Functional genomics of follicular growth and ovulation

本研究旨在鉴定和研究与卵泡选择/优势和排卵有关的控制卵泡和卵母细胞发育的分子决定因素。奶牛被用作工作模型，因为它模拟了在妇女身上观察到的卵泡发育的生理模式，并代表了具有重要经济意义的活体模型。通过应用cDNA消减杂交鉴定的分子决定簇，通过比较未选择的卵泡和优势卵泡，以及优势卵泡和排卵卵泡，可以建立不同卵泡阶段颗粒细胞的基因指数。已知和新发现的基因在卵泡发育和排卵的不同阶段的表达模式将通过RT-PCR进行研究。鉴定出的新的cDNA将通过克隆和测序其全长cDNA来表征。研究与卵泡生长和排卵有关的基因的时空表达模式将增加我们对这一生理过程的理解和控制：识别参与细胞增殖和细胞信号传递(细胞外和细胞内)的新基因，影响组织重塑、愈合和炎症过程；增加用于体外受精、核移植和转基因的合格卵母细胞的产量；提高针对具有重要经济意义的驯养动物的卵泡同步和卵巢超刺激计划以及人类生育和避孕计划的治疗效果；以及确定与育性相关的候选基因，这些候选基因可以作为分子标记在涉及商业化和分配雄配子和雌配子的行业的育种计划中用于遗传辅助选择。

This research program aims at the identification and the study of molecular determinants that control the development of the ovarian follicle and oocyte, in relation to follicular selection/dominance and ovulation. The cow is used as the working model since it mimics the physiological pattern of follicular development observed in women, and represents an in vivo model of economic importance. Molecular determinants identified through the application of cDNA subtractive hybridization has allows the establishment of a gene index for granulosa cells at different follicular stages, by comparing non-selected to dominant follicles, and dominant to ovulatory follicles. The expression pattern of genes (known and novel cDNAs) identified, will be investigated by RT-PCR at different stages of follicular development and ovulation. The novel cDNAs identified, will be characterized by cloning and sequencing of their full-length cDNA. The study of the spatio-temporal expression pattern of genes in relation to follicular growth and ovulation will increase our understanding and control of this physiological process: to identify novel genes involved in cellular proliferation and cell signaling (extra and intra cellular) that impact on tissue remodeling, healing, and inflammatory processes; to increase the production of competent oocytes used for in vitro fertilization, nuclear transfer and transgenesis; to increase the efficacy of treatments aiming at follicle synchronization and ovarian super-stimulation programs in domesticated animals of economic importance, and in fertility and contraception programs in humans; and to identify candidate genes associated with fertility that can be used as molecular markers for genetic assisted selection in breeding programs of industries involved in commercializing and distributing male and female gametes.