title: Balance of insulin and FSH concentrations improves the in vitro development of isolated goat preantral follicles in medium containing GH.
abstract: The aim of this study was to evaluate the effect of different combinations of insulin and FSH concentrations in culture media containing GH on the in vitro follicle morphology, antrum formation, growth rates, estradiol (E2) production, oocyte viability and maturation as well as gene expression for FSHR, GHR, INSR, CYP19A1, CYP17, 3ßHSD. Secondary follicles were individually cultured for 18 days in a basic medium containing 50ng/mL GH supplemented with low insulin concentration (INS-LW: 10ng/mL) or high insulin concentration (INS-HG: 10μg/mL) alone or with a fixed FSH concentration (FSH100: 100ng/mL) or with increasing FSH concentrations (FSH-SEQ: 100ng/mL, days 0-6; 500ng/mL, days 6-12; 1000ng/mL days 12-18). In the INS-LW treatment was observed a higher (P<0.05) incidence of normal follicles at day 18 of culture. However, overall higher (P<0.05) follicular growth, oocyte diameter and meiotic resumption rates were obtained using INS-HG+FSH 100. The INS-HG and INS-HG+FSH100 treatments showed higher E2 production and mRNA levels for CYP19A1, CYP17, 3βHSD when compared to INS-LW and INS-LW+FSH100. However, the addition of increasing FSH concentration, regardless of insulin concentration, did not improve the follicular growth, meotic resumption, E2 production or gene expression of steroidogenic enzymes when compared with INS-HG+FSH100. In conclusion, in presence of GH, a basic medium supplemented with 10μg/mL insulin and 100μg/mL FSH throughout the culture period, improves follicular and oocyte growth, oocyte meiotic resumption and E2 production from isolated preantral caprine follicles cultured in vitro.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary, State University of Ceará, Fortaleza, CE, Brazil. Electronic address: annaclaraaccioly@yahoo.com.br.
authors.forename: A C A
authors.initials: ACA
authors.lastname: Ferreira
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: C
authors.initials: C
authors.lastname: Maside
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: N A R
authors.initials: NAR
authors.lastname: Sá
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary, 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, 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, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: J
authors.initials: J
authors.lastname: Leiva-Revilla
authors.affiliation: Laboratory of Animal Physiology, Department of Animal Science, Federal University of Ceará, 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, State University of Ceará, Fortaleza, CE, Brazil; Health Sciences Center, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: V R
authors.initials: VR
authors.lastname: Araújo
authors.affiliation: Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, USA.
authors.forename: G A
authors.initials: GA
authors.lastname: Apgar
authors.affiliation: Department of Animal Reproduction, Faculty of Veterinary, Federal University Fluminense, Rio de Janeiro, RJ, Brazil.
authors.forename: F Z
authors.initials: FZ
authors.lastname: Brandão
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary, State University of Ceará, 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, State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: C C
authors.initials: CC
authors.lastname: Campello
mesh_terms: D000818:Animals; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004967:Estrogens / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000008:administration & dosage / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D006041:Goats / Q000502:physiology*; D013006:Growth Hormone / Q000494:pharmacology*; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods; D007328:Insulin / Q000008:administration & dosage / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D046509:Tissue Culture Techniques / Q000662:veterinary
chemical_list: D003470:Culture Media; D018076:DNA, Complementary; D004967:Estrogens; D007328:Insulin; D012313:RNA; D005640:Follicle Stimulating Hormone; D013006:Growth Hormone
keywords: FSH; Follicle development; Goat; Insulin; Oocyte maturation