Impact of Egg Quality on Gene Expression and Behavior

描述（由申请人提供）：现在从卵子捐赠研究中清楚地认识到，是卵子而不是生殖道导致生育能力下降或不孕症。值得注意的是，构成“高”品质鸡蛋的分子基础基本上是未知的。Specific Aim 1将通过抑制减法杂交来鉴定在减数分裂和发育正常的高质量小鼠卵子中差异表达的基因，从而验证高质量卵子具有特定基因表达模式的假设。这些基因将用于构建“卵母细胞芯片”，用于确定这些基因的特定集合是否在发育能力受损的卵母细胞中错误表达或不适当表达，例如，在体外培养过程中发育的卵母细胞；卵母细胞来自小的窦卵泡，卵母细胞来自过度刺激的小鼠，卵母细胞来自老年小鼠。成人疾病的胎儿起源假说认为，成人的特定疾病是胎儿在子宫内适应母体营养不足或营养不良的结果。这一假设可以延伸到胚胎着床前阶段，因为在这段时间内减少大鼠的蛋白质摄入量会导致这种长期的变化，而胚胎着床前的培养会导致后代表现出行为改变。具体目标2将通过对这些后代进行一系列行为分析，来验证低质量卵子会产生表现出行为紊乱的后代的假设。用辅助生殖技术（ART）治疗人类不孕症需要胚胎培养，这可能导致基因表达的变化。即使是用于抗逆转录病毒治疗的高质量卵子（以及由此产生的植入前胚胎）也可能仅仅由于胚胎培养条件而处于危险之中。特异性目标3将验证这样一个假设，即改变培养条件可以减轻胚胎培养对后代基因表达和行为中观察到的培养相关扰动的影响。这些实验的结果将建立卵子质量的分子基础，并将提供有关低质量卵子对后代行为的影响的关键信息，以及ART程序对基因表达和行为的影响。

DESCRIPTION (provided by applicant): It is now clearly recognized from egg donation studies that it is the egg and not the reproductive tract that is the cause of reduced fertility or infertility. Remarkably, the molecular underpinnings of what constitutes an egg of "high" quality are essentially unknown. Specific Aim 1 will test the hypothesis that a specific pattern of gene expression characterizes eggs of high quality by using suppression subtractive hybridization to identify genes that are differentially expressed in meiotically and developmentally competent high quality mouse eggs. These genes will be used to construct an "oocyte chip" that will be used to ascertain if a specific set of these genes is mis-expressed, or inappropriately expressed, in oocytes whose developmental competence is compromised, e.g., oocytes that develop during in vitro culture; oocytes derived from small antral follicles, oocytes obtained from hyperstimulated mice, and oocytes obtained from aged mice. The Fetal Origins of Adult Disease hypothesis posits that specific diseases in the adult are the result of the fetus's adaptations in utero to maternal under nutrition or malnutrition. This hypothesis may extend to the preimplantation stage, since reducing protein intake during this time in the rat results in such long-term changes, and the culturing of preimplantation embryos leads to offspring that exhibit altered behavior. Specific Aim 2 will test the hypothesis that low quality eggs give rise to offspring that exhibit perturbations in behavior by analyzing these offspring with a battery of behavioral assays. Treatment of human infertility with Assisted Reproductive Technology (ART) entails embryo culture that can result in changes in gene expression. Even eggs of high quality (and the resulting preimplantation embryos) that are used in ART may be at risk simply due to embryo culture conditions. Specific Aim 3 will test the hypothesis that modifications of culture conditions can alleviate the effect of embryo culture on the culture-associated perturbations observed in gene expression and behavior in the offspring. Results of these experiments will establish the molecular basis of egg quality and will provide critical information regarding the effect on behavior in offspring derived from low quality eggs, as well as the effect of ART procedures on gene expression and behavior.