SF1 & CREB: GONADAL ROLES BY A NEW TRANSGENIC APPROACH

支持细胞和颗粒细胞是卵巢的重要体细胞成分。 性腺。这些细胞通过提供 必需的营养素和调节信号，支持其 成熟。反过来，支持细胞和颗粒细胞受到一种 细胞联合的宿主和错综复杂的内分泌和 旁分泌信号。这一系统的复杂性凸显了 需要建立动物模型来研究Sertoli和颗粒如何 细胞因子影响性腺功能。这样的研究最终将 改进节育和性腺治疗方法 畸形和不孕。 SF-1和CREB作为支持细胞和颗粒细胞的关键蛋白质出现 功能。这些蛋白质，像许多其他蛋白质一样，已经被表征 在这些细胞中，但它们在调节生殖细胞发育中的作用 仍然鲜为人知。这项提议测试了这些蛋白质的重要性， 同时开发一种新的转基因方法，特别是将超过- 在支持细胞和颗粒细胞中表达蛋白质。这项工作不会 仅提供有价值的动物模型来阐明性腺功能 SF-1和CREB，但将产生重要的技术优势 对于许多人来说，因为它提供了一种方法来创建表达 蛋白质水平升高，同时使用弱但高度特异的 推动者。 将创造四个转基因小鼠品系。两个将包含 表达Cre重组酶基因的转基因 卵泡刺激素受体启动子。这个启动子是活性的 仅在性腺的支持细胞和颗粒细胞中，因此CRE 重组酶将只存在于转基因小鼠的这些细胞中。二 其他小鼠品系将包含靶向转基因，旨在过度 表达SF-1或CREB的显性否定形式 无所不在的推动者。这些蛋白质的基因将不会被激活 除非发生由CRE促成的重组事件 表达Cre的小鼠与携带Cre基因的小鼠交配时的重组酶 以转基因为目标。重组，因此过度表达 SF-1和CREB仅见于双侧卵巢支持细胞和颗粒细胞。 转基因小鼠。

Sertoli and granulosa cells form an important somatic component of the gonads. These cells help regulate germ cell development by providing essential nutrients and regulatory signals that support their maturation. In turn, Sertoli and granulosa cells are influenced by a host of cellular associations and intricate sets of endocrine and paracrine signals. The complexities of this system serve to underscore the need to develop animal models to study how Sertoli and granulosa cell factors impact gonadal function. Such studies will ultimately improve approaches for birth control and for treating gonadal abnormalities and infertility. SF-1 and CREB emerge as proteins critical for Sertoli and granulosa cell function. These proteins, like many others, have been characterized within these cells but their role in mediating germ cell development remains obscure. This proposal tests the importance of these proteins, while developing a new transgenic approach that will specifically over- express proteins in Sertoli and granulosa cells. This work will not only provide valuable animal models to elucidate gonadal functions of SF-1 and CREB but will yield important technology that is advantageous to many, as it provides a means to create animal models that express elevated levels of proteins while using a weak but highly specific promoter. Four lines of transgenic mice will be created. Two will contain transgenes in which the Cre recombinase gene is expressed from the follicle-stimulating hormone receptor promoter. This promoter is active only in Sertoli and granulosa cells of the gonads and therefore Cre recombinase will reside only in these cells of transgenic mice. Two other lines f mice will contain targeting transgenes designed to over- express either SF-1 or a dominant negative form of CREB from a strong ubiquitous promoter. The genes for these proteins will not be active unless a recombination event occurs which is facilitated by Cre recombinase when Cre expressing mice are mated with mice harboring the targeting transgenes. Recombination and therefore over-expression of SF-1 and CREB will occur only in Sertoli and granulosa cells of doubly transgenic mice.