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者。基于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选择最终候选物。我们可以在检测中得到大约20个候选人。然后，我们对最终候选人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，研究它们在生殖囊泡破裂（GVBD）和第一极体排出中的作用。我们发现有多种因素可以诱发GVBD、第一极体排出或GVBD和第一极体排出。此外，我们研究了它们对细胞质成熟的贡献，发现几种因素可以诱导细胞质成熟。在新发现的因子中，我们报道了粒细胞源性神经营养因子。其他因素的一些报告已提交给期刊，正在审查中。新的卵母细胞成熟因子 ...更多信息 通过检测它们的表达和对卵母细胞成熟的影响，进一步探索了基于动物研究鉴定的它们在人卵母细胞成熟中的作用。分别采用手术治疗和IVF-ET患者的卵巢和剩余未受精卵母细胞，通过RT-PCR和免疫组织化学方法测定配体及其受体的表达。我们在人卵丘和颗粒细胞或卵泡膜细胞中发现了新的卵母细胞成熟因子及其受体。此外，我们研究了从接受IVF-ET的患者中获得的人卵泡液和卵丘细胞中卵母细胞成熟与新型卵母细胞成熟因子水平的关系。虽然我们可以发现积极的趋势，在协会，他们没有达到显着水平，由于样本数量少。因此，我们试图为这项研究获得更多的样本。由于我们没有患者可以获得完全知情同意，因此尚未对患者进行遗传异常研究。我们希望将来能找到这样的病人。我们相信，我们的研究结果导致识别新的卵母细胞成熟因子及其调控在人类，并为建立卵母细胞体外成熟的临床应用和开发新的方法，以获得高质量的成熟卵母细胞在IVF-ET治疗的启示。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