title: The use of antifreeze protein type III for vitrification of in vitro matured bovine oocytes.
abstract: The aim of this study was to evaluate the use of antifreeze protein type III (AFP III) into vitrification medium on meiotic spindle morphology of in vitro matured bovine oocytes as well as the fertilization and blastocyst rates. Mature cumulus-oocyte complexes (COC) were distributed in four groups: control (untreated), vitrified without supplementation (AFP0) or supplemented with 500 (AFP500) or 1000 ng/mL (AFP1000) into vitrification solutions. Samples from each group were used to analyze the organization of meiotic spindle by confocal microscopy and the remaining COC were submitted to in vitro fertilization and culture for eight days. Control group exhibited only 15% of abnormal spindle. However, the spindle morphology was affected in all vitrified groups regardless to AFP concentration: 75.8%, 76.1% and 69.2% (P > 0.05) for AFP0, AFP500and AFP1000, respectively. Similar cleavage rate was obtained among the vitrified groups (AFP0= 17.9%, AFP500= 16.9% and AFP1000= 17.8%), but lower (P < 0.05) compared with control group (68.7%). At Day 5 of culture, embryo production rate of AFP500(30.8%) and AFP1000(25.0%) were similar to control group (49.4%). However, at Day 8 of culture, AFP0,AFP500and AFP1000groups exhibited lower (P < 0.05) blastocyst rates (10.0%, 3.8% and 9.4%, respectively) when compared to control (41.1%). In conclusion, AFP III did not preserve meiotic spindle organization against the cryoinjuries. However, the use of AFP III improved embryo development at Day 5 of culture, although this effect was not maintained up to the blastocyst formation.
authors.affiliation: Laboratory of Physiology and Control of Reproduction, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: Dowglish F
authors.initials: DF
authors.lastname: Chaves
authors.affiliation: Laboratory of Physiology and Control of Reproduction, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: Iana S
authors.initials: IS
authors.lastname: Campelo
authors.affiliation: Laboratory of Physiology and Control of Reproduction, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: Mirelly M A S
authors.initials: MMAS
authors.lastname: Silva
authors.affiliation: Laboratory of Physiology and Control of Reproduction, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: Maajid H
authors.initials: MH
authors.lastname: Bhat
authors.affiliation: Laboratory of Physiology and Control of Reproduction, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: Darcio I A
authors.initials: DIA
authors.lastname: Teixeira
authors.affiliation: Laboratory of Physiology and Control of Reproduction, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: Luciana M
authors.initials: LM
authors.lastname: Melo
authors.affiliation: Faculty of Veterinary Medicine, Fluminense Federal University, Niterói, RJ, Brazil.
authors.forename: Joanna M G
authors.initials: JMG
authors.lastname: Souza-Fabjan
authors.affiliation: INRA, Physiologie de la Reproduction et des Comportements, Nouzilly, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Mermillod
authors.affiliation: Laboratory of Physiology and Control of Reproduction, State University of Ceará, Fortaleza, CE, Brazil. Electronic address: vicente.freitas@uece.br.
authors.forename: Vicente J F
authors.initials: VJF
authors.lastname: Freitas
mesh_terms: D000818:Animals; D021301:Antifreeze Proteins / Q000494:pharmacology*; D001755:Blastocyst / Q000187:drug effects; D002417:Cattle; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro / Q000187:drug effects / Q000379:methods; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods*; D018613:Microscopy, Confocal; D009865:Oocytes / Q000378:metabolism; D058989:Vitrification
chemical_list: D021301:Antifreeze Proteins; D003451:Cryoprotective Agents
keywords: AFP III; Cryopreservation; Embryo production; Meiotic spindle; Oocyte