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微小RNA测序揭示不同水平的非纤维碳水化合物/中性洗涤纤维对犊牛瘤胃发育的影响。

MicroRNA Sequencing Reveals the Effect of Different Levels of Non-Fibrous Carbohydrate/Neutral Detergent Fiber on Rumen Development in Calves.

作者信息

Xue Mingming, Wang Kejun, Wang Ansi, Li Ruiting, Wang Yadong, Sun Shuaijie, Yan Duo, Song Guohua, Xu Huifen, Sun Guirong, Li Ming

机构信息

College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.

出版信息

Animals (Basel). 2019 Jul 28;9(8):496. doi: 10.3390/ani9080496.

DOI:10.3390/ani9080496
PMID:31357699
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6720277/
Abstract

Rumen development in calves is affected by many factors, including dietary composition. MicroRNAs (miRNAs) are known to function in the development of the rumen in cattle, what is not known is how these miRNAs function in rumen development of calves fed with high and low ratios of non-fibrous carbohydrate (NFC)/neutral detergent fiber (NDF). A total of six healthy Charolais hybrids bull calves of similar weight were divided into two groups; three calves were fed a mixed diet with NFC/NDF = 1.35 (H group), and three were fed a mixed diet with NFC/NDF = 0.80 (L group). After 105 days on the diet, calves were sacrificed and rumen tissues were collected. Tissues were subjected to histological observation and miRNA expression analysis. Functional enrichment analysis was conducted on the target genes of the miRNAs. Targeting and regulatory relationships were verified by luciferase reporter assay and quantitative PCR (qPCR). We found that the length of rumen papilla in the L group was significantly greater than that in the H group, while the width of rumen papilla in H group was significantly greater than that that in L group. We identified 896 miRNAs; 540 known miRNAs, and 356 novel predicted miRNAs. After statistical testing, we identified 24 differentially expressed miRNAs (DEmiRNAs). miRNA-mRNA-cluster network analysis and literature reviews revealed that cell proliferation, differentiation, physical and nutrient stimuli processes participate in rumen development under different NFC/NDF levels. The regulatory relationships between three DEmiRNAs and five target genes were verified by examining the levels of expression. The binding sites on bta-miR-128 for the peroxisome proliferator activated receptor gamma () and solute carrier family 16 member 1 ( genes were investigated using a dual luciferase assay. The results of this study provide insight into the role of miRNAs in rumen development in calves under different NFC/NDF levels.

摘要

犊牛的瘤胃发育受多种因素影响，包括日粮组成。已知微小RNA（miRNA）在牛的瘤胃发育中发挥作用，但尚不清楚这些miRNA在不同非纤维碳水化合物（NFC）/中性洗涤纤维（NDF）比例的日粮喂养的犊牛瘤胃发育中如何发挥作用。选取6头体重相近的健康夏洛莱杂交公牛犊，分为两组；3头犊牛饲喂NFC/NDF = 1.35的混合日粮（H组），另外3头饲喂NFC/NDF = 0.80的混合日粮（L组）。日粮饲喂105天后，宰杀犊牛并采集瘤胃组织。对组织进行组织学观察和miRNA表达分析。对miRNA的靶基因进行功能富集分析。通过荧光素酶报告基因检测和定量PCR（qPCR）验证靶向和调控关系。我们发现，L组瘤胃乳头长度显著大于H组，而H组瘤胃乳头宽度显著大于L组。我们鉴定出896个miRNA；其中540个为已知miRNA，356个为新预测的miRNA。经统计学检验，我们鉴定出24个差异表达的miRNA（DEmiRNA）。miRNA-mRNA-聚类网络分析和文献综述表明，细胞增殖、分化、物理和营养刺激过程参与不同NFC/NDF水平下的瘤胃发育。通过检测表达水平验证了3个DEmiRNA与5个靶基因之间的调控关系。利用双荧光素酶检测法研究了过氧化物酶体增殖物激活受体γ（）和溶质载体家族16成员1（基因在bta-miR-128上的结合位点。本研究结果为深入了解miRNA在不同NFC/NDF水平下犊牛瘤胃发育中的作用提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/6bc50fba0d66/animals-09-00496-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/3e2a974f9229/animals-09-00496-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/9690af680b3d/animals-09-00496-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/1da62f8b92f5/animals-09-00496-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/cb140fa6e96c/animals-09-00496-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/0c6473719fba/animals-09-00496-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/476edf028d6f/animals-09-00496-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/6bc50fba0d66/animals-09-00496-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/3e2a974f9229/animals-09-00496-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/9690af680b3d/animals-09-00496-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/1da62f8b92f5/animals-09-00496-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/cb140fa6e96c/animals-09-00496-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/0c6473719fba/animals-09-00496-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/476edf028d6f/animals-09-00496-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/6720277/6bc50fba0d66/animals-09-00496-g007.jpg

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