Preconception Paternal Alcohol Exposure Decreases IVF Embryo Survival and Pregnancy Success Rates in a Mouse Model. Preconception Paternal Alcohol Exposure Decreases IVF Embryo Survival and Pregnancy Success Rates in a Mouse Model

In this study, we describe the use of high-throughput sequencing to investigate the developmental basis of preimplantation embryos sired by alcohol-exposed males through in vitro fertilization (IVF). We exposed C57BL/6J male mice to one of three treatment groups. Sexually mature males were randomly selected to voluntarily consume 0% ethanol (control), 6% ethanol, or 10% ethanol for ten weeks. After treatment, the males were sacrificed for sperm collection and cryopreservation for IVF procedures. C57BL/6J donor females remained naïve to alcohol and were synchronized and superovulated before being sacrificed for oocyte collection. Oocytes underwent IVF with the cryopreserved semen and incubated until they reached the morula stage. The morulae sired by ethanol-exposed sperm exhibited altered transcriptional signatures compared to the control-sired morulae. Quick Biology Inc. sequencing core isolated total RNA from three pools (10-15 cells) of unsexed morulae from each control and ethanol preconception treatment. After obtaining the sequencing data, we conducted deep-sequencing analyses of the morulae transcriptomes. This experiment revealed sire alcohol consumption modifies preimplantation embryo genetic pathways controlling mitochondrial dysfunction and oxidative phosphorylation. This experiment furthers our understanding of growth and development changes induced by preconception paternal exposures. Overall design: Examination of paternal preconception alcohol consumption influence on gene expression in IVF morula stage embryos.

本研究采用高通量测序（high-throughput sequencing）技术，探究酒精暴露雄性小鼠通过体外受精（in vitro fertilization, IVF）所产生的植入前胚胎的发育基础。我们将C57BL/6J品系雄性小鼠分为三个处理组：随机选取性成熟雄性个体，使其自由饮用0%乙醇（对照组）、6%乙醇或10%乙醇，持续十周。处理结束后，处死雄性小鼠以获取精子，并将精子冷冻保存以备体外受精实验使用。C57BL/6J品系供体雌性小鼠未接触过酒精，在处死获取卵母细胞前，先对其进行同期化处理与超数排卵。将卵母细胞与冷冻保存的精子进行体外受精，随后培养至桑椹胚阶段。与对照组精子所获得的桑椹胚相比，酒精暴露组精子产生的桑椹胚呈现出显著改变的转录特征。Quick Biology Inc.测序中心分别从对照组与孕前酒精暴露处理组的未分型桑椹胚中，制备3份混合样本（每份样本包含10~15个未分性别的桑椹胚细胞），并从中提取总RNA。获取测序数据后，我们对桑椹胚的转录组开展了深度测序分析。本实验揭示，雄性亲本的酒精摄入会改变植入前胚胎中调控线粒体功能异常与氧化磷酸化的遗传通路。本实验加深了我们对孕前父本暴露所诱导的生长发育改变的认知。实验整体设计：探究孕前父本酒精摄入对体外受精（IVF）桑椹胚阶段胚胎基因表达的影响。