title: Modulation of aquaporins 3 and 9 after exposure of ovine ovarian tissue to cryoprotectants followed by in vitro culture.
abstract: Our aim has been to evaluate the effect of cryoprotective agents (CPAs) on the exposure, vitrification (VIT), and in vitro culture (IVC) of ovarian tissue with regard to the expression and immunolocalization of aquaporins (AQPs) 3 and 9 in ovine preantral follicles. Tissues were treated as follows: Experiment I: (1) control (without exposure to CPAs), (2) e-EG (exposure to ethylene glycol), (3) er-EG (exposure to and removal of EG), (4) e-DMSO (exposure to dimethyl sulfoxide), (5) er-DMSO (exposure to and removal of DMSO), (6) e-EG+DMSO (exposure to EG+DMSO), (7) er-EG+DMSO (exposure to and removal of EG+DMSO); Experiment II: (1) control, (2) VIT, (3) IVC, (4) VIT-IVC. In Experiment I, following er-EG or er-DMSO, tissue showed the down-regulation (P < 0.05) of AQP3 mRNA. The mRNA transcript levels were reduced (P < 0.05) for AQP9 in tissue following er-EG+DMSO. Immunolocalization was positive for both proteins (AQP3 and AQP9) on ovine preantral follicles following all treatments, except in the e-EG+DMSO group. In Experiment II, the mRNA levels of AQP3 and AQP9 following VIT treatment were similar (P > 0.05) to that of the control group. Nevertheless, VIT-IVC treatment led to the down-regulation of mRNA of AQP3 and AQP9. Thus, AQP3 and AQP9 act in a mutually dependent way, maintaining the cell homeostasis that is essential for the ovary cryopreservation process. Furthermore, the changes in the expression profiles of mRNA and protein after culture are a strong indicator that in vitro conditions have to be strictly controlled to ensure follicle viability and functionality.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil. debissales@gmail.com.
authors.forename: A D
authors.initials: AD
authors.lastname: Sales
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: A B G
authors.initials: AB
authors.lastname: Duarte
authors.affiliation: Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: R R
authors.initials: RR
authors.lastname: Santos
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: K A
authors.initials: KA
authors.lastname: Alves
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: L F
authors.initials: LF
authors.lastname: Lima
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: G Q
authors.initials: GQ
authors.lastname: Rodrigues
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: I R
authors.initials: IR
authors.lastname: Brito
authors.affiliation: Group of Research in Biology of Reproduction - Department of Animal Science, Federal University of Ceará - UFC, Fortaleza, CE, Brazil.
authors.forename: C H
authors.initials: CH
authors.lastname: Lobo
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: J B
authors.initials: JB
authors.lastname: Bruno
authors.affiliation: INRA Physiologie de la Reproduction et des Comportements, 37380, Nouzilly, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Locatelli
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará - UECE, Fortaleza, CE, Brazil.
authors.forename: A P R
authors.initials: AP
authors.lastname: Rodrigues
mesh_terms: D000818:Animals; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D005260:Female; D007150:Immunohistochemistry; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010053:Ovary / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034561:Sheep, Domestic / Q000378:metabolism*; D046509:Tissue Culture Techniques; D058989:Vitrification / Q000187:drug effects
chemical_list: D020346:Aquaporins; D003451:Cryoprotective Agents; D012333:RNA, Messenger
keywords: Aquaporins; Cryoprotective agents; Gene expression; Ovary; Vitrification