Browse > Article
http://dx.doi.org/10.5713/ajas.2007.866

Use of the Non-electrophoretic Method to Detect Testis Specific Protein Gene for Sexing in Preimplantation Bovine Embryos  

Huang, Jinming (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences)
You, Wei (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences)
Wu, Naike (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences)
Tan, Xiuwen (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences)
Publication Information
Asian-Australasian Journal of Animal Sciences / v.20, no.6, 2007 , pp. 866-871 More about this Journal
Abstract
Testis-specific protein (TSPY) is a Y-specific gene, with up to 200 copy numbers in bulls. In order to make bovine embryo sexing under farm condition more feasible, the possibility of using a non-electrophoretic method to detect the TSPY gene for sexing bovine early embryos was examined. Primers were designed to amplify a portion of the TSPY gene and a common gene as an internal control primer. PCR optimization was carried out using a DNA template from bovine whole blood. Furthermore, embryo samples were diagnosed by this method and the sexing results were contrasted with those of the Loop-Mediated Isothermal Amplification (LAMP) method. The results showed that TSPY was as reliable a sexing method as LAMP. Forty-three morula and blastocyst embryos collected from superovulated donor dairy cattle were sexed by this method, and twenty-one embryos judged to be female embryos were transferred non-surgically to recipients 6 to 8 days after natural estrus. Out of 21 recipients, 9 were pregnant (42.86%) and all delivered female calves. The results showed that the sex predicted by this protocol was 100% accurate. In conclusion, the TSPY gene was a good male specific marker and indicated that a non-electrophoretic method was feasible and accurate to detect the TSPY gene for sexing preimplantation bovine embryos.
Keywords
Bovine Embryo; Sex Determination; TSPY Gene; PCR;
Citations & Related Records

Times Cited By Web Of Science : 2  (Related Records In Web of Science)
Times Cited By SCOPUS : 2
연도 인용수 순위
1 Chen,C. M., C. L. Hu, C. H. Wang, C. M. Hung, H. K. Wu, K. B. Choo and W. T. Cheng. 1999. Gender determination in single bovine blastomeres by polymerase chain reaction amplification of sex-specific polymorphic fragments in the amelogenin gene. Mol. Reprod. Dev. 54(3):209-214.   DOI   ScienceOn
2 Hasler, J. F., E. Cardey, J. E. Stokes and P. Bredbacka. 2002. Nonelectrophoretic PCR-sexing of bovine embryos in a commercial environment. Theriogenol. 58(8):1457-1469.   DOI   ScienceOn
3 Kageyama, S., I. Yoshida, K. Kawakura and K. Chikuni. 2004. A novel repeated sequence located on the bovine Y chromosome: its application to rapid and precise embryo sexing by PCR. J. Vet. Med. Sci. 66(5):509-514.   DOI   ScienceOn
4 Schempp, W., A. Binkele, J. Arnemann, B. Glaser, K. Ma, K. Taylor, R. Toder, J. Wolfe and A. C. Chandley. 1995. Comparative mapping of YRRM- and TSPY-related cosmids in man and hominoid apes. Chromosome Res. 3:227-234.   DOI   ScienceOn
5 Virta, J., J. Markola and J. Peippo. 2002. Sex determination of bovine embryo blastomeres by fluorogenic probes. Theriogenol. 57:2229-2236.   DOI   ScienceOn
6 Zeng, Y. T., M. L. Zhang and M. Y. Chen. 1992. Using the polymerase chain reaction (PCR) amplify the cow Y chromosome the sex determination area (SRY) sequence to sex cow embryo. Chinese Science Bulletin 37(5):97-98.
7 Bredbacka, P. 1998. Recent developments in embryo sexing and its field application. Reprod. Nutr. Dev. 38(6):605-613.   DOI   ScienceOn
8 Ennis, S. and T. F. Gallagher. 1994. A PCR-based sex-determination assay in cattle based on the bovine amelogenin locus. Anim. Genet. 25:425-427.   DOI   ScienceOn
9 Liu, Y. B. and J. M. Huang. 2004. Experiment of superovulation of Holstein heifer. J. China Herbivores. 24(01):14-16.
10 Chen, C. Y., L. S. Huang, J. B. Chen, N. S. Ding, L. H. Zhou and H. X. Xiao. 2003. Establishing and optimizing the system for sex determination of bovine preimplantation embryos. J. Agric. Biotech. 11(4):399-402.
11 Lindner, G. M. and R. W. Wright. 1983. Bovine embryo morphology and evaluation. Theriogenol. 20:407-416.   DOI   ScienceOn
12 Vogel, T., F. Dechend and E. Manz. 1997. Organization and expression of bovine TSPY. Mamm. Genome. 8:491-496.   DOI   ScienceOn
13 Chrenek, P. and J. Bulla. 2002. Simultaneous analysis of sex determination and K-casein genotypes from bovine preimplantation embryos. Czech J. Anim. Sci. 47(8):1-5.
14 Thibier, M. and M. Nibart. 1995. The sexing of bovine embryos in the field. Theriogenol. 43:71-80.   DOI   ScienceOn
15 Bredbacka, P., A. Kankaanpaa and J. Peippo. 1995. PCR-sexing of bovine embryos: A simplified protocol. Theriogenol. 44(2):167-176.   DOI   ScienceOn
16 Higuchi, R., G. Dollinger, P. S. Walsh and R. Griffith. 1992. Simultaneous amplification and detection of specific DNA sequences. Biotechnol. 10(4):413-417.   DOI   ScienceOn
17 Bradbury, M. W., L. M. Isola and J. W. Gordon. 1990. Enzymatic amplification of a Y chromosome repeat in a single blastomere allows identification of the sex of preimplantation mouse embryos. Proc. Natl. Acad. Sci. 87:4053-4057.   DOI   ScienceOn
18 Jakubiczka, S., F. Schnieders and J. Schmidtke. 1993. A bovine homologue of the human TSPY gene. Genomics. 17:732-735.   DOI   ScienceOn
19 Aasen, E. and J. F. Medrano. 1990. Amplification of the ZFY and ZFX genes for sex identification in humans, cattle, sheep and goats. Biotechnol. 8(12):1279-1281.   DOI   ScienceOn
20 Affara, N., C. Bishop and W. Brown. 1996. Report of the second international workshop on Y chromosome mapping. Cytogenet Cell Genet.73:33-76.   DOI
21 Shea, B. F. 1999. Determining the sex of bovine embryos using polymerase chain reaction results: A six-year study. Theriogenol. 51:841-854.   DOI   ScienceOn
22 Peura, T., J. M. Hyttinen, M. Turunen and J. Janne. 1991. A reliable sex determination assay for bovine preimplantation embryos using the polymerase chain reaction. Theriogenol. 35(3):547-555.   DOI   ScienceOn
23 Huang, J. M., J. Xiong, Y. B. Liu and W. You. 2005. Sex determination effect of bovine preimplantation embryo by LAMP method. Ecol. Dome. Anim. 26:20-22.
24 Machaty, Z., A. Paldi, T. Csaki, Z. Varga, I. Kiss, Z. Barandi and G. Vajta. 1992. Biopsy and sex determination by PCR of IVF bovine embryos. J. Reprod. Fertil. 98(2):467-470.
25 Pierce, K. E., J. E. Rice and J. A. Sanchez. 2000. Real-time PCR using molecular beacons for accurate detection of the Y chromosome in single human blastomere. Mol. Hum. Reprod. 6:1155-1164.   DOI   ScienceOn
26 Park, J. H., J. H. Lee, K. M. Choi, S. Y. Joung, J. Y. Kim, G. M. Chung, D. I. Jin and K. S. Im. 2001. Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. Theriogenol. 55(9):1843-1853.   DOI   ScienceOn
27 Utsumi, K., H. Takeda and M. Yamada. 1994. Sexing of bovine embryos by PCR using Sry primer. Theriogenol. 41:323(abstract).   DOI   ScienceOn
28 Manz, E., T. Vogel and P. Glatzel. 1998. Identification of an equine Y chromosome specific gene locus (eTSPY) with potential in preimplantation sex diagnosis.Theriogenol. 49:364.   DOI   ScienceOn
29 Hochman, D., Y. Zaron, I. Dekel, E. Feldmesser, J. F. Medrano, M. Shani and M. Ron. 1996. Multiple genotype analysis and sexing of IVF bovine embryos. Theriogenol. 46:1063-1075.   DOI   ScienceOn
30 Levinson, G., R. A. Fields, G. L. Harton, F. T. Parmer, A. Maddalena, E. F. Fugger and J. D. Schulman. 1992. Reliable gender screening for human preimplantation embryos using multiple DNA target sequences. Human Reprod. 7:1304-1313.   DOI
31 Plucienniczak, A. S. J. and J. Jaworski. 1982. Nucleotide sequence of bovine 1.715 satellite DNA and its relation to other bovine satellite sequences. J. Mol. Biol. 158:293-304.   DOI
32 Hirayama, H., S. Kageyama, S. Moriyasu, K. Sawai, S. Onoe, Y. Takahashi, S. Katagiri, K. Toen, K. Watanabe, T. Notomi, H. Yamashina, S .Matsuzaki and A. Minamihashi. 2004. Rapid sexing of bovine preimplantation embryos using loopmediated isothermal amplification. Theriogenol. 62(5):887-896.   DOI   ScienceOn
33 Gong, R. C., Y. O. Xu and R. Tang. 1997. ZFY gene sequences in bovine by PCR.J of Southwest University for Nationalities (Natural Science Edition). 17(3):234-237.