Maternal tract distinguish between spermatozoa and other pathogenic and non-pathogenic entities. Sus scrofa

The mammalian oviduct is a dynamic organ and the venue of important events leading to the establishment of pregnancy. Oviductal epithelial cells are involved in direct contact and specific interactions with different self and non-self entities (oocyte, sperm, etc.).The aim of this study was to determine if the oviduct is able to distinguish between different types of non-self-entities. We used an in vitro model to determine genes altered in oviductal epithelial cell (OPEC) in response to the presence of spermatozoa, mammalian somatic cells (red blood cell) or bacteria. We hypothesised that alteration of OPEC expression profile in response to boar spermatozoa (friendly non-self entity), red blood cells (non-pathogenic, non-self entity) and bacteria (pathogenic non-self entity) is indicative of recognition of self and non-self entities by these cells as well as recognition of pathogenic from non-pathogenic non-self entities. Overall design: Oviductal epithelial cells (OPEC) were co-incubated in the presence of (i) bacteria (1 x 10˄8 bacteria), (ii) sperm (1 X 10 ˄ 6 spermatozoa/ml) or (iii) red blood cell (5 x 10 ˄ 6 red blood cell/ml) for 24 hours in HAM-12 media. Three biological replicates were performed for each group and a total of 12 Affymetrix Porcine arrays were used to identify the expression profiles of OPEC alone (3 arrays) or in the presence of (i) bacteria (3 arrays), (ii) sperm (3 arrays) or (iii) red blood cells (3 arrays).

哺乳动物输卵管是一类动态器官，亦是妊娠建立相关关键生物学事件的发生场所。输卵管上皮细胞可与多种自体及非自体实体（卵母细胞、精子等）发生直接接触与特异性相互作用。

本研究旨在探究输卵管是否能够区分不同类型的非自体实体。我们采用体外模型，检测输卵管上皮细胞（Oviductal Epithelial Cell, OPEC）在接触精子、哺乳动物体细胞（红细胞）或细菌时的差异表达基因。我们提出如下假说：OPEC在响应公猪精子（友好型非自体实体）、红细胞（非致病性非自体实体）与细菌（致病性非自体实体）时的表达谱改变，可反映该细胞对自体与非自体实体的识别能力，以及对致病性与非致病性非自体实体的区分能力。

实验设计：将OPEC分别与以下物质在HAM-12培养基中共培养24小时：(i) 细菌（1×10^8个）、(ii) 精子（1×10^6个/ml）、(iii) 红细胞（5×10^6个/ml）。每组设置3次生物学重复，共使用12张Affymetrix猪基因芯片（Affymetrix Porcine Arrays），分别检测单独培养的OPEC（3张芯片），以及分别与细菌（3张）、精子（3张）、红细胞（3张）共培养的OPEC的基因表达谱。