FOLLICULOGENESIS: IMMORTALIZED GRANULOSA CELL LINES

描述(改编自申请者的描述)：主要目标 这个项目的目的是利用转基因技术来开发 永生化颗粒细胞(GC)系，将保持功能 卵泡刺激素受体(FSHR)。在试点研究中， 研究人员制备了FSHR基因启动子-SV40-T抗原cDNA 特异性靶向T抗原(Tag)mRNA表达的构建 对F1小鼠性腺的影响。他们的理由是基于 Windle等人的转基因方法(Mol Endo 4：597,1990) α-亚基基因启动子-SV40-T抗原的构建 垂体前叶肿瘤，从中获得永生化的黄体生成素分泌细胞 获得了线条。他们建议培育表达FSHR的GC系 以同样的方式，但已将方法扩展到F2代 因为GC肿瘤不是在他们半合子的F1后代中发生的。这个 目标将是：1)加速肿瘤的发展 通过新生儿DES治疗的纯合子小鼠；以及2)发育和 标签诱导和/或DES标签诱导的GC品系的特征 卵巢肿瘤中FSHR的表达和反应性。FSHR- 表达细胞将为研究卵泡发生提供强有力的工具 在分子水平上。他们的可用性可能会让调查人员 在一个扩展的应用中，用来确定 Gc中的有丝分裂和凋亡通路受FSH的独特调控。 它们也可能为分子生物学提供至关重要的资源。 卵巢细胞功能的生物学研究。

DESCRIPTION (Adapted from Applicant's Description): The major objective of this project is to utilize transgenic technology to develop immortalized granulosa cell (GC) lines which will maintain functional Follicle-stimulating hormone receptor (FSHR). In pilot studies, the investigators prepared a FSHR gene-promoter-SV40-T-antigen cDNA construct which targeted T-antigen (Tag) mRNA expression specifically to the gonads of the F1 mice. Their rationale is based upon the transgenic approach of Windle et al (Mol Endo 4:597,1990) in which LH alpha-subunit gene promoter-SV40-T-antigen cDNA constructs produced anterior pituitary tumors, from which immortalized LH-secreting cell lines were derived. They propose to develop FSHR-expressing GC lines in the same manner, but have extended the approach to the F2 generation because GC tumors did not develop in their hemizygous F1 offspring. The objectives will be to: 1) accelerate the development of tumors in the homozygous mice by neonatal DES treatment; and 2) develop and characterize GC lines from the Tag-induced and/or DES-Tag-induced ovarian tumors in terms of FSHR expression and responsiveness. FSHR- expressing cells will provide a powerful tool to study folliculogenesis at the molecular level. Their availability may allow the investigators in an expanded application to determine the mechanism(s) by which the pathways of mitosis and apoptosis in GC are uniquely regulated by FSH. They may also provide a critically important resource for molecular biological studies of ovarian cellular function.