THE ROLE OF PATCHED 1 AND SUPPRESSOR OF FUSED IN THE OVARY

PATCHED 1 和 FUSED 抑制剂在卵巢中的作用

• 批准号: 7959953
• 负责人: ZI-JIAN LAN
• 金额: $ 27.8万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959953
• 关键词: Affect; Alleles; Automobile Driving; Cell Differentiation process; Cell Proliferation; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Cytochrome P450; DNA; Defect; Development; Diagnosis; Dysplasia; Embryo; Endocrine; Epidermal Growth Factor; Erinaceidae; Family; Female; Female infertility; Female sterility; Fertility; Funding; Germ Cells; Goals; Gonadotropins; Grant; Growth; Growth Differentiation Factor 9; Growth Factor; Infertility; Institution; Knowledge; Lead; Mammals; Mesenchymal; Mixed Function Oxygenases; Molecular; Mus; Mutant Strains Mice; Mutation; Oocytes; Oogenesis; Ovarian; Ovary; Phenotype; Play; Process; Production; Reproduction; Research; Research Personnel; Resources; Response Elements; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Somatomedins; Source; Sterility; Steroids; Theca-Lutein Cells; Transforming Growth Factors; United States National Institutes of Health; base; cell type; folliculogenesis; intercellular communication; intraovarian; member; mouse model; mutant; postnatal; promoter; smoothened signaling pathway; theca cell

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Folliculogenesis and oogenesis are complex processes which occur in the ovary and result in the production of fertilizable female germ cells. These processes require proper intercellular communication between many cell types including oocyte, granulosa, theca, and luteal cells within the ovary. Many intraovarian growth factors and endocrine factors such as insulin-like growth factor, epidermal growth factor, steroids, members of the transforming growth factor b family, the Wnt/Frizzled family, and gonadotropins are involved in these processes, affecting female reproduction. Here we propose that the hedgehog (Hh) signaling pathway, one of the major signaling pathways in driving cell proliferation and directing cell differentiation during embryonic and postnatal development, will play a role in folliculogenesis and oogenesis to regulate female fertility in mammals. There are many components of the Hh signaling pathway. In this proposal, we will focus on characterization of the roles of patched-1 (Ptc) and suppressor of fused (Sufu), two negative regulators of Hh signaling, in the ovary. Ptc is expressed in oocytes and theca cells in mouse ovary. Mice containing a natural occurring Ptc mutation (Ptcmes) display female sterility. Sufu is also highly expressed in mouse oocytes. Putative Gli DNA response elements are present in the promoters of Growth Differentiation Factor-9 (GDF-9) and cytochrome P450 17alpha-hydroxylase (Cyp17), which are known to play an important role in normal ovarian functions. Based on this information, we hypothesized that Hh signaling molecules Ptc and Sufu will play a role in folliculogenesis and oogenesis likely by regulating GDF-9 and Cyp17 expression in oocytes or thecal cells, affecting female fertility in mammals. Genetically modified Ptc and Sufu mouse models will be used to determine their ovarian roles in this study. The specific aims are to (1) Characterize the ovarian phenotypes of sterile Ptc mutation mice carrying a spontaneous mesenchymal dysplasia (mes) mutation (Ptcmes/mes); and (2) Investigate the synergistic effects of Ptc and Sufu in the ovaries from Sufu+/-;Ptcmes/mes mutant mice. Fertility analysis in combination with morphological, histological, and molecular analyses will be performed to characterize ovarian phenotypes of these Sufu and Ptc mutant mice. It is expected that deletion of Ptc and/or Sufu will mimic ectopic Hh signaling in oocytes and/or thecal cells to stimulate their growth or proliferation, result in aberrant folliculogenesis and oogenesis, and eventually lead to female infertility. Introduction of a mutant Sufu allele into Ptcmes/mes mice will cause more severe ovarian phenotypes than Ptcmes/mes alone. The goal of this study is to explore whether Hh signaling is functional in the ovary. This study would substantially advance our knowledge about ovarian function and female fertility. Knowledge gained from this study could be used in diagnosis and treatment of female idiopathic infertility.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 卵泡发生和卵子发生是发生在卵巢中的复杂过程，导致可受精的雌性生殖细胞的产生。 这些过程需要在卵巢内的许多细胞类型包括卵母细胞、颗粒细胞、卵泡膜细胞和黄体细胞之间进行适当的细胞间通讯。许多卵巢内生长因子和内分泌因子如胰岛素样生长因子、表皮生长因子、类固醇、转化生长因子B家族成员、Wnt/Frizzled家族和促性腺激素参与这些过程，影响雌性生殖。在这里，我们提出，刺猬（Hh）信号通路，在驱动细胞增殖和指导细胞分化在胚胎和出生后发育的主要信号通路之一，将发挥作用，在卵泡和卵子发生，以调节女性生育哺乳动物。Hh信号通路有许多组成部分。在这个建议中，我们将集中在特性的作用patched-1（Ptc）和抑制融合（Sufu），Hh信号的两个负调节器，在卵巢中。Ptc在小鼠卵巢的卵母细胞和卵泡膜细胞中表达。含有天然存在的Ptc突变（Ptcmes）的小鼠显示雌性不育。腐乳在小鼠卵母细胞中也高度表达。 假定的Gli DNA响应元件存在于生长分化因子-9（GDF-9）和细胞色素P450 17 α-羟化酶（Cyp 17）的启动子中，已知其在正常卵巢功能中起重要作用。基于这些信息，我们假设Hh信号分子Ptc和Sufu可能通过调节卵母细胞或卵泡膜细胞中GDF-9和Cyp 17的表达，在卵泡发生和卵子发生中发挥作用，从而影响哺乳动物的雌性生育力。转基因Ptc和Sufu小鼠模型将用于确定其在本研究中的卵巢作用。具体目的是（1）表征携带自发性间质发育不良（mes）突变（Ptcmes/mes）的不育Ptc突变小鼠的卵巢表型;（2）研究Ptc和Sufu在Sufu+/-;Ptcmes/mes突变小鼠卵巢中的协同作用。将结合形态学、组织学和分子分析进行生育力分析，以表征这些Sufu和Ptc突变小鼠的卵巢表型。预期Ptc和/或Sufu的缺失将模拟卵母细胞和/或卵泡膜细胞中的异位Hh信号传导以刺激它们的生长或增殖，导致异常的卵泡发生和卵子发生，并最终导致女性不育。将突变的Sufu等位基因引入Ptcmes/mes小鼠将导致比单独Ptcmes/mes更严重的卵巢表型。本研究的目的是探讨Hh信号是否在卵巢中起作用。这项研究将大大提高我们对卵巢功能和女性生育能力的认识。本研究为女性特发性不孕症的诊断和治疗提供了依据。