Identification of circulatory microRNA based biomarkers for early pregnancy diagnosis in buffalo

IF 4.6 2区 生物学 Q2 CELL BIOLOGY
Kanisht Batra, Anju Sehrawat, Aman Kumar, Man Singh, Ramandeep Kaur, Dipin Chander Yadav, Neha Singh, Sushila Maan
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Abstract

Introduction: The most crucial factor in improving animal reproduction efficiency is early pregnancy diagnosis. Early diagnosis not only reduces the time interval between two calvings but also aids farmers in identifying open animals, thereby preventing significant milk production losses. Therefore, the objective of this study was to discover circulatory miRNAs that would be useful for early pregnancy diagnosis in buffalo.Material and methods: Blood samples were taken on 0, 6th, 12th, and 18th day after artificial insemination from pregnant animals (n = 30) and non-pregnant animals (n = 20). During these stages of pregnancy, total RNA was extracted, and a small RNA library was subsequently generated and sequenced on the Illumina platform. Subsequently, Real-time PCR was used to validate the findings.Results and discussion: There were 4,022 miRNAs found during the pregnancy, with 15 of those lacking sequences and 4,007 having sequences already in the database. From the beginning of pregnancy until the 18th day, 25 of these miRNAs showed a substantial shift in expression levels in the maternal blood, with a change more than two logs. Furthermore, based on qPCR results, 19 miRNAs were found to be more abundant in pregnant animals than in non-pregnant animals. We used target prediction analysis to learn how maternally expressed miRNAs relate to fetal-maternal communication. In conclusion, miRNA based biomarkers that could be associated with the diagnosis of pregnancy were identified including miR-181a and miR-486 highly upregulated on the 18th day of pregnancy. This study also provides a comprehensive profile of the entire miRNA population in maternal buffalo blood during the early stages of pregnancy.
基于循环微RNA生物标记的水牛早孕诊断鉴定
简介提高动物繁殖效率的最关键因素是早期妊娠诊断。早期诊断不仅能缩短两次产犊之间的时间间隔,还能帮助养殖户识别开放性动物,从而避免重大的产奶量损失。因此,本研究旨在发现有助于水牛早期妊娠诊断的循环 miRNA:在水牛人工授精后的第 0 天、第 6 天、第 12 天和第 18 天采集妊娠水牛(30 头)和非妊娠水牛(20 头)的血样。在这些妊娠阶段提取总 RNA,随后生成小 RNA 文库,并在 Illumina 平台上进行测序。随后,使用 Real-time PCR 验证研究结果:孕期共发现 4,022 个 miRNA,其中 15 个缺乏序列,4,007 个在数据库中已有序列。从怀孕开始到第 18 天,其中 25 个 miRNA 在母体血液中的表达水平出现了大幅变化,变化幅度超过两个对数值。此外,根据 qPCR 结果,我们发现有 19 种 miRNA 在妊娠动物体内的表达量高于非妊娠动物。我们利用目标预测分析来了解母体表达的 miRNA 与胎儿和母体交流的关系。总之,研究发现了与妊娠诊断相关的 miRNA 生物标志物,包括在妊娠第 18 天高度上调的 miR-181a 和 miR-486。这项研究还提供了怀孕早期母水牛血液中整个 miRNA 群体的综合概况。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Frontiers in Cell and Developmental Biology
Frontiers in Cell and Developmental Biology Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
9.70
自引率
3.60%
发文量
2531
审稿时长
12 weeks
期刊介绍: Frontiers in Cell and Developmental Biology is a broad-scope, interdisciplinary open-access journal, focusing on the fundamental processes of life, led by Prof Amanda Fisher and supported by a geographically diverse, high-quality editorial board. The journal welcomes submissions on a wide spectrum of cell and developmental biology, covering intracellular and extracellular dynamics, with sections focusing on signaling, adhesion, migration, cell death and survival and membrane trafficking. Additionally, the journal offers sections dedicated to the cutting edge of fundamental and translational research in molecular medicine and stem cell biology. With a collaborative, rigorous and transparent peer-review, the journal produces the highest scientific quality in both fundamental and applied research, and advanced article level metrics measure the real-time impact and influence of each publication.
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