FOLLICULOGENESIS: IMMORTALIZED GRANULOSA CELL LINES

描述（改编自申请人的描述）：主要目的 利用转基因技术， 永生化颗粒细胞（GC）系， 促卵泡激素受体（FSHR）。 在试点研究中， 研究人员制备了FSHR基因启动子-SV 40-T抗原cDNA 特异性靶向T抗原（Tag）mRNA表达的构建体 F1代小鼠的性腺。 他们的理由是基于 Windle等（MolEndo 4：597，1990）的转基因方法，其中LH 产生的α-亚单位基因启动子-SV 40-T-抗原cDNA构建体 垂体前叶肿瘤，其中永生LH分泌细胞 线是派生的。 他们建议开发表达FSH受体的GC细胞系 以同样的方式，但已将该方法扩展到F2代 因为GC肿瘤不会在它们的半合子F1后代中发生。的 目标是：1）加速肿瘤在 通过新生DES处理的纯合小鼠;和2）发育和 表征来自标签诱导的和/或DES-标签诱导的GC系 卵巢肿瘤FSHR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。 他们的存在可以让调查人员 在一个扩展的应用程序中，以确定 GC中有丝分裂和凋亡的途径是由FSH唯一调节的。 它们也可能为分子生物学提供极其重要的资源。 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.