Gecaj Rreze M, Behluli Behlul, Youngs Curtis R
Department of Animal Biotechnology, Faculty of Agriculture and Veterinary, University of Pristina, 10000 Prishtina, Kosovo.
Department of Veterinary Medicine, Faculty of Agriculture and Veterinary, University of Prishtina, 10000 Pristina, Kosovo.
Curr Issues Mol Biol. 2024 Jun 27;46(7):6620-6632. doi: 10.3390/cimb46070394.
In cattle, the corpus luteum (CL) is pivotal in maintaining early pregnancy by secreting progesterone. To establish pregnancy, the conceptus produces interferon-τ, preventing luteolysis and initiating the transformation of the CL spurium into a CL verum. Although this transformation is tightly regulated, limited data are available on the expression of microRNAs (miRNAs) during and after this process. To address this gap, we re-analyzed previously published RNA-Seq data of CL from pregnant cows and regressed CL from non-pregnant cows. This analysis identified 44 differentially expressed miRNAs. From this pool, three miRNAs-bta-miR-222-3p, bta-miR-29c, and bta-miR-2411-3p-were randomly selected for relative quantification. Using bovine ovaries (n = 14) obtained from an abattoir, total RNA (including miRNAs) was extracted and converted to cDNA for RT-qPCR. The results revealed that bta-miR-222-3p was downregulated ( = 0.016) in pregnant females compared to non-pregnant cows with regressed CL. However, no differences in miRNA expression were observed between CL of pregnant and non-pregnant cows for bta-miR-29c ( > 0.32) or bta-miR-2411-3p ( > 0.60). In silico prediction approaches indicated that these miRNAs are involved in pathways regulating pregnancy maintenance, such as the VEGF- and FoxO-signaling pathways. Additionally, their biogenesis is regulated by GABPA and E2F4 transcription factors. The validation of selected miRNA expression in the CL during pregnancy by RT-qPCR provides novel insights that could potentially lead to the identification of biomarkers related to CL physiology and pregnancy outcome.
在牛中，黄体（CL）通过分泌孕酮在维持早期妊娠中起关键作用。为了建立妊娠，孕体产生干扰素-τ，防止黄体溶解并启动假黄体向真黄体的转变。尽管这种转变受到严格调控，但关于在此过程期间和之后微小RNA（miRNA）表达的数据有限。为了填补这一空白，我们重新分析了先前发表的怀孕母牛CL的RNA测序数据以及未怀孕母牛退化CL的RNA测序数据。该分析鉴定出44个差异表达的miRNA。从这个库中，随机选择了3个miRNA——bta-miR-222-3p、bta-miR-29c和bta-miR-2411-3p——进行相对定量。使用从屠宰场获得的牛卵巢（n = 14），提取总RNA（包括miRNA）并转化为用于RT-qPCR的cDNA。结果显示，与退化CL的未怀孕母牛相比，怀孕母牛中bta-miR-222-3p表达下调（P = 0.016）。然而，对于bta-miR-29c（P > 0.32）或bta-miR-2411-3p（P > 0.60），怀孕和未怀孕母牛的CL之间未观察到miRNA表达差异。计算机预测方法表明，这些miRNA参与调节妊娠维持的途径，如VEGF和FoxO信号通路。此外，它们的生物合成受GABPA和E2F4转录因子调控。通过RT-qPCR对怀孕期间CL中选定miRNA表达的验证提供了新的见解，这可能潜在地导致鉴定与CL生理学和妊娠结局相关的生物标志物。
