Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
Theriogenology. 2012 Feb;77(3):539-49. doi: 10.1016/j.theriogenology.2011.08.029. Epub 2011 Oct 19.
Oviductins belong to a family of oviduct-specific glycoproteins believed to play an important role in fertilization and/or early embryonic development. Oviductin cDNA between species is highly conserved and shares 58% to 98% similarity in the deduced amino acid sequences. Our objective in this study was to sequence the full open reading frame of the feline oviductin and to examine its expression during the estrous cycle on both mRNA and protein level. The obtained cDNA containing the full open reading frame was determined to be 1677 nucleotides coding for a deduced protein of 558 amino acids. Identities between species range from 74% (mouse) to 80% (human, baboon, and rhesus) within the N-terminal protein region. Major differences were localized in the carboxy terminal region, which corresponds to exon 11 of the gene. Feline oviductin contained one putative N-linked glycosylation site, six O-linked glycosylation sites, a potential heparin binding site, and two cholesterol recognition and/or interaction amino acid consensus (CRAC) domains. Oviductin expression was analyzed by real time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry. Both approaches revealed an estrous cycle-dependent expression in the ampulla and isthmus. Quantitative PCR showed highest oviductin mRNA copy numbers in the early and late follicular stage and reduced mRNA expression during all other stages. With the exception of the early follicular stage, feline oviductin mRNA abundance was not significantly different in the oviductal segments ampulla and isthmus. A prominent immunolabeling was seen in the early and late follicular stage which disappeared after ovulation, indicating a function of the protein during sperm storage and fertilization.
卵管蛋白属于输卵管特异性糖蛋白家族,被认为在受精和/或早期胚胎发育中发挥重要作用。种间卵管蛋白 cDNA 高度保守,推导的氨基酸序列具有 58%至 98%的相似性。本研究的目的是对猫的卵管蛋白进行全长开放阅读框测序,并在 mRNA 和蛋白质水平上研究其在发情周期中的表达。获得的 cDNA 含有全长开放阅读框,编码 558 个氨基酸的推导蛋白。种间同一性在 N 端蛋白区域内为 74%(鼠)至 80%(人、狒狒和恒河猴)。主要差异定位于羧基末端区域,对应于基因的第 11 个外显子。猫的卵管蛋白含有一个潜在的 N-连接糖基化位点、六个 O-连接糖基化位点、一个潜在的肝素结合位点和两个胆固醇识别和/或相互作用氨基酸共识(CRAC)结构域。通过实时定量聚合酶链反应(RT-qPCR)和免疫组织化学分析了卵管蛋白的表达。这两种方法都显示出在壶腹和峡部的发情周期依赖性表达。定量 PCR 显示在早卵泡期和晚卵泡期输卵管蛋白 mRNA 拷贝数最高,而在所有其他阶段则降低。除了早卵泡期外,输卵管蛋白 mRNA 丰度在壶腹和峡部的输卵管段之间没有显著差异。在早卵泡期和晚卵泡期观察到明显的免疫标记,排卵后消失,表明该蛋白在精子储存和受精过程中具有功能。