RESUMO
Epidermal growth factor (EGF) has been considered a potential regulator of meiotic and cytoplasmic maturation in mammalian oocytes, but inconsistencies exist between earlier studies, probably due to differences in the culture conditions used. Using a serum- and hormone-free in vitro maturation (IVM) medium, this study investigated the specific contribution of EGF on IVM of porcine (Sus scrofa) oocytes and its interactive effects with follicle-stimulating hormone (FSH), porcine follicular fluid (pFF), cumulus cells, and serum. It was noteworthy that EGF functionally mimicked the action of FSH and could completely replace FSH for nuclear maturation (83.2+/-4.4% vs. 55.9+/-5.2%; mean+/-SEM), whereas EGF had a synergistic effect with FSH on cytoplasmic maturation of porcine oocytes (P<0.05). Specific inhibition of EGF receptor (EGFR) by tyrphostin AG 1478 inhibited both EGF- and FSH-induced meiotic resumption (17.9+/-5.2% and 18.2+/-4.4%, respectively), thereby suggesting that EGFR signaling pathway was essential for oocyte reentry into the meiotic cell cycle. Furthermore, it is possible that FSH action occurs via the EGFR signaling pathway to induce meiotic maturation, although alternate pathways could not be excluded. There were also individual or combined effects of cumulus cells, FSH, serum, and pFF with EGF on IVM of porcine oocytes (P<0.05). Although FSH had a synergistic effect with EGF on cytoplasmic maturation, pFF masked the effects of EGF on both nuclear and cytoplasmic maturation of porcine oocytes (P<0.05). Moreover, the presence of cumulus cells was essential for EGF action. In conclusion, a defined system was used to better examine the effects of EGF. We inferred that EGF functionally mimics FSH for nuclear maturation of porcine oocytes, and its exogenous supplementation into IVM medium can optimize the beneficial effects of FSH on cytoplasmic maturation of oocytes to obtain enhanced embryo development in vitro.
Assuntos
Núcleo Celular/efeitos dos fármacos , Fator de Crescimento Epidérmico/farmacologia , Hormônio Foliculoestimulante/farmacologia , Oócitos/efeitos dos fármacos , Suínos , Animais , Células Cultivadas , Interações Medicamentosas , Técnicas de Cultura Embrionária , Embrião de Mamíferos/citologia , Embrião de Mamíferos/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Feminino , Fertilização in vitro/métodos , Fertilização in vitro/veterinária , Oócitos/metabolismo , Oócitos/ultraestrutura , Oogênese/efeitos dos fármacos , Biossíntese de Proteínas/efeitos dos fármacos , Quinazolinas , Suínos/embriologia , Equivalência Terapêutica , Tirfostinas/farmacologiaRESUMO
To investigate the effects of irradiation on structural and functional properties of egg white proteins, which enhance foaming ability, egg white was separated and irradiated at doses of 0, 2.5, and 5 kGy. The foaming ability of egg white was increased, whereas foam stability was decreased by irradiation. Turbidity and protein oxidation of egg white was increased by irradiation with an increase of irradiation dose. The content of free sulfhydryl and disulfide was not affected by irradiation. According to 2-dimensional electrophoresis analysis, it was demonstrated that protein scissions are the main changes caused by irradiation and this protein modification may be the main reason for the improvement in foaming ability of egg white.