title: Anethole improves the in vitro development of isolated caprine secondary follicles.
abstract: The aim of this study was to investigate the effect of three concentrations of anethole (30, 300, and 2000 μg/mL) on survival, antrum formation, follicular diameter, and oocyte maturation in the caprine species. The study also evaluated the effects of anethole on transcripts of ICAM-1, CAV-1, TIMP-2, and PAI-1 genes and levels of reactive oxygen species (ROS) in isolated goat preantral ovarian follicles before and after in vitro culture for 18 days. Preantral follicles were isolated from goat ovaries and individually cultured in alpha minimum essential medium modified (α-MEM+), defined as the control treatment, α-MEM+supplemented with ascorbic acid at a concentration of 100 μg/mL (AA), or α-MEM+supplemented with three different concentrations of anethole (30, 300, 2000 μg/mL) for a period of 18 days. Treatments were named as α-MEM+, AA, AN30, AN300, and AN2000, respectively. After culture, the follicles were opened, the cumulus oocytes complex (COCs) were removed and matured in vitro. The walls of the follicles were used for the quantitation of mRNA by quantitative real-time polymerase chain reaction. Finally, the medium collected at the end of culture was used for the measurements of ROS. After 18 days of culture, the AA treatment showed the percentage of intact follicles and follicular diameter significantly higher compared with the other treatments. However, daily growth rate, antrum formation, and also oocyte diameter were similar among the treatments. In addition, compared with AA, the rate of oocytes for in vitro maturation (diameter ≥ 110 μm) and the meiosis resumption rate were significantly higher in the treatments AN30 and AN2000, respectively. When assessing gene related to remodeling of the basement membrane, significant differences in mRNA levels for ICAM-1, CAV-1, TIMP-2, and PAI-1 were observed in comparison with Day 0, i.e., in the noncultured control. In addition, the ROS from Day 12, all treatments with the addition of anethole have significantly lower values of ROS than α-MEM+and AA. In conclusion, the addition of anethole to the in vitro culture medium was able to improve the development of goat preantral follicles by reducing concentrations of ROS and increasing the percentage of oocytes able to resume meiosis.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil. Electronic address: naizarcangela@gmail.com.
authors.forename: N A R
authors.initials: NAR
authors.lastname: Sá
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: V R
authors.initials: VR
authors.lastname: Araújo
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: H H V
authors.initials: HHV
authors.lastname: Correia
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: A C A
authors.initials: ACA
authors.lastname: Ferreira
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: D D
authors.initials: DD
authors.lastname: Guerreiro
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: A M
authors.initials: AM
authors.lastname: Sampaio
authors.affiliation: Laboratory of Reproduction Biotechnology, Campus Uruguayana, Federal University of Pampa, Uruguaiana, RS, Brazil.
authors.forename: E
authors.initials: E
authors.lastname: Escobar
authors.affiliation: Laboratory of Reproduction Biotechnology, Campus Uruguayana, Federal University of Pampa, Uruguaiana, RS, Brazil.
authors.forename: F W
authors.initials: FW
authors.lastname: Santos
authors.affiliation: Department of Zootecnia, Federal University of Ceará, Fortaleza, CE, Brazil.
authors.forename: A A
authors.initials: AA
authors.lastname: Moura
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: C H
authors.initials: CH
authors.lastname: Lôbo
authors.affiliation: Laboratory of Electrophysiology, Superior Institute of Biomedical Sciences, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: V M
authors.initials: VM
authors.lastname: Ceccatto
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: C C
authors.initials: CC
authors.lastname: Campello
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: A P R
authors.initials: APR
authors.lastname: Rodrigues
authors.affiliation: Laboratory of Electrophysiology, Superior Institute of Biomedical Sciences, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: J H
authors.initials: JH
authors.lastname: Leal-Cardoso
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
mesh_terms: D000081248:Allylbenzene Derivatives; D000818:Animals; D000840:Anisoles / Q000494:pharmacology*; D005260:Female; D006041:Goats / Q000502:physiology*; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods / Q000662:veterinary*; D006080:Ovarian Follicle / Q000254:growth & development*; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000081248:Allylbenzene Derivatives; D000840:Anisoles; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; C006578:anethole
keywords: Anethole; Antioxidant; In vitro culture; In vitro maturation; Reactive oxygen species