Identification of novel ovarian derived oocyte maturation factor and their clinical application

利用DNA微阵列分析，我们发现了一些新的卵母细胞成熟因子的候选物。最终候选体是根据配体在卵巢体细胞中的定位及其在卵母细胞和/或卵丘细胞中的同源受体来选择的。我们可以在化验中找到大约20个候选者。然后，我们对最终候选的卵母细胞核成熟进行了功能分析。用这些配体处理未成熟卵母细胞，研究它们在生发囊泡破裂（GVBD）和第一极体挤压中的作用。结果表明，多种因素可诱发GVBD、第一极体挤压或GVBD和第一极体挤压并存。此外，我们研究了它们对细胞质成熟的贡献，发现有几个因素可以诱导细胞质成熟。在新发现的因子中，我们报道了谷物细胞系衍生的神经营养因子。一些其他因素的报告已提交给期刊，正在审查中。在动物实验基础上发现的新的卵母细胞成熟因子，在人卵母细胞成熟中进一步探讨其表达及其对卵母细胞成熟的影响。分别使用手术治疗患者的卵巢和体外受精- et患者的剩余未受精卵母细胞，通过RT-PCR和免疫组织化学检测配体及其受体的表达。在卵母细胞中发现了新的卵母细胞成熟因子，在卵母细胞中发现了颗粒细胞或卵泡细胞及其受体。此外，我们研究了卵母细胞成熟与人卵泡液中新型卵母细胞成熟因子水平的关系，以及从体外受精-体外受精患者中获得的卵丘细胞。虽然我们可以在关联中发现积极的趋势，但由于样本数量少，它们没有达到显著水平。因此，我们试图为本研究获得更多的样本。因为我们没有患者可以获得完全的知情同意，所以没有对患者的遗传异常进行研究。我们希望将来能找到这样的病人。我们相信我们的研究结果将引领人类新的卵母细胞成熟因子及其调控的发现，并为建立体外成熟卵母细胞的临床应用以及在IVF-ET治疗中获得优质成熟卵母细胞的新方法提供指导。少

Using the DNA microarray analyses, we found some candidates for novel oocyte maturation factors. Final candidates were chosen based on the localization of the ligands in ovarian somatic cells and their cognate receptors in oocytes and/or cumulus cells. We could get approximately 20 candidates in the assay. Then, we have performed functional analyses for oocyte nuclear maturation to the final candidates. Immature oocytes were treated with the ligands to examine their roles in germinal vesicle breakdown (GVBD) and extrusion of first polar body. We found factors could induce GVBD, extrusion of first polar body, or both GVBD and extrusion of first polar body. Furthermore, we examined their contribution to cytoplasmic maturation and found several factors could induce cytoplasmic maturation. Among novel identified factors, we reported grail-cell line derived neurotrophic factor. Some reports for other factors are submitted to journals and are under review. The novel oocyte maturation factors … More identified based on animal studies were further explored in human oocyte maturation by examining their expression and effects on oocyte maturation. Using ovaries and surplus unfertilized oocytes obtained from patients underwent surgical treatment and IVF-ET, respectively, the expression of the ligands and their receptors was determined by RT-PCR and immunohistochemistry. We found the novel oocyte maturation factors in human cumulus and granulosa cells or theca cells and their receptors in oocytes. Furthermore, we examined the association of oocyte maturation with the levels of novel oocyte maturation factors in human follicular fluid and cumulus cells obtained from patients underwent IVF-ET. Although we could find positive tendencies in the association, they were not reached significant levels due to small number of samples. Thus, we try to get more samples for this study. Because we have no patients who could receive full informed consent, studies on genetical abnormality in patients have not performed. We expect to find such patient in future. We believe that our findings lead identification of novel oocyte maturation factors and their regulations in human, and shed a light on the clinical application for establishment of in vitro maturation of oocytes and development of novel methods for obtaining good-quality mature oocytes in IVF-ET treatment. Less