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奶牛在热中性和热应激条件下输卵管上皮细胞转录组和细胞外囊泡的特征。

Oviductal epithelial cells transcriptome and extracellular vesicles characterization during thermoneutral and heat stress conditions in dairy cows.

机构信息

Department of Obstetrics and Reproduction, Veterinary Faculty, University of Thessaly, Karditsa, Greece.

Department of Animal Sciences, University of Thessaly, Larissa, Greece.

出版信息

Theriogenology. 2022 Jul 15;187:152-163. doi: 10.1016/j.theriogenology.2022.04.011. Epub 2022 May 7.

Abstract

In this study, the transcriptome of oviductal epithelial cells and certain characteristics of their extracellular vesicles of dairy cows were described under thermoneutral and heat stress conditions. Twenty cows were compared in springtime at THI = 65.6 ± 0.90 and in summertime at THI = 78.36 ± 2.73. During each season, the estrous cycles of the cows were synchronized, and on day 3 of the ensuing cycle, a blood sample was collected for progesterone determination, while their oviducts were collected after slaughter. Epithelial cells and oviductal fluid were collected from the oviduct ipsilateral and contralateral to the corpus, respectively. For the gene expression study, a comparative transcriptomic approach, using RNASeq, was performed on cells collected from the ipsilateral and the contralateral oviducts. The size and the concentration of extracellular vesicles (EVs) at both seasons were analyzed using Transmission Electron Microscopy and Nanoparticle tracking analysis and specific proteins were detected by Western blotting. Progesterone concentration was higher during the thermoneutral period. Between seasons, divergent expression of genes related to immune system, contractility, gamete protection and lncRNAs was found. The size and the concentration of the EVs did not differ between seasons, however, the concentration in the ipsilateral oviduct tended to be lower (p = 0.09) from the contralateral one in the summer, but not in the spring. Our results show for the first time that HS could be involved with alterations in the oviductal cells' gene expression and in the changes in concentration of EVs in the oviductal lumen. Our results imply that the altered oviductal environment during HS could be associated with the suppressed summer fertility in dairy cows.

摘要

本研究描述了在常温(THI=65.6±0.90)和热应激(THI=78.36±2.73)条件下奶牛输卵管上皮细胞的转录组及其细胞外囊泡的某些特性。在春季和夏季,对 20 头奶牛进行了比较。在每个季节,奶牛的发情周期都被同步化,在随后的周期的第 3 天,采集血液样本以测定孕酮,同时在屠宰后采集其输卵管。从输卵管的同侧和对侧分别采集上皮细胞和输卵管液。为了进行基因表达研究,使用 RNASeq 对同侧和对侧输卵管采集的细胞进行了比较转录组分析。在两个季节,均使用透射电子显微镜和纳米颗粒跟踪分析来分析细胞外囊泡(EVs)的大小和浓度,并通过 Western blotting 检测特定蛋白质。在常温期孕酮浓度较高。在季节之间,与免疫系统、收缩性、配子保护和 lncRNA 相关的基因表达存在差异。EVs 的大小和浓度在两个季节之间没有差异,但是在夏季,同侧输卵管的浓度倾向于低于对侧(p=0.09),而在春季则没有。我们的研究结果首次表明,HS 可能与输卵管细胞基因表达的改变和输卵管腔中 EVs 浓度的变化有关。我们的研究结果表明,HS 期间改变的输卵管环境可能与奶牛夏季生育力下降有关。

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