title: Next Generation Sequencing-Based Comprehensive Chromosome Screening in Mouse Polar Bodies, Oocytes, and Embryos.
abstract: Advanced reproductive age is unequivocally associated with increased aneuploidy in human oocytes, which contributes to infertility, miscarriages, and birth defects. The frequency of meiotic chromosome segregation errors in oocytes derived from reproductively aged mice appears to be similar to that observed in humans, but a limitation of this important model system is our inability to accurately identify chromosome-specific aneuploidy. Here we report the validation and application of a new low-pass whole-genome sequencing approach to comprehensively screen chromosome aneuploidy in individual mouse oocytes and blastocysts. First, we validated this approach by using single mouse embryonic fibroblasts engineered to have stable trisomy 16. We further validated this method by identifying reciprocal chromosome segregation errors in the products of meiosis I (gamete and polar body) in oocytes from reproductively aged mice. Finally, we applied this technology to investigate the incidence of aneuploidy in blastocysts derived from in vitro- and in vivo-matured oocytes in both young and reproductively aged mice. Using this next generation sequencing approach, we quantitatively assessed meiotic and mitotic segregation errors at the single chromosome level, distinguished between errors due to premature separation of sister chromatids and classical nondisjunction of homologous chromosomes, and quantified mitochondrial DNA (mtDNA) segregation in individual cells. This whole-genome sequencing technique, therefore, greatly improves the utility of the mouse model system for the study of aneuploidy and is a powerful quantitative tool with which to examine the molecular underpinnings of mammalian gamete and early embryo chromosome segregation in the context of reproductive aging and beyond.
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, Reproductive Medicine Associates of New Jersey, Basking Ridge, New Jersey Rutgers-Robert Wood Johnson Medical School, New Brunswick, New Jersey ntreff@rmanj.com.
authors.forename: Nathan R
authors.initials: NR
authors.lastname: Treff
authors.affiliation: National Foundation for Fertility Research, Lone Tree, Colorado.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Krisher
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, Reproductive Medicine Associates of New Jersey, Basking Ridge, New Jersey.
authors.forename: Xin
authors.initials: X
authors.lastname: Tao
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, Reproductive Medicine Associates of New Jersey, Basking Ridge, New Jersey.
authors.forename: Heather
authors.initials: H
authors.lastname: Garnsey
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, Reproductive Medicine Associates of New Jersey, Basking Ridge, New Jersey.
authors.forename: Chelsea
authors.initials: C
authors.lastname: Bohrer
authors.affiliation: National Foundation for Fertility Research, Lone Tree, Colorado.
authors.forename: Elena
authors.initials: E
authors.lastname: Silva
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, Reproductive Medicine Associates of New Jersey, Basking Ridge, New Jersey Rutgers-Robert Wood Johnson Medical School, New Brunswick, New Jersey.
authors.forename: Jessica
authors.initials: J
authors.lastname: Landis
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, Reproductive Medicine Associates of New Jersey, Basking Ridge, New Jersey Rutgers-Robert Wood Johnson Medical School, New Brunswick, New Jersey.
authors.forename: Deanne
authors.initials: D
authors.lastname: Taylor
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, Reproductive Medicine Associates of New Jersey, Basking Ridge, New Jersey Rutgers-Robert Wood Johnson Medical School, New Brunswick, New Jersey.
authors.forename: Richard T
authors.initials: RT
authors.lastname: Scott
authors.affiliation: Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
authors.forename: Teresa K
authors.initials: TK
authors.lastname: Woodruff
authors.affiliation: Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas fduncan@kumc.edu.
authors.forename: Francesca E
authors.initials: FE
authors.lastname: Duncan
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000737:chemistry; D002460:Cell Line; D002885:Chromosomes, Human, Pair 16; D056915:DNA Copy Number Variations; D004272:DNA, Mitochondrial / Q000032:analysis*; D004622:Embryo, Mammalian / Q000737:chemistry; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D008297:Male; D051379:Mice; D009030:Mosaicism; D009630:Nondisjunction, Genetic; D009865:Oocytes / Q000737:chemistry; D059705:Polar Bodies / Q000737:chemistry; D017422:Sequence Analysis, DNA / Q000379:methods*; D014314:Trisomy / Q000175:diagnosis*
chemical_list: D004272:DNA, Mitochondrial
keywords: aging; aneuploidy; embryo; oocyte; whole-genome sequencing