SF1 & CREB: GONADAL ROLES BY A NEW TRANSGENIC APPROACH

支持细胞和颗粒细胞是卵巢的重要组成部分。 生殖腺 这些细胞通过提供 必需的营养素和调节信号，支持他们的 成熟 反过来，支持细胞和颗粒细胞受到一种 大量的细胞组织和复杂的内分泌系统， 旁分泌信号 这个系统的复杂性强调了 有必要开发动物模型来研究支持和颗粒细胞 细胞因子影响性腺功能。 这些研究最终将 用于控制生育和治疗性腺疾病改进方法 畸形和不育。 SF-1和CREB成为支持细胞和颗粒细胞的关键蛋白 功能 这些蛋白质，像许多其他蛋白质一样， 但它们在调节生殖细胞发育中的作用 仍然模糊。 这项提案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， 在支持细胞和颗粒细胞中表达蛋白质。 这项工作将不会 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， 对许多人来说，因为它提供了一种创造动物模型的方法， 蛋白质水平升高，同时使用弱但高度特异性的 启动子 将建立四种转基因小鼠品系。 两个将包含 转基因，其中Cre重组酶基因从重组子表达， 促卵泡激素受体启动子。 该启动子具有活性 只有在支持细胞和颗粒细胞的性腺，因此Cre 重组酶将仅存在于转基因小鼠的这些细胞中。 两 其他小鼠品系将含有设计为过度- 表达SF-1或CREB的显性阴性形式， 泛在启动子这些蛋白质的基因不会被激活 除非发生由Cre促进的重组事件 当表达Cre的小鼠与携带重组酶的小鼠交配时， 针对转基因。 因此，过度表达 SF-1和CREB仅在双胚卵巢的支持细胞和颗粒细胞中表达， 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.