DEVELOPMENT OF MAMMALIAN OOCYTE-GRANULOSA CELL COMPLEX

哺乳动物卵母细胞-颗粒细胞复合体的发育

• 批准号: 2198966
• 负责人: JOHN J EPPIG
• 金额: $ 14.77万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-03 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2198966
• 关键词: biomarker; cell aggregation; cell cell interaction; cell differentiation; cell growth regulation; egg /ovum; follicle stimulating hormone; gene expression; granulosa cell; hormone receptor; hormone regulation /control mechanism; hyaluronate; in situ hybridization; laboratory mouse; luteinizing hormone; messenger RNA; molecular cloning; northern blottings; nucleic acid probes; ovariectomy; paracrine; phenotype; secretion; tissue /cell culture

The overall objective of this project is to determine what role the oocyte plays in the fundamental organization, differentiation and function of the ovarian granulosa cells. Granulosa cells of large antral follicles are heterogeneous in their expression of specific phenotypic characteristics. One population of granulosa cells, the mural granulosa cells, is associated with the basal lamina and the other, the cumulus cells, with the oocyte. Such microenvironmental associations suggest that the oocyte and the basal lamina both influence the fate and function of the cells associated with them. Although many factors originating in the pituitary gland, the follicular theca or even in the granulosa cells themselves participate in granulosa cell development and function, the overall goal of this project is to determine what role the oocyte plays in the development and function of the granulosa cells. The hypothesis underlying this work is that the selection of the specific pathway for development, as functional cumulus cells or as mural granulosa cells, is determined by the association of the granulosa cells with either the oocyte or the basal lamina. Furthermore, it is hypothesized that the oocyte is actually the dominant determinative factor in granulosa cell development. More specifically, the default program of granulosa cell differentiation is proposed to yield the mural granulosa cell phenotype, which is augmented by contact with basal lamina, while paracrine factors from the oocyte suppress the expression of this phenotype and promote the cumulus cell phenotype. The following specific aims will address this hypothesis. First, mRNA species expressed specifically by mural and cumulus granulosa cells will be identified and cloned. These will be used as markers of the cumulus and mural granulosa cell phenotypes. Specific Aims 2 and 3 will compare the ability of oocyte secretions and components of basal lamina to affect these phenotypes. It will be determined whether the development of the phenotypic characteristics of mural granulosa cells is suppressed by paracrine factors from oocytes even when the expression of the mural granulosa cell phenotype is promoted by components of basal lamina (Aim 2). Then, it will be determined whether microsurgical removal of the oocyte from oocyte-cumulus cell complexes promotes the expression of the mural granulosa cell phenotype and the loss of the cumulus cell phenotype. If so, it will be determined whether paracrine factors from oocytes maintain the cumulus cell phenotype and suppress the expression of the mural granulosa cell phenotype even when oocytectomized complexes are maintained in contact with basal lamina in vitro (Aim 3). Completion of these aims will further the understanding of how the oocyte influences the fundamental organization of the follicle. This may be by a dominant influence of the oocyte in creating the heterogeneity in the pattern of gene expression and cellular function of granulosa cells. These new perspectives on follicular development could lead to novel approaches to the regulation of fertility and the resolution of ovarian dysfunction by means that target oocyte-somatic cell interactions.

该项目的总体目的是确定卵母细胞的作用 在基本组织，差异化和功能中发挥作用 卵巢颗粒细胞。 大鼻腔卵泡的颗粒细胞是 它们表达特定表型特征的异质。 一个颗粒细胞的群，壁画颗粒细胞是 与基底层和另一个相关 卵母细胞。这种微环境关联表明卵母细胞 基础薄片都影响细胞的命运和功能 与它们相关。尽管许多因素起源于垂体 腺体，卵泡theca，甚至在颗粒细胞中 参与颗粒细胞的开发和功能，总体目标 这个项目的是确定卵母细胞在 颗粒细胞的开发和功能。基础的假设 这项工作是选择特定发展途径， 作为功​​能性积云细胞或壁画颗粒细胞，由 颗粒细胞与卵母细胞或基础的关联 薄片。此外，假设卵母细胞实际上是 颗粒细胞发育中的主要决定因素。更多的 具体而言，颗粒细胞分化的默认程序是 提议生产壁画颗粒细胞表型，该表型已增强 通过与基底层接触，而卵母细胞的旁分泌因子 抑制这种表型的表达并促进积云细胞 表型。以下具体目标将解决这一假设。 首先，由壁画和积云颗粒特别表达的mRNA物种 细胞将被识别和克隆。 这些将用作标记 积云和壁画颗粒细胞表型。 具体目标2和3将 将卵母细胞分泌物和基底层分泌物的能力与 影响这些表型。 将确定是否发展 壁画细胞的表型特征被抑制 即使壁画的表达，卵母细胞的旁分泌因子 颗粒细胞表型是由基底层的成分促进的（AIM 2）。然后，将确定是否要去除显微外科 来自卵母细胞 - 细胞复合物的卵母细胞促进了 壁画颗粒细胞的表型和积云细胞表型的丧失。 如果是这样，将确定是否来自卵母细胞的旁分泌因子 保持积云细胞表型并抑制 即使卵母细胞切除络合物是壁画颗粒细胞表型 维持在体外与基底层接触（AIM 3）。 这些目标的完成将进一步了解卵母细胞 影响卵泡的基本组织。这可能是 卵母细胞在创造异质性中的主要影响 颗粒细胞的基因表达和细胞功能的模式。这些 关于卵泡发展的新观点可能导致新的方法 调节生育能力和卵巢功能障碍的解决 通过靶向卵母细胞的相互作用。