A transcriptomal analysis of bovine oviductal epithelial cells collected during the follicular phase versus the luteal phase of the estrous cycle.
A transcriptomal analysis of bovine oviductal epithelial cells collected during the follicular phase versus the luteal phase of the estrous cycle.
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DOI：
10.1186/s12958-015-0077-1
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发表时间：
2015-08-05
期刊：
影响因子：
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通讯作者：
Bridges PJ
中科院分区：
文献类型：
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作者：
Cerny KL;Garrett E;Walton AJ;Anderson LH;Bridges PJ
生殖成功取决于配子储存、成熟、受精和早期胚胎发育的功能性输卵管。卵巢源性类固醇雌激素和黄体酮是输卵管功能的关键调节剂。本研究的目的是通过使用基于微阵列的转录分析来研究黄体和卵泡期特异性输卵管上皮细胞功能，以增加我们对调节上皮细胞过程的 mRNA 的理解，并确定可能在未来影响生育力的新基因和生化途径。六头正常周期的安格斯小母牛被分配到黄体期（LP，n = 3）或卵泡期（FP，n = 3）治疗组。 LP 组的小母牛在发情后第 11 天到第 12 天之间被宰杀。 FP 组的小母牛在发情后第 6 天晚上 8 点接受 25 mg PGF2α（Lutalyse，辉瑞，纽约）治疗，并在 36 小时后处死。使用从 LP 和 FP 治疗组收集的壶腹和峡部切片分离的上皮细胞的总 RNA，通过微阵列进行转录分析，并通过实时 RT-PCR 分析确认选定的 mRNA。对差异表达基因进行基因本体分类和生物信息学通路分析。使用 Benjamini-hochberg 多重检验校正错误发现率 (FDR) 的统计单向方差分析以及 FP 组与 LP 组壶腹部上皮细胞的成对比较显示，分别有 972 个和 597 个转录本上调和下调 (P<0.05)。在 FP 组与 LP 组的峡部上皮细胞内，分别有 946 个和 817 个转录本上调和下调（P<0.05）。壶腹和峡部的上调基因主要参与胆固醇生物合成和细胞周期途径，而下调基因则存在于许多炎症反应途径中。基于微阵列的转录分析揭示了输卵管壶腹和峡部上皮内 mRNA 表达的周期依赖性变化阶段。本文的在线版本 (doi:10.1186/s12958-015-0077-1) 包含补充材料，可供授权用户使用。
Reproductive success depends on a functional oviduct for gamete storage, maturation, fertilization, and early embryonic development. The ovarian-derived steroids estrogen and progesterone are key regulators of oviductal function. The objective of this study was to investigate luteal and follicular phase-specific oviductal epithelial cell function by using microarray-based transcriptional profiling, to increase our understanding of mRNAs regulating epithelial cell processes, and to identify novel genes and biochemical pathways that may be found to affect fertility in the future. Six normally cycling Angus heifers were assigned to either luteal phase (LP, n = 3) or follicular phase (FP, n = 3) treatment groups. Heifers in the LP group were killed between day 11 and 12 after estrus. Heifers in the FP group were treated with 25 mg PGF2α (Lutalyse, Pfizer, NY) at 8 pm on day 6 after estrus and killed 36 h later. Transcriptional profiling by microarray and confirmation of selected mRNAs by real-time RT-PCR analyses was performed using total RNA from epithelial cells isolated from sections of the ampulla and isthmus collected from LP and FP treatment groups. Differentially expressed genes were subjected to gene ontology classification and bioinformatic pathway analyses. Statistical one-way ANOVA using Benjamini-hochberg multiple testing correction for false discovery rate (FDR) and pairwise comparison of epithelial cells in the ampulla of FP versus LP groups revealed 972 and 597 transcripts up- and down-regulated, respectively (P < 0.05). Within epithelial cells of the isthmus in FP versus LP groups, 946 and 817 transcripts were up- and down-regulated, respectively (P < 0.05). Up-regulated genes from both ampulla and isthmus were found to be largely involved in cholesterol biosynthesis and cell cycle pathways, while down-regulated genes were found in numerous inflammatory response pathways. Microarray-based transcriptional profiling revealed phase of the cycle-dependent changes in the expression of mRNA within the epithelium of the oviducts’ ampulla and isthmus. The online version of this article (doi:10.1186/s12958-015-0077-1) contains supplementary material, which is available to authorized users.