enclosed oocyte, was recorded.To assess meiotic competence, oocytes were removed from follicles, cultured overnight, and the number that underwent germinal vesicle breakdown and reached metaphase II was recorded.Mitochondrial DNA (mtDNA) in individual oocytes was quantified using real-time PCR.Expression of Kit ligand (KL)-1 and -2 by granulosa cells were assayed using real-time RT-PCR.results: Wild-type and mutant mice possessed similar number of primary follicles.However, the mutants possessed fewer secondary and pre-antral follicles than wild-type litter-mates, and no large antral follicles.Oocytes of mutant mice grew to the same size as those of wild-type mice, however, and were able to complete maturation in vitro to metaphase II.Moreover, they contained the same quantity of mtDNA.Expression of KL-1 and KL-2 decreased in the granulosa surrounding fully grown oocytes, consistent with previous reports.However, although FSH can regulate KL expression in vitro, there was no difference between wild-type and mutant oocytes either in the amplitude of the decrease or in the ratio of KL-1:KL-2.conclusion: By the criteria so far examined, oocytes develop normally in vivo in the absence of FSH.Future work will test whether such oocytes can give rise to normal offspring.
BACKGROUNDTo assess the impact of ovarian cystectomy for endometriomas on the ovarian reserve, we evaluated the pre- and post-operative levels of serum anti-Müllerian hormone (AMH). We also analyzed the correlations between factors related to endometriosis and surgery for endometriomas and the serum AMH levels to investigate which factors affect ovarian reserve.
With proper and careful selection of patients, fertility-preserving surgery may be feasible in patients with ovarian malignancies. However, the loss of follicles by oophorectomy and chemotherapy results in decreased ovarian reserve, which consecutively affects reproductive capacity. We evaluated postoperative levels of serum anti-Müllerian hormone (AMH) in women with ovarian malignancies to assess the impact of the fertility-preserving surgery with or without the administration of chemotherapy on ovarian reserve. In 13 patients who underwent the fertility-preserving surgery with (n = 9) or without (n = 4) the administration of chemotherapy, serum AMH levels were measured and compared with serum AMH levels measured in patients undergone cystectomy for benign ovarian tumors as a control. We found that the mean AMH level in the treatment group measured 0.9 ng/mL, which was significantly lower than that measured in the control group (4.70 ± 3.77 ng/mL). The possibility of decreased ovarian reserve occurring in patients with ovarian malignancies following treatment with fertility-preserving surgery with or without the administration of chemotherapy should be considered for fertility planning.
The ovary is a complex endocrine organ responsible for steroidogenesis and folliculogenesis. Follicles consist of oocytes and two primary steroidogenic cell types, the granulosa cells, and the theca cells. Immortalized human granulosa cells are essential for researching the mechanism of steroidogenesis and folliculogenesis. We obtained granulosa cells from a 35-yr-old female and immortalized them by lentivirus-mediated transfer of several genes so as to establish a human nonluteinized granulosa cell line (HGrC1). We subsequently characterized HGrC1 and investigated its steroidogenic performance. HGrC1 expressed enzymes related to steroidogenesis, such as steroidogenic acute regulatory protein, CYP11A, aromatase, and gonadotropin receptors. Stimulation with FSH increased the mRNA levels of aromatase, which consequently induced the aromatization of androstenedione to estradiol. Activin A increased the mRNA levels of the FSH receptor, which were synergistically up-regulated with FSH stimulation. HGrC1 also expressed a series of ligands and receptors belonging to the TGF-β superfamily. A Western blot analysis showed that bone morphogenetic protein (BMP)-4, BMP-6, and BMP-7 phosphorylated small mother against decapentaplegic (Smad)1/5/8, whereas growth differentiation factor-9 phosphorylated Smad2/3. BMP-15 and anti-Müllerian hormone phosphorylated Smad1/5/8 while also weakly phosphorylating Smad2/3. These results indicate that HGrC1 may possess the characteristics of granulosa cells belonging to follicles in the early stage. HGrC1 might also be capable of displaying the growth transition from a gonadotropin-independent status to gonadotropin-dependent one.
