title: Caprine ovarian follicle requirements differ between preantral and early antral stages after IVC in medium supplemented with GH and VEGF alone or in combination.
abstract: The aim of the present study was to evaluate the effect of growth hormone (GH) and vascular endothelial growth factor (VEGF) added alone, sequentially or in combination, in the presence of insulin at physiological concentration (10 ng/mL) on the IVC of two different follicular categories: preantral (experiment 1; Exp.1) and early antral (experiment 2; Exp.2). Isolated follicles were individually cultured for 24 (Exp.1) and 18 days (Exp.2) in the following treatments: αMEM+(Control), or Control medium supplemented with 50 ng/mL GH (GH), 100 ng/mL VEGF (VEGF), the combination of both (GH + VEGF), GH during the first 12 days and VEGF from Day 12 until the end of the culture (GH/VEGF) and vice versa (VEGF/GH). At the end of the culture, cumulus-oocyte complexes from in vitro-grown follicles were recovered and subjected to IVM. The following end points were evaluated: Follicle morphology, growth rates and antrum formation, production of estradiol, progesterone and testosterone, oocyte viability and meiotic stage, as well as relative expression of LHR, Amh, HAS2, PTGS2, CYP17, CYP19A1, and 3βHSD. A considerable amount of viable fully grown oocytes were recovered after the IVC of early antral follicles in all treatments. Nevertheless, the GH treatment presented the highest percentage of fully grown oocytes (60%), mean oocyte diameter (117.74 ± 2.61 μm), and meiotic resumption (50%). Furthermore, GH treatment produced higher (P < 0.05) rates of metaphase II oocytes than all the other treatments, and similar LHR, Amh, and PTGS2 transcript levels to in vivo. Contrary to early antral follicles, preantral follicles were not affected by medium supplementation. In conclusion, the addition of GH to a culture medium containing physiological concentrations of insulin improves oocyte growth and maturation after the IVC of goat early antral follicles.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: J
authors.initials: J
authors.lastname: Cadenas
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: J
authors.initials: J
authors.lastname: Leiva-Revilla
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: L A
authors.initials: LA
authors.lastname: Vieira
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: L B
authors.initials: LB
authors.lastname: Apolloni
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: F L N
authors.initials: FLN
authors.lastname: Aguiar
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: B G
authors.initials: BG
authors.lastname: Alves
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 (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: A P R
authors.initials: APR
authors.lastname: Rodrigues
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: Follicle Biology Laboratory, Center for Reproductive Medicine, UZ Brussel, Brussels, Belgium.
authors.forename: J
authors.initials: J
authors.lastname: Smitz
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil. Electronic address: jrf.lamofopapapers@gmail.com.
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.
authors.forename: C
authors.initials: C
authors.lastname: Maside
mesh_terms: D000818:Animals; D003470:Culture Media; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D006041:Goats; D013006:Growth Hormone / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000502:physiology*; D011374:Progesterone / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000379:methods / Q000662:veterinary*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*
chemical_list: D003470:Culture Media; D042461:Vascular Endothelial Growth Factor A; D013739:Testosterone; D011374:Progesterone; D004958:Estradiol; D013006:Growth Hormone
keywords: Growth hormone; Insulin; Oocyte maturation; Ovarian follicle; Vascular endothelial growth factor