Exploration of microRNAs in porcine milk exosomes.
Exploration of microRNAs in porcine milk exosomes.
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猪乳外泌体中 microRNA 的探索
DOI：
10.1186/1471-2164-15-100
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发表时间：
2014-02-05
期刊：
影响因子：
4.4
通讯作者：
Zhang YL
中科院分区：
文献类型：
--
作者：
Chen T;Xi QY;Ye RS;Cheng X;Qi QE;Wang SB;Shu G;Wang LN;Zhu XT;Jiang QY;Zhang YL
母乳含有复杂的营养物质，促进婴儿各种生物系统的成熟。外泌体是存在于不同体液（如牛奶）中的内吞起源的膜状囊泡，可介导细胞间通讯。我们假设microRNAs （miRNAs）可能在仔猪的发育过程中发挥重要作用。microRNAs是一类18 - 25nt的非编码小rna，已知存在于人乳、牛乳和猪乳的外泌体中。结果通过连续离心和超离心，从猪乳中分离到直径约为100 nm的外泌体。从外泌体中提取总RNA，发现5S核糖体RNA是主要RNA成分。Solexa测序共发现491个mirna，其中176个为已知mirna， 315个为新的成熟mirna（代表366个pre- mirna），分布在30个簇和35个家族中，并通过荧光素酶测定验证了两个预测的新mirna靶向IGF-1R的3'UTR。有趣的是，我们观察到三种mirna （ssc-let-7e， ssc-miR-27a和ssc-miR-30a）可以由miRNA-offset rna （morna）产生。通过RNAhybrid软件预测，前10个miRNAs占总数的74.5%（67154个），可靶向2333个基因。利用DAVID生物信息学资源对Gene Ontology和Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway进行分析，发现鉴定的mirna靶向转录、免疫和代谢过程富集的基因，前20名mirna中有14个可能参与了IgA免疫网络的调控。结论猪乳外泌体中含有大量mirna，可能在母猪乳汁向仔猪传递信息中发挥重要作用。本研究预测的猪乳外泌体mirna可为今后的生化和生物物理功能研究提供基础。
BackgroundBreast milk contains complex nutrients and facilitates the maturation of various biological systems in infants. Exosomes, membranous vesicles of endocytic origin found in different body fluids such as milk, can mediate intercellular communication. We hypothesized that microRNAs (miRNAs), a class of non-coding small RNAs of 18–25 nt which are known to be packaged in exosomes of human, bovine and porcine milk, may play important roles in the development of piglets.ResultsIn this study, exosomes of approximately 100 nm in diameter were isolated from porcine milk through serial centrifugation and ultracentrifugation procedures. Total RNA was extracted from exosomes, and 5S ribosomal RNA was found to be the major RNA component. Solexa sequencing showed a total of 491 miRNAs, including 176 known miRNAs and 315 novel mature miRNAs (representing 366 pre-miRNAs), which were distributed among 30 clusters and 35 families, and two predicted novel miRNAs were verified targeting 3’UTR of IGF-1R by luciferase assay. Interestingly, we observed that three miRNAs (ssc-let-7e, ssc-miR-27a, and ssc-miR-30a) could be generated from miRNA-offset RNAs (moRNAs). The top 10 miRNAs accounted for 74.5% (67,154 counts) of total counts, which were predicted to target 2,333 genes by RNAhybrid software. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses using DAVID bioinformatics resources indicated that the identified miRNAs targeted genes enriched in transcription, immunity and metabolism processes, and 14 of the top 20 miRNAs possibly participate in regulation of the IgA immune network.ConclusionsOur findings suggest that porcine milk exosomes contain a large number of miRNAs, which potentially play an important role in information transfer from sow milk to piglets. The predicted miRNAs of porcine milk exosomes in this study provide a basis for future biochemical and biophysical function studies.
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DOI：
10.1084/jem.20010670
发表时间：
2002-01-21
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Bowman EP;Kuklin NA;Youngman KR;Lazarus NH;Kunkel EJ;Pan J;Greenberg HB;Butcher EC
通讯作者：
Butcher EC
影响因子：
5.4
作者：
DURANDY, A;SCHIFF, C;FISCHER, A
通讯作者：
FISCHER, A
影响因子：
20.3
作者：
Deregibus, Maria Chiara;Cantaluppi, Vincenzo;Camussi, Giovanni
通讯作者：
Camussi, Giovanni
DOI：
10.1016/0005-2795(71)90181-4
发表时间：
1971-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
CURTIS, J;BOURNE, FJ
通讯作者：
BOURNE, FJ
基于深度测序数据 isomiR 的 miRNA 基因簇和家族的全局表达分析
影响因子：
3.1
作者：
Guo, Li;Lu, Zuhong
通讯作者：
Lu, Zuhong