Sex determination in single mouse blastomeres by polymerase chain reaction
Abstract
In sex determination of mammalian preimplantation embryos, viability of biopsied embryos and accuracy of sexing are together important. In consideration of this point, single blastomeres were mechanically isolated from mouse embryos using a micromanipulator and then sexed by polymerase chain reaction (PCR) using mouse Y chromosome-specific primers. All of 260 embryos biopsied at the four-cell and morula stage survived. Developmental rate of the embryos to normal blastocysts was 93 and 94%, respectively. Sex determination of single blastomeres was performed by amplification of a mouse Y chromosome-specific DNA sequence using PCR technique. The ratio of male to female embryos was 53 and 47%, respectively. The sex-determined embryos were transferred to the uteri of pseudopregnant recipients to test the consistency of the assay system. The sex of 27 of 29 mice developed from male and female embryos agreed with the predicted sex. The method developed for embryo biopsy and sexing could be used for diagnosis of defective genes at the stage of the preimplantation embryos of human and other domestic animals.
Key words
embryo biopsy single blastomeres polymerase chain reaction sexingPreview
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References
- 1.King WA: Sexing embryos by cytological methods. Theriogenology 1984;21:7–17Google Scholar
- 2.Monk M, Handyside AH: Sexing of preimplantation mouse embryos by measurement of X-linked gene dosage in a single blastomere. J Reprod Fert 1988;82:365–368Google Scholar
- 3.Wachtel SS: H-Y antigen in the study of sex determination and control of sex ratio. Theriogenology 1984;21:18–28Google Scholar
- 4.Avery B: Impact of asynchronous ovulations on the expression of sex-dependent growth rate in bovine preimplantation embryos. J Reprod Fert 1989;87:627–631Google Scholar
- 5.Xu KP, Yadav BR, King WA, Betteridge KJ: Sex-related differences in developmental rates of bovine embryos produced and cultured In Vitro. Mol Reprod Dev 1992;31:249–252PubMedGoogle Scholar
- 6.Bondioli KR, Ellis SB, Pryor JH, Williams MW, Harpold MM: The use of male-specific chromosomal DNA fragments to determine the sex of bovine preimplantation embryos. Theriogenology 1989;31:95–104Google Scholar
- 7.Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA: Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988;239:487–491PubMedGoogle Scholar
- 8.Bradbury MW, Isola LM, Gordon JW: Enzymatic amplification of a Y chromosome repeat in a single blastomere allows identification of the sex of preimplantation mouse embryo. Proc Natl Acad Sci USA 1990;87:4053–4057PubMedGoogle Scholar
- 9.Kunieda T, Xian M, Kobayashi E, Imamichi T, Moriwaki K, Toyoda Y: Sexing of mouse preimplantation embryos by detection of Y chromosome-specific sequences using polymerase chain reaction. Biol Reprod 1992;46:692–697PubMedGoogle Scholar
- 10.Handyside AH, Kontogianni EH, Hardy K, Winston RML: Pregnancies from biopsied human preimplantation embryos sexed by Y-specific DNA amplification. Nature 1990;344:768–770PubMedGoogle Scholar
- 11.Herr CM, Matthaei KI, Petrzak U, Reed KC: A rapid Y-chromosome-detecting ovine embryo sexing assay. Theriogenology 1990;33:245Google Scholar
- 12.Herr CM, Holt NA, Matthaei KI, Reed KC: Sex of progeny from bovine embryos sexed with a rapid Y-chromosomedetection assay. Theriogenology 1990;33:247Google Scholar
- 13.Peura T, Hyttinen J-M, Turunen M, Janne J: A reliable sex determination assay for bovine preimplantation embryos using the polymerase chain reaction. Theriogenology 1991;35:547–555Google Scholar
- 14.Quinn P, Barros C, Whittingham DG: Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa. J Reprod Fert 1982;66:161–168PubMedGoogle Scholar
- 15.Gordon JW, Gang I: Use of zona drilling for safe and effective biopsy of murine oocytes and embryos. Biol Reprod 1990;42:869–876PubMedGoogle Scholar
- 16.Whittingham DG: Culture of mouse ova. J Reprod Fert (Suppl) 1971;14:7–21Google Scholar
- 17.Gubbay J, Collignon J, Koopman P, Capel B, Economou A, Munsterberg A, Vivian N, Goodfellow P, Lovell-Badge R: A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Nature 1990;346:245–250PubMedGoogle Scholar
- 18.Garrisi GJ, Talansky BE, Grunfeld L, Sapira V, Navot D, Gordon JW: Clinical evaluation of three approaches to micromanipulation-assisted fertilization. Fertil Steril 1990;54:671–677PubMedGoogle Scholar
- 19.Malter HE, Cohen J: Partial zona dissection of the human oocyte: A nontraumatic method using micromanipulation to assist zona pellucida penetration. Fertil Steril 1989;51:139–148PubMedGoogle Scholar
- 20.Picard L, King WA, Betteridge KJ: Production of sexed calves from frozen-thawed embryos. Vet Rec 1985;117:603–608PubMedGoogle Scholar
- 21.Strom CM, Rechitsky S, Verlinsky Y: Reliability of gender determination using the polymerase chain reaction (PCR) for single cells. J Vitro Fert Embryo Transfer 1991;8:225–229Google Scholar
- 22.Grifo JA, Tang YX, Cohen J, Gilbert F, Sanyal MK, Rosenwaks Z: Pregnancy after embryo biopsy and coamplification of DNA from X and Y chromosomes. JAMA 1992;268:727–729PubMedGoogle Scholar