Sexing of mouse preimplantation embryos by detection of Y chromosome- specific sequences using polymerase chain reaction

Tetsuo Kunieda, M. Xian, E. Kobayashi, T. Imamichi, K. Moriwaki, Y. Toyoda

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

Detection of genes known to be present on the mammalian Y chromosome was adapted for sexing mouse early embryos using the polymerase chain reaction (PCR) method. Sry and Zfy genes located in the sex-determining region of the Y chromosome were chosen for Y-specific target sequences, and DXNds3 sequence on the X chromosome was chosen for control. The two-step PCR method using two pairs of primers for each of the target sequences was employed for detecting the sequences. When DNAs of male and female mice were amplified with these primers, male-specific fragments were detected even in DNAs that were equivalent in amount to two cells. Mouse embryos at the two-cell stage were separated into two individual blastomeres, and one blastomere was karyotyped at the second cleavage. The remaining blastomere was subjected to PCR amplification immediately or after having been cultured for 48 h up to the morula stage. The Sry and Zfy sequences were detected in about half the embryos; detection of the Sry and Zfy sequences corresponded exactly to the presence of the Y chromosome, except in one sample of male morula in which embryos may have been lost before the PCR amplification. It is concluded that the sex of mouse preimplantation embryos can be accurately determined through detection of the Y-specific sequences using the two-step PCR method, even with the single blastomeres separated at the two-cell stage.

Original languageEnglish
Pages (from-to)692-697
Number of pages6
JournalBiology of Reproduction
Volume46
Issue number4
Publication statusPublished - 1992
Externally publishedYes

Fingerprint

Y Chromosome
Blastocyst
Blastomeres
Polymerase Chain Reaction
Embryonic Structures
Morula
sry Genes
Mammalian Chromosomes
DNA
X Chromosome
Genes

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Sexing of mouse preimplantation embryos by detection of Y chromosome- specific sequences using polymerase chain reaction. / Kunieda, Tetsuo; Xian, M.; Kobayashi, E.; Imamichi, T.; Moriwaki, K.; Toyoda, Y.

In: Biology of Reproduction, Vol. 46, No. 4, 1992, p. 692-697.

Research output: Contribution to journalArticle

Kunieda, T, Xian, M, Kobayashi, E, Imamichi, T, Moriwaki, K & Toyoda, Y 1992, 'Sexing of mouse preimplantation embryos by detection of Y chromosome- specific sequences using polymerase chain reaction', Biology of Reproduction, vol. 46, no. 4, pp. 692-697.
Kunieda, Tetsuo ; 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. In: Biology of Reproduction. 1992 ; Vol. 46, No. 4. pp. 692-697.
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