Vet Med - Czech, 2007, 52(12):540-546 | DOI: 10.17221/1884-VETMED

Lower quality bovine embryos may be successfully used for sex determination

M. Lopatarova1, S. Cech1, P. Krontorad2, L. Holy1, J. Hlavicova1, R. Dolezel1
1 University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic
2 BOVET Company, Sloupnice, Czech Republic

The aim of this study was to evaluate the effect of sex determination procedures in Day 7 to Day 8 bovine embryos of various quality. For the purposes of comparison we used high quality (HQ) as well as lower quality (LQ) embryos obtained from superovulated donors. The healthy embryonic cells isolated from HQ embryos and blastomeres protruding into the perivitelline space of LQ embryos, were analysed by polymerase chain reactions (PCR) using primers specific for the Y chromosome determinant. After microsurgical intervention and completion of sex determination, the female embryos were then transferred (ET) to synchronized recipients. A total of 310 embryos of HQ were used and gender was safely determined in 275 cases (88.7%). PCR analysis of extruded cells isolated from 170 LQ embryos was carried out with certainty only in 111 embryos (65.3%, P < 0.01). After ET of 122 HQ sex defined embryos, pregnancy was established in 69 recipients (56.6%). A similar conception rate 51.9% (27/52) was found after the ET of sex defined embryos designated as LQ. The accuracy of analysis was confirmed after calving and revealed that designated female sex coincided with 95.5% and 96.2% of calves when HQ and LQ embryos were transferred, respectively. Our results clearly show that a microsurgical technique in combination with PCR method represents a rapid and reliable approach for sex determination in HQ as well as in LQ preimplantation bovine embryos and can be used in field conditions for the regulation of the sex of progeny in selected herds.

Keywords: cattle; embryo; superovulation; sex determination; polymerase chain reaction; embryo transfer

Published: December 31, 2007  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Lopatarova M, Cech S, Krontorad P, Holy L, Hlavicova J, Dolezel R. Lower quality bovine embryos may be successfully used for sex determination. Vet Med - Czech. 2007;52(12):540-546. doi: 10.17221/1884-VETMED.
Download citation

References

  1. Bondioli K.R., Ellis S.B., Pryor J.H., Williams M.W., Harpold M.M. (1989): The use of male-specific chromosomal DNA fragments to determine the sex of bovine preimplantation embryos. Theriogenology, 31, 95-104. Go to original source...
  2. Booman P., Kruijt L., Veerhuis R., Hengst A.M., Tiemann M., Ruch F.E. (1989): Sexing bovine embryos with monoclonal antibodies against the H-Y antigen. Livestock Production Science, 23, 1-16. Go to original source...
  3. Bredbacka P., Vemala R., Peippo J., Bredbacka K. (1994): Survival of biopsied and sexed bovine demi-embryos. Theriogenology, 41, 1023-1031. Go to original source... Go to PubMed...
  4. Chrenek P., Boulanger L., Heyman Y., Uhrin P., Laurincik J., Bulla J., Renard J.P. (2001): Sexing and multiple genotype analysis from a single cell of bovine embryo. Theriogenology, 55, 1071-1081. Go to original source... Go to PubMed...
  5. Edwards R.G., Gardner R.L. (1967): Sexing of live rabbit blastocyst. Nature, 214, 576-577. Go to original source... Go to PubMed...
  6. Ekici H., Turan N., Sontas B.H., Helps C.R., Senunver A., Ylmaz H. (2006): Sex determination of bovine embryos using polymerase chain reaction (PCR). Revue de Medicine Veterinaire, 157, 441-444.
  7. Herr C.M., Holt N.A., Matthaei K.I., Reed K.C. (1990): Sex of progeny from bovine embryos sexed with a rapid Y-chromosome-detection assay. Theriogenology, 33, 247. Go to original source...
  8. Herr C.M., Steel T., Lascelles A., Reed K.C. (1995): Instruction Manual: Splitting, Biopsy & Rapid Sexing of Cattle Embryos. AB Technology, Inc., Pullmann, WA. 58 pp.
  9. Hirayama H., Kageyama S., Moriyasu S., Sawai K., Onoe S., Takahashi Y., Katagiri S., Toen K., Watanabe K., Notomi T., Yamashina H., Matzuzaki S., Minamihashi A. (2004): Rapid sexing of bovine preimplantation embryo using loop-mediated isothermal amplification. Theriogenology, 62, 887-896. Go to original source... Go to PubMed...
  10. Holy L., Lopatarova M., Krontorad P., Holy J. (1990): Systematic use of embryo transfer in cattle under condition of high animal concentration (in German). Monatsheft Veterinaermedizin, 45, 409-413.
  11. Kobayashi J., Sekimoto A., Uchida H., Wada T., Sasaki K., Sasada H., Umezu M., Sato E. (1998): Rapid detection of male-specific DNA sequence in bovine embryos using fluorescence in situ hybridization. Molecular Reproduction and Development, 51, 390-394. Go to original source... Go to PubMed...
  12. Lee J.H., Park J.H., Lee S.H., Park C.S., Jin D.I. (2004): Sexing using single blastomere derived from IVF bovine embryos by fluorescence in situ hybridization (FISH). Theriogenology, 62, 1452-1458. Go to original source... Go to PubMed...
  13. Leonhard M., Korszenbaum M., Cotinot C., Chesne P., Heyman Y., Stinnakre M.G., Bishop C., Delouis C., Vaiman M., Fellous M. (1987): Sexing bovine embryos using Y chromosome specific DNA probe. Theriogenology, 27, 248. Go to original source...
  14. Lopatarova M., Holy L., Vinkler A. (2001): Effect on survival of micromanipulating the zona pellucida of bovine embryos. Acta Veterinaria Brunensis, 70, 49-56. Go to original source...
  15. Lopatarova M., Krontorad P., Holy L., Zajic J. (2006): Determining the sex of early bovine embryo in practical conditions (in Czech). Veterinarstvi, 56, 696-699.
  16. Lopes R.F.F., Forell F., Oliveira A.T.D., Rodrigues J.L. (2001): Splitting and biopsy for bovine embryo sexing under field conditions. Theriogenology, 56, 1383-1392. Go to original source... Go to PubMed...
  17. Machaty Z., Paldi A., Csaki T., Varga Z., Kiss I., Barandi Z., Vajta G. (1993): Biopsy and sex determination by PCR of IVF bovine embryos. Journal of Reproduction and Fertility, 98, 467-470. Go to original source... Go to PubMed...
  18. Park J.H., Lee J.H., Choi K.M., Joung S.Y., Kim J.Y., Chung G.M., Jin D.I., Im K.S. (2001): Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. Theriogenology, 55, 843-853. Go to original source... Go to PubMed...
  19. Rall W.F., Leibo S.P. (1987): Production of sexed bovine pregnancies by cytogenetic analysis of cultured demiembryos. Theriogenology, 27, 269. Go to original source...
  20. Ramalho M.F.P.D.T., Garcia J.M., Esper C.R., Vandini R., Alves B.C.A., Almeina I.L. Jr., Hossepian de Lima V.F.M., Moreira-Filho C.A. (2004): Sexing of murine and bovine embryo by developmental arrest induced by high-titer H-Y antisera. Theriogenology, 62, 1569-1576. Go to original source... Go to PubMed...
  21. Schroder A., Roschlau D., Giehm D., Schwerin M., Thomsen P.D. (1990): Sex determination of bovine embryos using Y-specific primers in the polymerase chain reaction. Archiv fur Tierzucht, 33, 293-299.
  22. Shea B.F. (1999): Determining the sex of bovine embryos using polymerase chain reaction results: a six-year retrospective study. Theriogenology, 51, 841-854. Go to original source... Go to PubMed...
  23. Singh E.L., Hare W.C.D. (1980): The feasibility of sexing bovine morula stage embryos prior to embryo transfer. Theriogenology, 14, 421-427. Go to original source...
  24. Taneja M., Rao K.B.C.A., Gangawane S., Zawar S.G., Totey S.M. (1998): Rapid sexing of bovine preimplantation embryos using polymerase chain reaction: production of calves with predetermined sex under field conditions. Indian Journal of Experimental Biology, 36, 1201-1208.
  25. Thibier M., Nibart M. (1995): The sexing of bovine embryos in the field. Theriogenology, 43, 71-80. Go to original source...
  26. Tominaga K., Hamada Y. (2004): Efficient production of sex-identified and cryosurvived bovine in-vitro produced blastocysts. Theriogenology, 61, 1181-1191. Go to original source... Go to PubMed...
  27. White K.L., Lindner G.M., Anderson G.B., BonDurant R.H. (1982): Survival after transfer of "sexed" mouse embryos exposed to H-Y antisera. Theriogenology, 18, 655-662. Go to original source...
  28. Williams T.J. (1986): A technique for sexing mouse embryos by a visual colorimetric assay of the X-linked enzyme, glucose 6-phosphate dehydrogenase. Theriogenology, 25, 733-739. Go to original source... Go to PubMed...
  29. Yu W., Li S., Fang J., Sun X., Cui L., Fu J., Bai Y., Fang Y., Shangguan B. (2006): Field studies on the effectiveness of the YCD embryo sexing technique in bovine. Reproduction, Fertility and Development, 19, 299-300. Go to original source...

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.