title: Dynamics of intracellular phospholipid membrane organization during oocyte maturation and successful vitrification of immature oocytes retrieved by ovum pick-up in cattle.
abstract: The objective was to determine if immature bovine oocytes with cumulus cells at the germinal vesicle (GV) stage could be vitrified by aluminum sheets (AS; pieces of sheet-like aluminum foil). Cleavage rates in fertilized oocytes previously vitrified by the AS procedure were higher than those vitrified by a nylon-mesh holder (NM) procedure (89.3 ± 2.1% vs. 65.0 ± 3.7%). Cleaved embryos derived from the AS but not from the NM procedures developed to blastocysts. Furthermore, to investigate the effects of vitrifying GV oocytes on cytoplasmic structure and on the ability to undergo cytoplasmic changes, the intracellular phospholipid membrane (IM) was stained with the lipophilic fluorescent dye, 3,3'-dioctadecyloxa-carbocyanine perchlorate. After vitrification by AS, the IM remained intact relative to that of oocytes vitrified by NM. During in vitro maturation, reorganization of the IM was also undamaged in oocytes vitrified by AS before oocyte maturation, and the IM within oocytes vitrified by the NM procedure was evidently impaired. Finally, vitrification (AS) was used for GV oocytes collected using the ovum pick-up method. A bull calf was born after in vitro production and subsequent embryo transfer. The vitrification techniques described herein should facilitate generation of viable in vitro production bovine blastocysts using oocytes recovered using the ovum pick-up method.
authors.affiliation: United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Aono
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nagatomo
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Takuma
authors.forename: Rika
authors.initials: R
authors.lastname: Nonaka
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Ono
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Wada
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Abe
authors.forename: Masashi
authors.initials: M
authors.lastname: Takahashi
authors.forename: Tomomasa
authors.initials: T
authors.lastname: Watanabe
authors.forename: Manabu
authors.initials: M
authors.lastname: Kawahara
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004624:Embryo Transfer / Q000662:veterinary; D005260:Female; D059471:In Vitro Oocyte Maturation Techniques; D007425:Intracellular Membranes / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D008297:Male; D008560:Membrane Fluidity / Q000502:physiology; D054315:Oocyte Retrieval / Q000379:methods*; D009865:Oocytes / Q000378:metabolism* / Q000502:physiology; D009866:Oogenesis / Q000502:physiology*; D010063:Ovum / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D010743:Phospholipids / Q000378:metabolism*; D011247:Pregnancy; D058989:Vitrification
chemical_list: D010743:Phospholipids