Wang X N, Greenwald G S
Department of Physiology, Ralph L. Smith Research Center, University of Kansas Medical Center, Kansas City 66160.
Endocrinology. 1993 May;132(5):2009-16. doi: 10.1210/endo.132.5.8477652.
Adult hypophysectomized FSH-primed mice were used to study ovulation, fertilization, and preimplantation embryo development. Twelve days after hypophysectomy, animals were injected sc with oFSH (4 micrograms/day) twice a day for 3 days. This resulted in large preovulatory follicles that did not secrete estrogen. Concurrent with the last FSH injection, either hCG (5 IU) or human recombinant FSH (10 IU) was injected sc to induce ovulation. Animals were mated or not and killed 1-4 days later. The ovulation rate was similar for both the hCG-induced group (FH) and the FSH-induced group (FF), 97% and 90%, respectively. About 45% of the FH mice mated successfully with 56% of the eggs fertilized compared to only 25% of the FF mice with 45% of the eggs fertilized. However, only 5% of ovulated eggs developed to four-cell stages in vivo by day 3 for the FH animals and none in the FF group. To determine the reasons for the in vivo retarded embryo development, embryos at the one- or two-cell stage were collected on day 2 from the FH group. After 96 h of culture, 22% of two-cell embryos were converted to blastocysts, and 11% of one-cell eggs divided to the four-cell stage. In contrast, 80% of two-cell embryos from normal mice develop into blastocysts by 72 h of culture. The ovarian incubation medium and serum were used to measure progesterone (P4), androstenedione (A), and estradiol (E2). The patterns of serum and in vitro production of steroids were parallel. In FH mice, P4 increased immediately on the day after hCG injection (day 1) and decreased progressively on days 2 and 3; A and E2 levels increased on day 2, A decreased on day 3, and E2 decreased on day 4. When human recombinant FSH was used to induce ovulation, there were no significant changes in serum P4 and A; E2 levels were about 4 times higher on day 1 than in the FSH-primed control, then dropped to baseline levels on days 2 and 3. However, on day 3 in both the FH and FF groups, FSH receptors were still present on the granulosa cells of antral follicles, and LH/hCG receptors were present on the granulosa cells of large antral follicles and newly formed corpora lutea.(ABSTRACT TRUNCATED AT 400 WORDS)
成年垂体切除并用促卵泡激素(FSH)预处理的小鼠被用于研究排卵、受精和植入前胚胎发育。垂体切除12天后,动物每天皮下注射卵泡刺激素(oFSH,4微克/天),每天两次,共3天。这导致形成了不分泌雌激素的大型排卵前卵泡。在最后一次FSH注射的同时,皮下注射人绒毛膜促性腺激素(hCG,5国际单位)或重组人促卵泡激素(10国际单位)以诱导排卵。动物进行交配或不交配,1 - 4天后处死。hCG诱导组(FH)和FSH诱导组(FF)的排卵率相似,分别为97%和90%。与FF组只有25%的小鼠成功交配且45%的卵子受精相比,约45%的FH组小鼠成功交配,56%的卵子受精。然而,到第3天,FH组只有5%的排卵卵在体内发育到四细胞阶段,而FF组没有。为了确定体内胚胎发育迟缓的原因,在第2天从FH组收集一细胞或二细胞阶段的胚胎。培养96小时后,22%的二细胞胚胎发育为囊胚,11%的一细胞卵分裂到四细胞阶段。相比之下,正常小鼠的二细胞胚胎在培养72小时后80%发育为囊胚。使用卵巢培养液和血清来测量孕酮(P4)、雄烯二酮(A)和雌二醇(E2)。血清和体外类固醇生成模式相似。在FH小鼠中,hCG注射后第1天(第1天)血清P4立即升高,在第2天和第3天逐渐下降;A和E2水平在第2天升高,A在第3天下降,E2在第4天下降。当使用重组人促卵泡激素诱导排卵时,血清P4和A无显著变化;E2水平在第1天比FSH预处理对照组高约4倍,然后在第2天和第3天降至基线水平。然而,在第3天,FH组和FF组的窦状卵泡颗粒细胞上仍存在FSH受体,大的窦状卵泡和新形成的黄体的颗粒细胞上存在促黄体生成素/人绒毛膜促性腺激素(LH/hCG)受体。(摘要截断于400字)