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

摘要 父系环境暴露与生殖结局：体外比较 体内受精 问题陈述：900名儿童橙剂脊柱裂被曝光 越南退伍军人获得退伍军人管理局推定服务相关福利。发展终点 如结构性出生缺陷、妊娠失败、低出生体重、儿童癌症增加； 据报道，F1和F2出现了发育、内分泌和生化异常 父亲暴露于杀虫剂、除草剂和辐射后的几代人。多重 与男性介导的不良生殖事件有关的作用机制包括： A)精液中可导致阴道吸收和暴露胚胎的毒素 受孕；b)有害影响，可通过与 神经内分泌轴。据我们所知，没有研究研究父亲的先入为主的作用 暴露于环境因素对精子分离相关生殖结局的影响 通过体外受精和体内受精的比较，得出精液因素的影响因素。 研究目的：主要目的是描述男性先入为主的影响 二恶英暴露对自然交配和体外受精选定参数的影响。小鼠 体外受精绕过了女性精液接触，从而分离了 怀孕后阴道对毒素的吸收和胚胎对这些毒素的暴露 不会对雄性生殖细胞产生任何遗传影响。次要目标是确定芳香族 碳氢受体(AHR)激活对葡萄糖转运体(GLUT)1定位的影响 囊胚期胚胎或子宫环境中的基质细胞，因此具有 分别对胚胎发育和/或植入造成不利影响。AHR激活 已经被证明改变了胚胎细胞系中的GLUT 1的定位，从而导致 并为二恶英的胚胎毒性作用提供了一种解释。 研究方法：试验性2×2区组设计，治疗后T+14和T+35天 将被用来描述体内自然交配产生的生殖结果 未接触二恶英的女性与未接触二恶英的男性的体外受精比较。在……里面 活体生殖测量包括后3.5天(DPC)：a)囊胚数；b) 胚泡TUNEL、细胞色素P1A1和GLUT水平及定位；c)体内胚泡的RT-PCR 和间质细胞CYP1A1基因的表达，以及d)间质细胞蜕膜化。体内和体外 参数包括：a)14.5dpc胎盘和胎儿体重，以及从头到尾的长度；b)全长 足月产仔数、出生体重和5周生长率；c)大体和微观结构。

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.