An early and precise diagnosis of pregnancy in cows is critical to short the calving interval and to improve their reproductive efficiency. Neutrophils are the first blood cells to sensitize the embryo in the uterus and participate in maternal recognition of pregnancy after getting induced by interferon tau (IFNτ). To study the protein abundance ratio, blood samples were collected on 0th, 10th, 18th and 36th day post-artificial insemination (AI) from crossbred Karan Fries cows. Neutrophils were isolated through density gradient centrifugation and studied for protein abundance by high-performance liquid chromatography coupled with mass spectrometry (LC-MS). Protein abundance ratios for Myxovirus resistance (MX1 and MX2) were found to be higher (P < 0.05) on day 10 and day 18 post-AI, whereas Oligoadenylate synthetase-1 (OAS1) and Interferon stimulated gene-15 ubiquitin-like modifier (ISG15) proteins were more abundant on day 18 post-AI. The relative mRNA expressions of these molecules were also studied by qPCR. The gene expression of ISG15, MX1, MX2 and OAS1 was found to be higher (P < 0.05) on day 10th, 18th and 36th post-AI compared to day 0. The study indicates that ISGs on blood neutrophils are essential for the establishment of pregnancy and may be targeted as potential biomarkers for pregnancy diagnosis in cows.

中文翻译：

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蛋白质组学和转录组学研究揭示了 ISG 作为新分子用于奶牛早期妊娠诊断的效用。

奶牛妊娠的早期准确诊断对于缩短产犊间隔和提高繁殖效率至关重要。中性粒细胞是第一个使子宫内的胚胎敏感并在被干扰素 tau (IFNτ) 诱导后参与母体识别妊娠的血细胞。为了研究蛋白质丰度比，在人工授精 (AI) 后第 0、10、18 和 36 天从杂交卡兰薯条奶牛收集血液样本。通过密度梯度离心分离中性粒细胞，并通过高效液相色谱与质谱联用 (LC-MS) 研究蛋白质丰度。发现粘病毒抗性（MX1 和 MX2）的蛋白质丰度比在 AI 后第 10 天和第 18 天更高（P < 0.05），而寡腺苷酸合成酶-1 (OAS1) 和干扰素刺激的基因 15 泛素样修饰剂 (ISG15) 蛋白在 AI 后第 18 天更为丰富。还通过 qPCR 研究了这些分子的相对 mRNA 表达。与第 0 天相比，在 AI 后第 10、18 和 36 天发现 ISG15、MX1、MX2 和 OAS1 的基因表达更高（P < 0.05）。该研究表明，血液中性粒细胞上的 ISG 对于建立妊娠，并可能成为奶牛妊娠诊断的潜在生物标志物。