967199409005267
  15. Rajeevan MS, Ranamukhaarachchi DG, Vernon DG, Unger ER (2001): Use of real-time quantitative PCR to validate the results of cDNA array and differential display PCR technologies. Methods 25:443–451 https://doi.org/10.1006/meth.2001.1266
  16. Ramaekers F, Smedts F, Vooijs GP (1992): Keratins as differentiation markers in tumor biology and surgical pathology, in Spandidos D (ed): Current Perspectives in Molecular and Cellular Oncology. JAI Press, London, England, pp 285-318
  17. Reynolds LP, Killilea SD, Redmer DA (1992): Angiogenesis in the female reproductive system. FASEB Journal 6:886-892 https://doi.org/10.1096/fasebj.6.3.1371260
  18. Ricke WA, Redmer DA, Reynolds LP (1995): Initial characterization of mitogenic factors produced by porcine corpora lutea throughout the estrous cycle. Biology of Reproduction 52 (Supplement 1):112
  19. Papp LV, Lu J, Holmgren A, Khanna KK (2007): From selenium to selenoproteins: synthesis, identity, and their role in human health. Antioxid Redox Signal 9:775-806 https://doi.org/10.1089/ars.2007.1528.
  20. Tsunoda Y, Kato Y (2002): Recent progress and problems in animal cloning. Differentiation 69:158-161 https://doi.org/10.1046/j.1432-0436.2002.690405.x
  21. Wang HU, Chen ZF, Anderson DJ (1998): Molecular distinction and angiogenic interaction between embryonic arteries and veins revealed by ephrin- B2 and its receptor Eph-B4. Cell 93:741-753 https://doi.org/10.1016/S0092-8674(00)81436-1
  22. Wang Y, Spitz MR, Tsou AM-H, Zhang K, Makan N, Wu X (2002): Sulfotransferase (SULT) 1A1 polymorphism as a predisposition factor for lung cancer: a case-control analysis. Lung Cancer 35:137-142 https://doi.org/10.1016/S0169-5002(01)00406-8
  23. Wauters CCAP, Smedts F, Gerrits LGM, Bosman F, Ramaekers FCS (1995): Keratins 7 and 20 as diagnostic markers of carcinomas metastatic to the ovary. Human Pathology 26:852-855 https://doi.org/10.1016/0046-8177(95)90006-3