Prairie View A&M University, Cooperative Agricultural Research Center, Prairie View, TX, 77446-0519, USA.
Prairie View A&M University, Cooperative Agricultural Research Center, Prairie View, TX, 77446-0519, USA.
Theriogenology. 2019 Jul 1;132:118-127. doi: 10.1016/j.theriogenology.2019.04.022. Epub 2019 Apr 16.
Regulation of the expression of the alpha(1,2)fucosyltransferase (FUT) genes and their enzymatic products, including the H-type 1 antigen (HT1), on the luminal surface of the uterus is believed to be critical for establishment of pregnancy in mammals. The FUT1 gene is a marker for conception rates in dairy cows and HT1 is a marker for uterine receptivity in rodents. To determine the spatiotemporal expression patterns of FUT1 and FUT2 genes in goats, endometrial tissues were obtained on six days spanning the estrous cycle (Days 5, 11, 13, 15, 17 and 19) and seven days spanning early pregnancy (Days 5, 11, 13, 15, 17, 19 and 25). In all data, we found no effect of status (cyclic or pregnant; P > 0.1) and pooled data where appropriate. We cloned FUT1 cDNA from goat endometrium and made probes from it for Northern and slot blot analyses. The analyses indicated that FUT1 gene expression was high until Day 13, and then declined. In situ hybridization revealed a change in the cell-specificity of FUT1 gene expression over the estrous cycle and early pregnancy. In situ hybridization signal intensity scores indicated that FUT1 expression by uterine epithelium was high on Day 5, moderate on Day 11, and minimal on subsequent days. In situ hybridization signals in uterine glandular epithelial cells remained high from Day 5 to Day 13, with weaker signals thereafter. Quantitative reverse transcription-PCR (RT-qPCR) assays were used for quantitation of FUT1 and FUT2 mRNAs. Quantitative RT-qPCR data were generated from endometrium collected from cyclic and pregnant animals on Days 5, 11 and 17. Relative levels of FUT1 mRNA were high on Days 5 and 11, but then fell 5-fold by Day 17 (P < 0.01). FUT2 mRNA concentrations were below the accurate detectable limit of the assay. High levels of HT1 were observed on the apical surface of uterine luminal epithelia on Days 5, 15, 17 and 19, with much lower levels on Days 11 and 13. Thus, data suggests that FUT1 is the primary enzyme responsible for the high levels of HT1 antigen present on the uterine luminal epithelium between Days 5 and 11 of the estrous cycle and early pregnancy. But changes in the expression of the FUT1 gene does not directly correlate to HT1 staining, which increased from Day 13-15. Future studies are required to understand the regulation of the HT1 antigen on the luminal surface of endometrium.
调控子宫腔表面的α(1,2)岩藻糖基转移酶（FUT）基因及其酶产物的表达，包括 H 型 1 抗原（HT1），被认为对哺乳动物妊娠的建立至关重要。FUT1 基因是奶牛受孕率的标志物，HT1 是啮齿动物子宫容受性的标志物。为了确定 FUT1 和 FUT2 基因在山羊中的时空表达模式，在发情周期（第 5、11、13、15、17 和 19 天）和早孕（第 5、11、13、15、17、19 和 25 天）期间获得子宫内膜组织。在所有数据中，我们都没有发现状态（循环或怀孕；P > 0.1）的影响，并在适当的情况下汇总数据。我们从山羊子宫内膜中克隆了 FUT1 cDNA，并从它制作了 Northern 和 slot blot 分析探针。分析表明，FUT1 基因表达直到第 13 天一直很高，然后下降。原位杂交揭示了发情周期和早孕期间 FUT1 基因表达的细胞特异性变化。原位杂交信号强度评分表明，子宫上皮的 FUT1 表达在第 5 天很高，第 11 天适中，随后几天则很低。子宫腺上皮细胞中的原位杂交信号从第 5 天到第 13 天保持较高水平，随后信号减弱。定量逆转录聚合酶链反应（RT-qPCR）检测用于定量 FUT1 和 FUT2 mRNA。使用从发情周期和怀孕动物在第 5、11 和 17 天收集的子宫内膜进行定量 RT-qPCR 检测。FUT1 mRNA 的相对水平在第 5 天和第 11 天较高，但到第 17 天下降 5 倍（P < 0.01）。FUT2 mRNA 浓度低于检测限。在发情周期和早孕期间的第 5、15、17 和 19 天，子宫腔上皮的顶端表面观察到高水平的 HT1，而在第 11 和 13 天则水平较低。因此，数据表明 FUT1 是发情周期和早孕期间第 5 至 11 天期间子宫腔上皮表面高 HT1 抗原的主要酶。但是 FUT1 基因表达的变化与 HT1 染色并不直接相关，HT1 染色从第 13-15 天开始增加。需要进一步研究来了解 HT1 抗原在子宫内膜腔表面的调控。
