title: Assessment of DNA damage in goat preantral follicles after vitrification of the ovarian cortex.
abstract: Effective methods for gamete preservation should have low impact on DNA integrity. The present study investigated the effects of vitrification of goat ovarian tissues on the occurrence of DNA fragmentation and DNA double-stand breaks using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) assay and detection of phosphorylated histone H2AX (γH2AX), respectively. Goat ovaries were collected at a local abattoir and 12 tissue fragments were prepared from each ovarian pair. Tissue fragments were used as fresh control samples or were cultured in vitro, vitrified or vitrified and cultured. Vitrification was performed using the Ovarian Tissue Cryosystem. Fragments from all groups (control and treatments) were processed for histology, transmission electron microscopy, TUNEL assay and immunofluorescence. Compared with fresh control samples, a lower percentage of morphologically normal follicles was detected in the vitrification followed by culture treatment group (P<0.05). Normal follicular ultrastructure was observed in all groups. Immunofluorescence revealed the presence of γH2AX foci in few oocytes and ovarian stromal cells. TUNEL-positive follicles were found in samples without significant differences among groups (P>0.05). In conclusion, the vitrification protocol used in the present study did not increase DNA damage in preantral follicles enclosed in goat ovarian tissues.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-930, Brazil.
authors.forename: Luciana R
authors.initials: LR
authors.lastname: Faustino
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-930, Brazil.
authors.forename: Adeline A
authors.initials: AA
authors.lastname: Carvalho
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-930, Brazil.
authors.forename: Cleidson M G
authors.initials: CM
authors.lastname: Silva
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-930, Brazil.
authors.forename: Rafael
authors.initials: R
authors.lastname: Rossetto
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-930, Brazil.
authors.forename: Cláudio A P
authors.initials: CA
authors.lastname: Lopes
authors.affiliation: Laboratory of Animal Physiology, Department of Animal Science, Federal University of Ceará, Av. Mister Hull, s/n Campus do Pici, Fortaleza, CE 60021-970, Brazil.
authors.forename: Maurício F
authors.initials: MF
authors.lastname: van Tilburg
authors.affiliation: Institute of Marine Science (LABOMAR), Federal University of Ceará, Av. Abolição, 3207, Meireles, Fortaleza, CE 60165-081, Brazil.
authors.forename: Pedro B M
authors.initials: PB
authors.lastname: Carneiro
authors.affiliation: Laboratory of Electron Microscopy, Department of Cell Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, Brasília, DF 70919-970, Brazil.
authors.forename: Sônia N
authors.initials: SN
authors.lastname: Báo
authors.affiliation: Laboratory of Animal Physiology, Department of Animal Science, Federal University of Ceará, Av. Mister Hull, s/n Campus do Pici, Fortaleza, CE 60021-970, Brazil.
authors.forename: Arlindo A A
authors.initials: AA
authors.lastname: Moura
authors.affiliation: Department of Animal Science, McGill University, 21,111 Lakeshore Road, Ste. Anne de Bellevue, Quebec, H9X 3V9, Canada.
authors.forename: Vilceu
authors.initials: V
authors.lastname: Bordignon
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-930, Brazil.
authors.forename: José R
authors.initials: JR
authors.lastname: Figueiredo
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-930, Brazil.
authors.forename: Ana Paula R
authors.initials: AP
authors.lastname: Rodrigues
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004249:DNA Damage / Q000187:drug effects*; D005260:Female; D006041:Goats; D006080:Ovarian Follicle / Q000187:drug effects; D010053:Ovary / Q000187:drug effects*; D014021:Tissue Preservation / Q000379:methods*; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents