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Frequency of sex chromosomal mosaicism in bovine embryos and its effects on sexing using a single blastomere by PCR

  • Jong Ho Lee (a1), Joong Hoon Park (a1), Eun Joo Choi (a2), Jong Taek Yoon (a2), Chang Sik Park (a3), Seong Ho Lee (a4), Kyung Soon Im (a1) and Dong Il Jin (a5)...

Abstract

Assessment of nuclear status is important when a biopsied single blastomere is used for embryo sexing. In this study we investigated the nuclear status of blastomeres derived from 8- to 16-cell stage in vitro fertilised bovine embryos to determine the representativeness of a single blastomere for embryo sexing. In 24 embryos analysed, the agreement in sex determination between a biopsied single blastomere and a matched blastocyst by polymerase chain reaction (PCR) was 83.3%. To clarify the discrepancies, karyotypes of blastomeres in 8- to 16-cell stage bovine embryos were analysed. We applied vinblastine sulfate at various concentrations and for different exposure times for metaphase plate induction in 8- to 16-cell stage bovine embryos. The 1.0 mg/ml vinblastine sulfate treatment for 15 h was selected as the most effective condition for induction of a metaphase plate (>45%). Among 22 embryos under these conditions, only 8 of 10 that had a normal diploid chromosome complement showed a sex chromosomal composition of XX or XY (36.4%) and 2 diploid embryos showed mosaicism of the opposite sex of XX and XY in blastomeres of the embryo (9.1%). One haploid embryo contained only one X-chromosome (4.5%). Four of another 11 embryos with a mixoploid chromosomal complement contained a haploid blastomere with a wrong sex chromosome (18.2%). In conclusion, assessment of nuclear status of 8- to 16-cell stage bovine embryos revealed that morphologically normal embryos had a considerable proportion of mixoploid blastomeres and sex chromosomal mosaicism; these could be the cause of discrepancies in the sex between biopsied single blastomeres and matched blastocysts by PCR.

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Corresponding author

All correspondence to: Dong Il Jin, Department of Applied Biological Science, Sun Moon University, Asan City, Chungnam, 338-708, Korea. Tel: +82 41 302285. Fax: +82 41 5417425. e-mail: dij1@sunmoon.ac.kr

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