Molecular Endocrinology, Functional Genomics & Systems Biology Laboratory, Animal Biochemistry Division; ICAR-National Dairy Research Institute, Karnal, Haryana-132001, India.
Molecular Endocrinology, Functional Genomics & Systems Biology Laboratory, Animal Biochemistry Division; ICAR-National Dairy Research Institute, Karnal, Haryana-132001, India.
Reprod Biol. 2022 Jun;22(2):100641. doi: 10.1016/j.repbio.2022.100641. Epub 2022 May 4.
Successful reproductive management of buffaloes depends primarily upon timely estrus identification. However, 50% of the estrus events are undetected in buffaloes with the available estrus identification methods, leading to huge financial loss to buffalo farmers. Hence, there is an urgent need to develop an alternative and accurate estrus identification method, particularly on the basis of biomarkers in non-invasive fluids. Thus, the present study aimed to identify RNA based estrus biomarkers in cell free saliva in Bubalus bubalis, so that they can be used for future field applicable RT-LAMP colour reactions. RNA-Seq analysis of cell free salivary RNA showed 49 differentially abundant mRNAs between the estrus and diestrus stages. Among five mature miRNAs predicted from the RNA-Seq data, four were found differentially altered at the estrus stage than the diestrus stage. Validation study by direct salivary transcript analysis (DSTA) on 6 selected mRNAs (PPARGC1a, TIMP1, PEBP4, CSPG5, PRHR and ATOH7) and 5 miRNAs (bta-miR-92b, bta-miR-302d, bta-miR-141, bta-miR-27a and bta-let-7a-5p) showed significantly higher levels of TIMP1 (3.46 fold; P < 0.5) and bta-mir-141 (1.33 fold; P < 0.5) in cell-free saliva at the estrus stage compared to the diestrus stage. Hence, TIMP1 and miR-141 appear to be the possible transcript biomarkers for estrus in the cell free saliva of the buffalo. However, further validation studies are required in a large population of buffaloes to determine their estrus biomarker potential before considering them for RT-LAMP colour reaction.
水牛的繁殖管理成功主要取决于及时的发情鉴定。然而，现有的发情鉴定方法有 50%的发情事件无法检测到，这给水牛养殖户带来了巨大的经济损失。因此，迫切需要开发一种替代的、准确的发情鉴定方法，特别是基于非侵入性体液中的生物标志物。因此，本研究旨在鉴定布氏瘤牛无细胞唾液中的基于 RNA 的发情生物标志物，以便将来用于现场适用的 RT-LAMP 颜色反应。无细胞唾液 RNA 的 RNA-Seq 分析显示，发情期和间情期之间有 49 个差异丰富的 mRNA。从 RNA-Seq 数据中预测的五个成熟 miRNA 中，有四个在发情期比间情期有差异改变。通过直接唾液转录分析（DSTA）对 6 个选定的 mRNA（PPARGC1a、TIMP1、PEBP4、CSPG5、PRHR 和 ATOH7）和 5 个 miRNA（bta-miR-92b、bta-miR-302d、bta-miR-141、bta-miR-27a 和 bta-let-7a-5p）进行验证研究表明，TIMP1（3.46 倍；P<0.05）和 bta-miR-141（1.33 倍；P<0.05）在发情期的无细胞唾液中的水平明显高于间情期。因此，TIMP1 和 miR-141 似乎是水牛无细胞唾液中发情的可能转录生物标志物。然而，需要在大量水牛中进行进一步的验证研究，以确定它们作为 RT-LAMP 颜色反应的发情生物标志物的潜力。
