Taiwanese Journal of Obstetrics and Gynecology
Volume 45, Issue 4 , Pages 313-316, December 2006

Distinction Between Paternal and Maternal Contributions to the Tripronucleus in Human Zygotes Obtained after In Vitro Fertilization

  • San-Li Yu

      Affiliations

    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan
    • ,
  • Robert Kuo-Kuang Lee

      Affiliations

    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan
    • Division of Reproduction and Endocrinology, Department of Medical Research, Mackay Memorial Hospital, Tamshui, Taiwan
    • Corresponding Author InformationCorrespondence to: Dr Robert Kuo-Kuang Lee, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, 92, Section 2, Chung Shan North Road, Taipei 104, Taiwan
    • ,
  • Jin-Tsung Su

      Affiliations

    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan
    • ,
  • Yu-Fen Chih

      Affiliations

    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan
    • ,
  • Yi-Chun Tsai

      Affiliations

    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan
    • ,
  • Ming-Huei Lin

      Affiliations

    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan
    • ,
  • Yuh-Ming Hwu

      Affiliations

    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Mackay Memorial Hospital, Taipei, Taiwan

Accepted 9 June 2006.

Article Outline

Summary 

Objective

The aim of this study was to determine the genetic composition of tripronuclear (3PN) zygotes. Fluorescence in situ hybridization (FISH) was used to quantify the sperm tails in human 3PN zygotes.

Materials and Methods

Ovarian hyperstimulation was performed using a standard long protocol consisting of gonadotropin-releasing hormone agonist (GnRHa) with human menopausal gonadotropins (HMG) and/or recombinant follicle stimulating hormone (rFSH). Human chorionic gonadotropin (hCG) was administered when at least two leading follicles reached 18 mm in mean diameter. Oocytes were retrieved from the follicles transvaginally under ultrasound guidance 34-36 hours after administration of hCG. The oocytes were examined 16-18 hours after fertilization for the presence and number of pronuclei. The 3PN zygotes were then fixed and extracted in buffer at 37°C for 60 minutes. After washing in a blocking solution, the 3PN zygotes were prepared for indirect immunofluorescence using monoclonal antibody and Hoechst dye 33342 to visualize the number of pronuclei.

Results

Twenty-one 3PN zygotes were randomly collected for evaluation. The number of 3PN zygotes containing one, two or four sperm tails were two (10%), 18 (85%) and one (5%), respectively.

Conclusion

Based on our observations, additional pronuclei are mainly of paternal origin.

Key Words:  dispermy , FISH , fluorescence in situ hybridization , tripronuclear zygotes

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PII: S1028-4559(09)60249-7

doi:10.1016/S1028-4559(09)60249-7

Taiwanese Journal of Obstetrics and Gynecology
Volume 45, Issue 4 , Pages 313-316, December 2006

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