title: Short-term preservation of canine preantral follicles: Effects of temperature, medium and time.
abstract: The use of the large pool of preantral follicles is a promising alternative to provide high numbers of fertilizable oocytes to reproductive biotechnology. This issue is particularly important to canids, since current rates of success of in vitro techniques using oocytes are very limited, and many species within this family are threatened by extinction. The aim of this study was to evaluate effects of temperature, medium and time on morphology and viability of canine preantral follicles during short-term preservation. Canine ovaries were cut into fragments which were incubated in 0.9% NaCl solution or in minimum essential medium (MEM) at 4, 20 or 38 degrees C for 2, 6, 12 or 24 h. Afterwards, preantral follicles were analyzed by histology, transmission electron microscopy and viability testing using trypan blue, calcein-AM and ethidium homodimer-1. Percentages of morphological normal and viable follicles were maintained similar to control (time 0 h) after incubation in 0.9% NaCl at 4 or 20 degrees C for up to 6h and at 38 degrees C for 2 h. Using MEM, such preservation was possible for 12h at 4 or 20 degrees C, and for 6h at 38 degrees C. These results indicate that preservation of canine preantral follicles might be better accomplished through hypothermic (4 or 20 degrees C) storage in MEM, which ensures maintenance of morphology and viability for up to 12h.
authors.affiliation: Faculty of Veterinary, PPGCV, LAMOFOPA, State University of Ceará, Av. Paranjana 1700, Campus do Itaperi, CEP 60.740-903, Fortaleza, CE, Brazil. lopes@izw-berlin.de
authors.forename: Cláudio Afonso Pinho
authors.initials: CA
authors.lastname: Lopes
authors.forename: Regiane Rodrigues
authors.initials: RR
authors.lastname: dos Santos
authors.forename: Juliana Jales de Hollanda
authors.initials: JJ
authors.lastname: Celestino
authors.forename: Mônica Aline Parente
authors.initials: MA
authors.lastname: Melo
authors.forename: Roberta Nogueira
authors.initials: RN
authors.lastname: Chaves
authors.forename: Claudio Cabral
authors.initials: CC
authors.lastname: Campello
authors.forename: José Roberto Viana
authors.initials: JR
authors.lastname: Silva
authors.forename: Sônia Nair
authors.initials: SN
authors.lastname: Báo
authors.forename: Katarina
authors.initials: K
authors.lastname: Jewgenow
authors.forename: José Ricardo
authors.initials: JR
authors.lastname: de Figueiredo
mesh_terms: D000818:Animals; D002470:Cell Survival; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D006107:Granulosa Cells / Q000648:ultrastructure; D008928:Mitochondria / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000648:ultrastructure; D009865:Oocytes / Q000502:physiology*; D009926:Organ Preservation / Q000379:methods / Q000662:veterinary; D006080:Ovarian Follicle / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D010052:Ovariectomy / Q000662:veterinary