M K B Serafim1, A B G Duarte1, G M Silva1, C E A Souza2, D M Magalhães-Padilha3, A A A Moura1, L D M Silva4, C C Campello1, J R Figueiredo1. 1. Faculty of Veterinary Medicine, Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Veterinary Science Post Graduation Program (VSPGP), State University of Ceara, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60740-000, Brazil. 2. Laboratory of Animal Physiology, Department of Animal Science, Federal University of Ceara, Av. Mister Hull S/N, Campus Pici, Fortaleza, CE CEP: 60021-970, Brazil. 3. Potiguar University, Laureate International Universities, Av. Senador Salgado Filho, 1610, Lagoa Nova, Natal, RN CEP: 59056-000, Brazil. Electronic address: dmmvet@hotmail.com. 4. Faculty of Veterinary Medicine, Laboratory of Carnivore Reproduction (LRC), State University of Ceara, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE CEP: 60740-000, Brazil.
Abstract
OBJECTIVE: Evaluate the effect of different concentrations of growth hormone (GH) on the in vitro development of domestic dog (Canis lupus familiaris) preantral follicles in the presence or absence of follicle stimulating hormone (FSH). METHODS: Secondary preantral follicles, isolated by microdissection, were cultured in a medium composed of αMEM with bovine serum albumin (BSA), glutamine, hypoxanthine, insulin, transferrin, selenium and ascorbic acid (αMEM(+)-control) added at different concentrations of GH (GH10 ng/ml or GH50 ng/ml) and FSH (GH10+FSH, GH50+FSH). Follicle development was evaluated based on the percentage of intact follicles, antrum formation, follicular diameter, follicular viability using fluorescent markers and estradiol production. RESULTS: GH50 was the only treatment that maintained the same percentage of normal morphologically follicles from day 0 to day 18 of culture (P<0.05). For all treatments, except the control, follicles were viable throughout the 18 days of culture (P<0.05). GH50 supplemented with FSH (GH50+FSH) resulted in the highest average follicular diameter (P<0.05) from day 12 to 18. Follicles from both the control and the GH50+FSH treatment groups actively and increasingly secreted estradiol from day 6 to 18 of culture (P<0.05). CONCLUSIONS: Our study demonstrates that GH benefits the maintenance of follicular morphology in a dose-dependent manner and, in association with FSH, stimulates in vitro follicular growth and estradiol production.
OBJECTIVE: Evaluate the effect of different concentrations of growth hormone (GH) on the in vitro development of domestic dog (Canis lupus familiaris) preantral follicles in the presence or absence of follicle stimulating hormone (FSH). METHODS: Secondary preantral follicles, isolated by microdissection, were cultured in a medium composed of αMEM with bovineserum albumin (BSA), glutamine, hypoxanthine, insulin, transferrin, selenium and ascorbic acid (αMEM(+)-control) added at different concentrations of GH (GH10 ng/ml or GH50 ng/ml) and FSH (GH10+FSH, GH50+FSH). Follicle development was evaluated based on the percentage of intact follicles, antrum formation, follicular diameter, follicular viability using fluorescent markers and estradiol production. RESULTS:GH50 was the only treatment that maintained the same percentage of normal morphologically follicles from day 0 to day 18 of culture (P<0.05). For all treatments, except the control, follicles were viable throughout the 18 days of culture (P<0.05). GH50 supplemented with FSH (GH50+FSH) resulted in the highest average follicular diameter (P<0.05) from day 12 to 18. Follicles from both the control and the GH50+FSH treatment groups actively and increasingly secreted estradiol from day 6 to 18 of culture (P<0.05). CONCLUSIONS: Our study demonstrates that GH benefits the maintenance of follicular morphology in a dose-dependent manner and, in association with FSH, stimulates in vitro follicular growth and estradiol production.
Authors: Emina Ipsa; Vinicius F Cruzat; Jackob N Kagize; John L Yovich; Kevin N Keane Journal: Front Endocrinol (Lausanne) Date: 2019-11-12 Impact factor: 5.555