Lactation and Immuno-Physiology Laboratory, ICAR-National Dairy Research Institute, Karnal, Haryana 132001, India.
Department of Animal Biochemistry, ICAR-National Dairy Research Institute, Karnal, Haryana 132001, India.
J Reprod Immunol. 2020 Aug;140:103148. doi: 10.1016/j.jri.2020.103148. Epub 2020 May 19.
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.
早期且准确地诊断奶牛妊娠对于缩短产犊间隔和提高繁殖效率至关重要。中性粒细胞是最早对子宫内胚胎产生敏感并在干扰素 tau (IFNτ) 诱导下参与母体对妊娠识别的血细胞。为了研究蛋白丰度比值，在人工授精后第 0 天、第 10 天、第 18 天和第 36 天采集杂交卡里弗 Fries 奶牛的血液样本。通过密度梯度离心分离中性粒细胞，并通过高效液相色谱-质谱联用 (LC-MS) 研究其蛋白丰度。结果发现，MX1 和 MX2 两种抗病毒蛋白在授精后第 10 天和第 18 天的丰度更高（P < 0.05），而寡聚腺苷酸合成酶-1 (OAS1) 和干扰素刺激基因 15 泛素样修饰物 (ISG15) 蛋白在授精后第 18 天的丰度更高。还通过 qPCR 研究了这些分子的相对 mRNA 表达。结果表明，与授精后第 0 天相比，ISG15、MX1、MX2 和 OAS1 的基因表达在授精后第 10 天、第 18 天和第 36 天均更高（P < 0.05）。该研究表明，血液中性粒细胞中的 ISGs 对妊娠的建立至关重要，可作为奶牛妊娠诊断的潜在生物标志物。
