Dode M A N, Graves C N
Animal Reproduction Laboratory, Embrapa Genetic Resources and Biotechnology, Parque Estação Biológica, Avenue W5 Final Norte, Brasília CEP 70770-900, DF, Brazil.
Anim Reprod Sci. 2003 Sep 15;78(1-2):99-110. doi: 10.1016/s0378-4320(03)00080-0.
The role of estradiol-17beta on nuclear and cytoplasmic maturation of pig oocytes was investigated in the present study. To determine the estradiol effect, oocytes were cultured for 42 h in a steroid free medium composed of mTCM-199 supplemented with LH, FSH and 10% charcoal extracted follicular fluid. Estradiol receptor (ER), detected by a binding assay, were present in cumulus cells and oocytes during maturation with higher levels observed at 24 h of culture in the oocytes and at 36 h in the cumulus cells. To block estradiol action an antiestrogen (1-p-dimethylaminoethoxyphenyl-1,2-diphenyl-1-butene (tamoxifen)) was added to the maturation medium at various concentrations. The percentage of treated oocytes that underwent nuclear maturation was similar (P>0.05) to the control group. Cytoplasmic maturation, determined by the ability to form female pronucleus (FPN) and male pronucleus (MPN), was not different (P>0.05) among all groups. The presence of 4-hydroxy-4-androstene-3-17-dione (4-OHA) also did not influence nuclear (P>0.05) or cytoplasmic maturation (P>0.05). The results suggest that estradiol is not involved in maturation of pig oocytes. However, the present experiment used pronuclei formation as the endpoint, no studies were done in regard to estradiol's effects on the embryonic development.
本研究调查了17β-雌二醇对猪卵母细胞核成熟和细胞质成熟的作用。为确定雌二醇的作用,将卵母细胞在不含类固醇的培养基中培养42小时,该培养基由添加了促黄体生成素(LH)、促卵泡激素(FSH)和10%经活性炭处理的卵泡液的改良组织培养液-199(mTCM-199)组成。通过结合试验检测发现,在成熟过程中,卵丘细胞和卵母细胞中均存在雌二醇受体(ER),培养24小时时在卵母细胞中观察到较高水平,培养36小时时在卵丘细胞中观察到较高水平。为阻断雌二醇的作用,将抗雌激素药物(1-对二甲氨基乙氧基苯基-1,2-二苯基-1-丁烯(他莫昔芬))以不同浓度添加到成熟培养基中。接受核成熟处理的卵母细胞百分比与对照组相似(P>0.05)。通过形成雌原核(FPN)和雄原核(MPN)的能力来确定的细胞质成熟,在所有组中没有差异(P>0.05)。4-羟基-4-雄烯-3,17-二酮(4-OHA)的存在也不影响核成熟(P>0.05)或细胞质成熟(P>0.05)。结果表明,雌二醇不参与猪卵母细胞的成熟。然而,本实验以原核形成为终点,未对雌二醇对胚胎发育的影响进行研究。