Molecular Cytogenetics of Human Single Pronucleated Zygotes

The aim of the present study was to use fluorescence in situ hybridization to analyze the chromosome status of zygotes with a single pronucleus from in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment cycles. In addition, we performed immunocytochemical detection of nuclear lamins and histone H3 trimethylated at lysine-9, Me(3)H3K9. Zygotes were processed 24 hours after insemination or injection to assure the absence of asynchrony. In opposition to previous results, we observed 2 pronuclei in 16 of 18 IVF zygotes and 40 of 64 ICSI zygotes, suggesting premature pronuclear breakdown. In IVF and ICSI zygotes, the rate of normal diploidy was only 6 of 16 and 27 of 56, respectively, suggesting that monopronucleated zygotes should not be used in assisted reproductive treatments. The possible mechanisms are discussed and compared to previous studies of monopronucleated zygotes.

[1]  A. Veiga,et al.  In vitro development and chromosome constitution of embryos derived from monopronucleated zygotes after intracytoplasmic sperm injection. , 2013, Fertility and sterility.

[2]  M. F. Cardoso,et al.  Impact of GnRH ovarian stimulation protocols on intracytoplasmic sperm injection outcomes , 2009, Reproductive biology and endocrinology : RB&E.

[3]  D. Chardonnens,et al.  Removal of spermatozoa with externalized phosphatidylserine from sperm preparation in human assisted medical procreation: effects on viability, motility and mitochondrial membrane potential , 2009, Reproductive biology and endocrinology : RB&E.

[4]  P. de Boer,et al.  Parental origin of chromatin in human monopronuclear zygotes revealed by asymmetric histone methylation patterns, differs between IVF and ICSI , 2009, Molecular reproduction and development.

[5]  M. Herbert,et al.  Can ‘abnormally’ fertilized zygotes give rise to viable embryos? , 2006, Human fertility.

[6]  W. Reik,et al.  Dynamic chromatin modifications characterise the first cell cycle in mouse embryos. , 2005, Developmental biology.

[7]  B. Imthurn,et al.  Chromosome analysis of single- and multipronucleated human zygotes proceeded after the intracytoplasmic sperm injection procedure , 1996, Journal of Assisted Reproduction and Genetics.

[8]  Jacques Cohen,et al.  Origin of single pronucleated human zygotes , 1993, Journal of Assisted Reproduction and Genetics.

[9]  B. Behr,et al.  Monozygotic Twin Birth After the Transfer of a Cleavage Stage Embryo Resulting from a Single Pronucleated Oocyte , 2004, Journal of Assisted Reproduction and Genetics.

[10]  M. Sousa,et al.  Preimplantation genetic diagnosis using FISH for carriers of Robertsonian translocations: the Portuguese experience , 2002, Prenatal diagnosis.

[11]  R. Goldman,et al.  Nuclear lamins: building blocks of nuclear architecture. , 2002, Genes & development.

[12]  D. Gilbert,et al.  Heterochromatin, HP1 and methylation at lysine 9 of histone H3 in animals , 2002, Chromosoma.

[13]  D. Gardner,et al.  Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. , 2000, Fertility and sterility.

[14]  P. de Boer,et al.  Distribution of Atr protein in primary spermatocytes of a mouse chromosomal mutant: a comparison of preparation techniques , 2000, Chromosoma.

[15]  A. Trounson,et al.  Pregnancy and birth resulting from transfer of a blastocyst observed to have one pronucleus at the time of examination for fertilization. , 1999, Human reproduction.

[16]  J Cohen,et al.  Chromosome abnormalities in human embryos. , 1998, Human reproduction update.

[17]  P. Devroey,et al.  Successful preimplantation genetic diagnosis is related to the number of available cumulus-oocyte complexes. , 1998, Human reproduction.

[18]  D. Payne,et al.  Preliminary observations on polar body extrusion and pronuclear formation in human oocytes using time-lapse video cinematography. , 1997, Human reproduction.

[19]  A. V. Van Steirteghem,et al.  The chromosomal constitution of embryos developing from abnormally fertilized oocytes after intracytoplasmic sperm injection and conventional in-vitro fertilization. , 1997, Human reproduction.

[20]  P. Braude,et al.  The timing of pronuclear formation, DNA synthesis and cleavage in the human 1-cell embryo. , 1996, Molecular human reproduction.

[21]  D. Payne,et al.  Aetiology of failed and abnormal fertilization after intracytoplasmic sperm injection. , 1995, Human reproduction.

[22]  M. Sousa,et al.  Key elements of a highly efficient intracytoplasmic sperm injection technique: Ca2+ fluxes and oocyte cytoplasmic dislocation. , 1995, Fertility and sterility.

[23]  Z. Rosenwaks,et al.  Male and female genomes associated in a single pronucleus in human zygotes. , 1995, Biology of reproduction.

[24]  K. Sultan,et al.  Chromosomal status of uni-pronuclear human zygotes following in-vitro fertilization and intracytoplasmic sperm injection. , 1995, Human reproduction.

[25]  Mário Sousa,et al.  Fertiliza1tion and early embryology: Ultrastructural analysis of fertilization failure after intracytoplasmic sperm injection , 1994 .

[26]  Z. Nagy,et al.  Time-course of oocyte activation, pronucleus formation and cleavage in human oocytes fertilized by intracytoplasmic sperm injection. , 1994, Human reproduction.

[27]  J. Harper,et al.  Identification of the sex of human preimplantation embryos in two hours using an improved spreading method and fluorescent in-situ hybridization (FISH) using directly labelled probes. , 1994, Human reproduction.

[28]  E. Coonen,et al.  Optimal preparation of preimplantation embryo interphase nuclei for analysis by fluorescence in-situ hybridization. , 1994, Human reproduction.

[29]  M. Sousa,et al.  Ultrastructural analysis of fertilization failure after intracytoplasmic sperm injection. , 1994, Human reproduction.

[30]  J. Squire,et al.  Characterization of abnormal one pronuclear human oocytes by morphology, cytogenetics and in-situ hybridization. , 1993, Human reproduction.

[31]  P. Devroey,et al.  Cytogenetic and morphological observations of single pronucleated human oocytes after in-vitro fertilization. , 1993, Human reproduction.

[32]  J. Mandelbaum,et al.  Cytogenetic analysis and developmental capacity of normal and abnormal embryos after IVF. , 1989, Human reproduction.

[33]  A. Tarkowski.  An Air-Drying Method for Chromosome Preparations from Mouse Eggs , 1966 .