The study on X-Y spirmpreseparation and Testie determining gene

1993 Fiscal Year Final Research Report Summary

• Project/Area Number: 04671021
• Research Category: Grant-in-Aid for General Scientific Research (C)
• Allocation Type: Single-year Grants
• Research Field: Obstetrics and gynecology
• Research Institution: Keio University
• Principal Investigator: KOBAYASHI Toshifumi Keio University School of Medicine Associate professor, 医学部・産婦人科, 助教授 (30051460)
• Co-Investigator(Kenkyū-buntansha): HARA Izumi Keio University School of Medicine Assistant, 医学部・産婦人科, 助手 (90208645) ; SUEOKA Kou Keio University School of Medicine Assistant, 医学部・産婦人科, 助教授 (90162833)
• Project Period (FY): 1992 – 1993
• Keywords: X-Y sperm ppresepatation / SRY gene / PCR / FISH / DXZ-1 / DYZ-1 / percoll / density gradient centrifugation

Research Abstract

The patients of sex-linked genetic deisorders have been disired to be curedby the teatment of preselection at the stage of gametes and the selection at the early embryos. Specially the objective of this study is the development of sperm preselection procedures and establishment of the detemination method for sperm X and Y using DNA probe analysisi prior to its selection. The distribution of X and Y sperm has been demonstrated by fluorescence in situ hybridization (FISH) nethod. The decondensation precedure of the sperm nuclei was established. The sperm unclei have to be treated with diethiothreitol (DTT) for getting DNA probe through nuclear membrane. DXY-1 probe and DYZ-1 have been used as specific DNA probe for X and Y sperm respectively. The development of sperm sex analysis has been directed to PCR amplification method for the purpose of reducing necessary numbers of sperm cells. The sex determinig lesion Y(SRY) gene was used to determine the existence of sperm. although final objective should be the analysisi from single sperm cell, it is still of the reducing process of sperm cell umbers.