Paternal Environmental Exposures and Reproductive Outcomes: A Comparison of in Vi

摘要 标题：父亲环境暴露和生殖结局：体外 和体内受精 问题陈述：900名脊柱裂儿童暴露于橙子剂 越南退伍军人获得VA推定服务相关福利。发育终点 如结构性出生缺陷、妊娠失败、出生体重低、儿童癌症增加; 在F1和F2中报告了发育、内分泌和生化异常 父亲暴露于杀虫剂、除草剂和辐射后的后代。多 作用机制涉及男性介导的不良生殖事件，包括： a）精液中的毒素，可导致阴道吸收和胚胎暴露后 B）通过与生殖系统的相互作用而破坏生育能力的有害影响。 神经内分泌轴据我们所知，没有研究探讨父亲的偏见的作用， 暴露于环境因素对生殖结局的影响 通过体外受精和体内受精的比较，从精浆因素中提取影响因素。 研究目的：主要目的是描述男性孕前的影响 二恶英暴露对自然交配和体外受精的选定参数的影响。鼠 体外受精避免了女性精液接触，从而分离了 阴道吸收毒素和胚胎在受孕后暴露于这些毒素 对男性生殖细胞的遗传影响第二个目标是确定芳基是否 碳氢化合物受体（AHR）激活影响葡萄糖转运蛋白（GLUT）1在 胚泡期胚胎或子宫环境的基质细胞，因此具有 分别对胚胎发育和/或植入产生有害影响。AHR活化 已经显示改变胚胎细胞系中的GLUT 1定位，从而导致 营养缺乏的细胞，并提供一个解释二恶英的胚胎毒性作用。 研究方法：在治疗后T+14和T+35天进行实验性2 × 2区组设计 将用于表征体内自然交配产生的生殖结局 vs.未接触二恶英的雌性与接触二恶英和未接触二恶英的雄性进行体外受精。在 体内生殖测量包括术后3.5天（dpc）：a）胚泡数; B） 囊胚TUNEL、CYP 1A 1和GLUT水平和定位; c）体内囊胚的RT-PCR 和基质细胞CYP 1A 1 mRNA，和d）基质细胞蜕膜化。体内和体外 参数包括：a）14.5 dpc胎盘和胎儿重量，以及头臀长度; B）全 足月同窝仔数、出生体重和5周生长率;以及c）大体和显微结构。

Abstract Title: Paternal Environmental Exposures and Reproductive Outcomes: A Comparison of in vitro and in vivo Fertilization Statement of the Problem: Nine hundred children with Spina Bifida of Agent Orange exposed Vietnam veterans receive VA presumptive service connected benefits. Developmental endpoints such as structural birth defects, failed pregnancy, low birth weight, increased childhood cancers; and developmental, endocrine, and biochemical abnormalities have been reported in F1 and F2 generations following paternal exposures to pesticides, herbicides, and radiation. Multiple mechanisms of action are implicated in male mediated adverse reproductive events to include: a) toxins in semen that can result in vaginal absorption and in exposure of the embryo post conception, and b) deleterious effects that can disrupt fertility through interaction with the neuroendocrine axis. To our knowledge no studies examine the role of paternal preconception exposure to environmental agents on the reproductive outcomes by separating sperm related factors from seminal fluid factors through a comparison of in vitro and in vivo fertilization. Research Objectives: The primary objective is to characterize the effects of male preconception dioxin exposure on selected parameters of natural mating as well as in vitro fertilization. Murine in vitro fertilization circumvents female seminal fluid contact thereby separating the effect of vaginal absorption of toxins and the exposure of the embryo to these toxins post conception from any genetic effects on the male germ cells. The secondary objective is to determine if aryl hydrocarbon receptor (AHR) activation affects glucose transporter (GLUT) 1 localization in either the blastocyst stage embryo or stromal cell of the uterine environment, thus having a detrimental effect on embryo development and/or implantation respectively. AHR activation has been shown to alter GLUT 1 localization in an embryonic cell line thereby resulting in nutrient deficient cells and providing one explanation of Dioxin's embryo toxic effects. Research Methods: An experimental 2 x 2 block design at T+14 and T+35 days post treatment will be used to characterize the reproductive outcomes that result from in vivo natural mating vs. in vitro fertilization of non exposed females with Dioxin exposed and non exposed males. In vivo reproductive measures include 3.5 day post coitum (dpc): a) blastocyst numbers; b) blastocyst TUNEL, CYP1A1, and GLUT levels and localization; c) RT-PCR of in vivo blastocyst and stromal cell CYP1A1 mRNA, and d) stromal cell decidualization. In vivo and in vitro parameters include: a) 14.5 dpc placental and fetal weights, and crown to rump lengths; b) full term litter size, birth weights, and 5 week growth rates; and c) gross and microscopic structure.