hFSH, T4 e IGF-I no desenvolvimento in vitro de folículos pré- antrais caprinos

• Ano de defesa: 2014
• Autor(a) principal: Caliman, Sanely Lourenço da Costa
• Orientador(a): Costa, Eduardo Paulino da
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Viçosa
• Programa de Pós-Graduação: Doutorado em Medicina Veterinária
• Departamento: Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de
• País: BR
• Palavras-chave em Português: Carpino - Reprodução; Tiroxina; Ovários; Hormônio folículo estimulante
• Palavras-chave em Inglês: Caprine - Reproduction; Thyroxine; Ovaries; Follicle stimulating hormone
• Área do conhecimento CNPq: CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
• Link de acesso: http://locus.ufv.br/handle/123456789/1473

The aim of this study was to evaluate the effect of T4, IGF-I and hFSH, alone or in combination, as well as the culture period (1, 7, 14, 21 and 28 days) on survival, activation and in vitro growth of caprine preantral ovarian follicles (PAOF). For Phase I, II and III, a fragment/follicle will be removed for classic histology and Transmission Electron Microscopy (TEM) constituting the fresh control. Then, the remaining fragments/follicles will be cultured in situ (included in ovarian tissue fragments) for 1 and 7 days (Phase I and II) and 1, 7, 14, 21 and 28 days (Phase III), (at 39°C in a kiln and 5% of CO2) in -MEM+ in the absence or presence of different concentrations of T4 (stage 1), IGF-1 (stage 2) and hFSH (stage 3). In Phase II, the fragments/follicles will be cultured on plates containing -MEM+ supplemented with T4, IGF-I and FSH, in combination at the best concentrations defined in Phase I. In Phase III, the fragments/follicles will be cultured in -MEM+ or in the best medium (media) obtained in Phase II for 1, 7, 14, 21 and 28 days. For the experiments conducted in Phase I, the results showed that after seven days of culture, 10 ng/ml of hFSH is able to promote the activation of primordial follicles and to maintain the ultrastructural integrity of follicles cultured for up to seven days. The addition of 100 ng/ml of IGF-I to the culture medium activate the developing follicles with one day of culture. Furthermore, the use of 20 ng/ml of T4 is effective in maintaining the survival, activation and growth of follicles during seven days of culture. In Phase II, the interaction of T4, hFSH and IGF-I performed during the culture showed that the combination of T4/hFSH (20/10ng/mL) is beneficial for the in vitro development of preantral follicles, during the survival, growth and ultrastructural aspects. The results presented in Phase III demonstrate that the long- duration culture at the concentration of 20/10 ng/ml of T4/hFSH promoted the survival, activation of primordial follicles and follicular growth during seven days of culture. In conclusion, the results of this study showed that the use of hFSH (10ng/ml), IGF-I (100ng/ml) and T4 (20ng/ml) promotes the maintenance of follicle survival and the development of caprine preantral follicles. Moreover, the association of T4 and hFSH xxiiensured the survival of follicles and maintained follicular growth. Follicles cultured for a long duration period (28 days) is necessary for the understanding of the media to be used during the in vitro culture, demonstrating the need for further interactions between substances, for the discovery of a culture medium which allows the development of follicles during folliculogenesis in the preantral and antral stage.