ANALYSIS OF REPRODUCTIVE FUNCTION USING TRANSGENIC MICE

生殖发育和功能是需要性腺外因素的复杂过程(例如， 促卵泡激素和促黄体生成素)和性腺内因子(如类固醇和多肽生长因子)。女性的生育能力取决于 调节卵母细胞的分配、生长和成熟，这必须与颗粒细胞和膜细胞相协调 卵泡单位内的增殖和分化。在这个过程中，有几个转变 如下：1)未知的静止的原始卵泡向初级(一层)卵泡的募集 机制：2)初级卵泡颗粒细胞生长形成两层和多层 次级卵泡；3)有腔卵泡的形成和细胞的进一步生长，这是一个需要FSH的过程； (4)腔卵泡向排卵前卵泡的过渡，其中有壁状颗粒细胞和卵丘 颗粒细胞具有独特的功能，这是一个由黄体生成素诱导的过程。我们目前资助的竞争续订 (1996-现在)假设转化生长因子p超家族成员，生长分化 因子-9(GDF-9)是卵母细胞第二次和第四次转变所必需的关键的卵子分泌因子。 在3a型初级卵泡期开始的卵母细胞中特异表达，并通过 排卵。利用GDF-9基因敲除小鼠和重组小鼠GDF-9，我们证实了上述假设。 GDF-9基因敲除小鼠由于初级卵泡阶段卵泡发生受阻而不育，并重组 GDF-9可以替代卵母细胞对排卵前期时空表达的基因进行调控。 卵泡。因此，GDF-9是第一个被鉴定的具有多功能的卵母细胞分泌的生长因子。 在调节卵巢体细胞功能和基因表达方面。在这项竞争中描述的研究 更新方案将继续使用这些关键试剂(即GDF-9基因敲除小鼠和重组GDF-9) 并建立更多的转基因小鼠模型和试剂，以进一步确定GDF-9和其他 颗粒细胞和膜细胞生长分化的基本调节剂。

Reproductive development and function are complex processes requiring both extragonadal factors (e.g., FSH and LH) and intragonadal factors (e.g., steroids and peptide growth factors). Female fertility depends on the regulated allocation, growth, and maturation of oocytes, which must be coordinated with granulosa and theca cell proliferation and differentiation within the ovarian follicular unit. During this process, there are several transitions as follows: 1) Recruitment of quiescent primordial follicles to primary (one layer) follicles by an unknown mechanism; 2) Growth of the granulosa cells of the primary follicle to form two-layered and multi-layered secondary follicles; 3) Formation of antral follicles and further growth of the cells, a process which requires FSH; and 4) Transition of the antral follicle to a preovulatory follicle, in which mural granulosa cells and cumulus granulosa cells take on unique functions, a process induced by LH. Our currently funded competitive renewal (1996-present) hypothesized that the transforming growth factor p superfamily member, growth differentiation factor-9 (GDF-9), is a key oocyte-secreted factor required for transitions 2 and 4. GDF-9 mRNA and protein are specifically expressed within the oocyte beginning at the type 3a primary follicle stage and expressed through ovulation. Using GDF-9 knockout mice and recombinant mouse GDF-9, we have confirmed the above hypothesis. GDF-9 knockout mice are infertile due to a block in folliculogenesis at the primary follicle stage, and recombinant GDF-9 can substitute for the oocyte to regulate genes which are spatiotemporally expressed in the preovulatory ovarian follicle. Thus, GDF-9 is the first oocyte-secreted growth factor identified which plays multifunctional roles in the regulation of ovarian somatic cell function and gene expression. The studies described in this competitive renewal proposal will continue to use these key reagents (i.e., the GDF-9 knockout mice and recombinant GDF-9) and generate additional transgenic mouse models and reagents to further define the functions of GDF-9 and other essential regulators in granulosa cell and theca cell growth and differentiation.