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1.
本研究旨在发掘牛更多的microRNA(miRNA)信息及新的miRNA序列,为进一步研究miRNA的生物学功能奠定理论基础.运用高通量测序技术对来自一头西门塔尔公牛和一头荷斯坦母牛的多个组织的小RNA进行混池测序,对测序结果进行了生物信息学分析.随机选取了一条成熟miRNA:bta-miR-2346-5p和两条新预测的miRNAs:novel_miR-48和novel_miR-86,采用茎环RT-PCR进行验证.共鉴定了604条miRNAs,其中429条为牛的已知miRNAs,175条为新预测的miRNAs.通过茎环RT-PCR发现选取的这3条miRNAs在背最长肌、心肌、肝脏、脾脏、肺脏、肾脏、大肠、小肠中均有表达,证明了高通量测序结果的准确性.本试验得到的数据在一定程度上丰富了牛miRNA的数量和种类,为后续牛或其他物种的miRNA相关生物学研究提供了更加详实的信息. 相似文献
2.
《中国畜牧兽医》2015,(6)
本研究旨在发掘牛更多的microRNA(miRNA)信息及新的miRNA序列,为进一步研究miRNA的生物学功能奠定理论基础。运用高通量测序技术对来自一头西门塔尔公牛和一头荷斯坦母牛的多个组织的小RNA进行混池测序,对测序结果进行了生物信息学分析。随机选取了一条成熟miRNA:bta-miR-2346-5p和两条新预测的miRNAs:novel_miR-48和novel_miR-86,采用茎环RT-PCR进行验证。共鉴定了604条miRNAs,其中429条为牛的已知miRNAs,175条为新预测的miRNAs。通过茎环RT-PCR发现选取的这3条miRNAs在背最长肌、心肌、肝脏、脾脏、肺脏、肾脏、大肠、小肠中均有表达,证明了高通量测序结果的准确性。本试验得到的数据在一定程度上丰富了牛miRNA的数量和种类,为后续牛或其他物种的miRNA相关生物学研究提供了更加详实的信息。 相似文献
3.
《畜牧兽医学报》2015,(8)
本研究旨在通过比较基因组学和结构序列分析方法预测牛基因组中的新miRNAs并进行试验验证。根据miRNA分子序列具有一定保守性,将人、小鼠、绵羊、猪和狗5种哺乳动物已知的miRNA分子与NCBI中牛的全基因组序列(UMD3.1)对比,获得牛miRNAs;随机选取部分预测的miRNAs进行实时荧光定量PCR(qRTPCR)验证。结果,基于物种间序列同源比对共计预测到44条新的牛miRNAs,选取其中4条miRNAs,通过qRTPCR方法,发现他们在泌乳期中国荷斯坦牛乳腺、心、子宫和肝组织中均有表达。结果表明,基于比较基因组学和生物信息学预测新的miRNA分子可行,为奶牛基因表达调控及其性状形成机制研究提供了前期基础。 相似文献
4.
旨在发掘牛胎儿骨骼肌来源的成肌细胞分化过程中差异表达的miRNA,为进一步研究牛胎儿期成肌细胞分化的分子机理提供理论基础。本研究体外培养3头4月龄的牛胎儿骨骼肌来源的成肌细胞,运用高通量测序技术对成肌细胞分化起始和终末两个不同时期进行miRNA测序,并对测序结果进行生物信息学分析,鉴定差异表达的miRNA。随机选取差异表达miRNA,进行荧光定量PCR验证。结果表明,本研究从6个测序文库中共鉴定出已知miRNA 619个,其中差异表达miRNA 150个。差异表达miRNA中bta-miR-199a-5p等多个miRNA已经被证实与肌肉发育相关。通过靶基因预测富集到MAPK等多个与肌肉分化相关的通路。随机选取其中的8个miRNA进行荧光定量PCR验证,定量结果与测序结果完全一致。本研究鉴定出了150个与牛胎儿骨骼肌来源的成肌细胞分化相关的差异表达miRNAs。本研究结果将为研究miRNA调控牛胎儿骨骼肌发育提供科学依据。 相似文献
5.
微小RNA(miRNA)是一种内源性非编码小RNA,主要通过与m RNA上2~8位核苷酸碱基互补结合使目标m RNA失去稳定性来发挥作用。大量研究表明,miRNA在牛的脂肪沉积与代谢,脂肪细胞的生成、增殖与分化过程中起着至关重要的作用。为此,文章从miRNA的合成、mi R-378调控脂质代谢、mi R-27b等其他转录因子对脂质代谢的调控、miRNA调控脂质代谢通路和在牛其他组织中的调控作用等5方面综述了牛脂肪组织中miRNA的研究进展,并对其在牛的生长发育、肉品质改良、脂肪代谢调节和牛营养水平均衡等方面的研究应用进行了展望,以期为miRNA在牛脂肪组织中的深入研究提供一定参考。 相似文献
6.
根据miRNA进化的保守性,以已知动物的miRNA为搜索序列与山羊的UniGene进行BLAST比对,预测山羊新的miRNA,并通过RT-PCR和克隆测序进行验证。对新发现的山羊miRNA进行实时荧光定量,检测其在12个月皮肤及肌肉中的表达量;用基于UniGene的方法获得了5条山羊miRNA候选分子,通过RT-PCR和克隆测序验证,发现这5条miRNA分子在山羊的皮肤和肌肉中均有表达,其中由chi-miR-374b、chi-miR-421和chi-miR-421*构成了山羊miRNA基因簇,chi-miR-2284n是牛和山羊特异的。实时荧光定量结果显示,chi-miR-1839、chi-miR-374b和chi-miR-2284n在2月份皮肤中的表达量明显高于其他月份,chi-miR-421和chi-miR-421*在10月份皮肤中的表达量最高;发现了5条山羊新的miRNA(chi-miR-2284n、chi-miR-421*、chi-miR-421、chi-miR-1839和chi-miR-374b,登录号:JQ002550-JQ002554),补充了山羊miRNA数据库的不足。 相似文献
7.
《中国兽医杂志》2017,(10)
MicroRNA(miRNA)在炎症反应中起重要作用,本试验用miRNA测序技术研究了细菌脂多糖(Lipopolysaccharide,LPS)诱导的牛子宫内膜细胞miRNA差异表达。用1μg/mL的LPS处理牛子宫内膜细胞24 h,测定细胞上清液IL-6和IL-8分泌量,对细胞进行miRNA测序,并用荧光定量PCR验证测序结果。结果表明,LPS可诱导牛子宫内膜细胞11个miRNA表达上调、9个miRNA表达下调。差异表达miRNA的靶基因注释到生物过程的GO term共20个,注释到细胞组分的GO term共12个,注释到分子功能的GO term共20个;差异表达miRNA靶基因主要富集于PI3K-Akt和MAPK等信号通路。结果提示,miRNA在LPS诱导的牛子宫内膜细胞炎症反应中起重要作用,其作用与PI3K-Akt和MAPK等信号通路激活有关。 相似文献
8.
近年来,miRNA已成为生命科学领域的研究热点。文章综述了miRNA在真核和原核生物中的功能,以及miRNA的分子克隆、生物信息学等预测和鉴定方法,并阐述了miRNA研究中的问题及应用前景。 相似文献
9.
本试验旨在构建乾华肉用美利奴羊次级毛囊不同发育时期的小RNA文库和预测筛选miRNA的候选靶基因。通过高通量测序技术构建乾华肉用美利奴羊次级毛囊小RNA文库|采用荧光定量PCR对高通量测序结果进行验证|利用miRDeep2软件对miRNA（长度为20~24 nt的小RNA）进行鉴定|利用Targetscan v7.0、RNA22 v2.0和MiRanda三个在线靶基因预测软件对miR-1-3p的靶基因进行预测分析。成功获得乾华肉用美利奴羊次级毛囊生长期、退行期、休止期小RNA纯净reads分别为10279515、11712215和10926258。分别基于荧光定量与高通量测序的5个miRNA的相对表达量结果趋势基本一致。从次级毛囊生长期、退行期和休止期分别鉴定获得miRNA 1250、1304个和1266个。确定成纤维细胞生长因子14（FGF14）和类胰岛素生长因子Ⅰ 受体（IGF1R）为miR-1-3p的候选靶基因。研究结果为后续靶基因验证及确定调控乾华肉用美利奴羊毛囊周期性发育的候选基因奠定基础。
[关键词] 乾华肉用美利奴羊|小RNA文库|miR-1-3p|靶基因预测 相似文献
11.
Peter Vegh Amir B.K. Foroushani David A. Magee Matthew S. McCabe John A. Browne Nicolas C. Nalpas Kevin M. Conlon Stephen V. Gordon Daniel G. Bradley David E. MacHugh David J. Lynn 《Veterinary immunology and immunopathology》2013
MicroRNAs (miRNAs) are important regulators of gene expression and are known to play a key role in regulating both adaptive and innate immunity. Bovine alveolar macrophages (BAMs) help maintain lung homeostasis and constitute the front line of host defense against several infectious respiratory diseases, such as bovine tuberculosis. Little is known, however, about the role miRNAs play in these cells. In this study, we used a high-throughput sequencing approach, RNA-seq, to determine the expression levels of known and novel miRNAs in unchallenged BAMs isolated from lung lavages of eight different healthy Holstein–Friesian male calves. Approximately 80 million sequence reads were generated from eight BAM miRNA Illumina sequencing libraries, and 80 miRNAs were identified as being expressed in BAMs at a threshold of at least 100 reads per million (RPM). The expression levels of miRNAs varied over a large dynamic range, with a few miRNAs expressed at very high levels (up to 800,000 RPM), and the majority lowly expressed. Notably, many of the most highly expressed miRNAs in BAMs have known roles in regulating immunity in other species (e.g. bta-let-7i, bta-miR-21, bta-miR-27, bta-miR-99b, bta-miR-146, bta-miR-147, bta-miR-155 and bta-miR-223). The most highly expressed miRNA in BAMs was miR-21, which has been shown to regulate the expression of antimicrobial peptides in Mycobacterium leprae-infected human monocytes. Furthermore, the predicted target genes of BAM-expressed miRNAs were found to be statistically enriched for roles in innate immunity. In addition to profiling the expression of known miRNAs, the RNA-seq data was also analysed to identify potentially novel bovine miRNAs. One putatively novel bovine miRNA was identified. To the best of our knowledge, this is the first RNA-seq study to profile miRNA expression in BAMs and provides an important reference dataset for investigating the regulatory roles miRNAs play in this important immune cell type. 相似文献
12.
【目的】 探索胚胎移植前供体牛与受体牛血浆外泌体miRNA的表达差异, 以明确血浆外泌体miRNA在牛早期妊娠中的作用及其调控机制。【方法】 以3~6岁、体重480~600 kg的夏南牛作为研究对象, 选取10头供体牛进行同期发情、超数排卵和人工授精, 23头受体牛只做同期发情处理。在人工授精后第7天, 冲洗供体牛子宫以获取囊胚, 选取3头囊胚数相近的供体牛及3头与供体牛体重和年龄均相近的受体牛, 颈静脉采血, 进行血浆外泌体的分离与鉴定; 然后提取血浆外泌体miRNA, 并检测其表达量; 采用R语言中的DESeq差异算法计算P值, 并筛选出P<0.05的miRNA, 对差异表达的miRNA进行靶基因预测、GO功能富集分析和KEGG信号通路分析。【结果】 6个样本的囊泡粒径均在135 nm左右, 符合外泌体的特征。与供体牛相比, 受体牛中有9个miRNAs表达显著上调(P<0.05), 13个miRNAs显著下调(P<0.05);22个差异表达的miRNAs中, 有15个miRNAs预测出无重复的靶基因2 990个。GO功能富集分析和KEGG信号通路分析的结果表明, 这些靶基因主要富集在与生物黏附(biological adhesion)、定位(localization)、细胞连接(cell junction)功能有关的通路上, 显著富集的信号通路与黏着斑(focal adhesion)、黏着连接(adherens junction)有关, 提示血浆外泌体miRNA可能参与调控胚胎着床。【结论】 研究结果可为筛选和探究影响胚胎着床的血浆外泌体miRNA提供参考, 并为进一步阐明血浆外泌体miRNA在母牛早期妊娠调控中的作用提供依据。 相似文献
13.
The ovarian follicle components must provide an ideal environment to ensure the success of reproductive processes, and communication between follicular cells is crucial to support proper oocyte growth. Recently, it has been demonstrated that the presence of extracellular vesicles (EVs) carrying microRNAs (miRNAs) in follicular fluid represents an important autocrine and paracrine communication mechanism inside the ovarian follicle. In this study, we tested the hypothesis that the miRNA content of EVs isolated from ovarian follicular (granulosa and cumulus–oocyte complexes) cell‐conditioned culture media is dependent upon cell type. We initially screened bovine granulosa cells (GCs) and cumulus–oocyte complexes (COCs), as well as their derived EVs for 348 miRNAs using real‐time PCR, and detected 326 miRNAs in GCs and COCs cells and 62 miRNAs in GCs and COCs EVs. A bioinformatics analysis of the identified cell‐specific and differentially expressed miRNAs predicted that they likely modulate important cellular processes, including signalling pathways such as the PI3K‐Akt, MAPK and Wnt pathways. By investigating the origins of miRNAs within the follicular fluid, the results of this study provide novel insights into follicular miRNA content and intercellular communication that may be of invaluable use in the context of reproductive technologies, diagnostic of ovarian‐related diseases and/or the identification of biomarkers for oocyte and embryo quality. 相似文献
14.
MicroRNAs (miRNAs) represent a newly identified class of non‐protein‐coding ～22 nt small RNAs which play important roles in multiple biological processes by degrading targeted mRNAs or repressing mRNA translation. Here we present an expressed sequence tag (EST)‐based combined approach for the detection of novel porcine miRNAs. This was initiated by using previously known miRNA sequences from Homo sapiens (human) and Mus musculus (mouse) to blast the databases of Sus scrofa (pig) EST. A total of 65 new miRNAs were detected following a range of filtering criteria. Using these new potential miRNA sequences, we further obtained the publicly available porcine mRNA database from NCBI and detected 48 586 potential target hits using a software RNA hybrid. So far, compared to human and mouse, fewer miRNAs (only 54 miRNAs) were identified in Sus scrofa species. These 65 new miRNAs and their targets in pig have been run through miRHelper to yield data that may help us better understand the possible role of miRNAs in regulating the growth and development of pigs. These findings suggest that EST analysis is a good alternative strategy for identifying new miRNA candidates, their targets and other genes. 相似文献
15.
MicroRNA（miRNA）是一类约22个核苷酸组成的单链非编码RNA,在分化、发育、肿瘤形成等方面起着重要作用。本研究对弓形虫RH株感染小鼠脾细胞miRNA的表达进行了特异性基因芯片检测分析,并应用荧光定量RT-PCR方法进行验证。结果表明,感染鼠脾细胞中与免疫应答及细胞增殖和肿瘤发生相关的三大类miRNA中,有39种表达下调,同时有36种表达上调。上述结果揭示,弓形虫感染机体后伴随着靶细胞功能性miRNA表达谱的显著改变,这为进一步研究弓形虫感染致病的分子机制开辟了新的方向。 相似文献
16.
肌内脂肪（intramuscular fat,IMF）含量是评判猪肉品质的关键指标,其可影响猪肉的嫩度、剪切力、多汁性和风味等。微小RNA （microRNA,miRNA）是一类长度约为22 nt的短链非编码RNA,在胚胎发育、成脂分化、肌纤维形成、神经调节、免疫应答等多种生理过程中发挥重要作用。越来越多的研究表明,miRNA在猪肌内脂肪沉积过程中发挥重要调控作用,是脂质代谢的重要调控因子。作者通过归纳国内外关于miRNA调控猪肌内脂肪沉积的相关研究发现,miR-34a、miR-125a-5p、miR-32-5p等通过靶向作用转录因子Krüppel样因子（Krüppel-like factors,KLF）家族成员调控猪肌内脂肪沉积,miR-130a通过靶向作用过氧化物酶体增殖物激活受体（peroxisome proliferator activated receptor,PPAR）家族成员调控猪肌内脂肪沉积,miR-34a、miR-17-5p和miR-125a-5p等通过靶向作用其他家族成员,如长链酯酰辅酶A合成酶4（acyl-CoA synthetase long chain family member 4,ACSL4）、核受体共激活因子3（nuclear receptor coactivator 3,NCOA3）等来调控猪肌内脂肪沉积。然而miRNA调控猪肌内脂肪的具体作用机制尚不完全清楚,还需要进一步探究。作者通过梳理目前已经证实的与猪肌内脂肪沉积相关的miRNA,整理相关miRNA的靶基因以及主要作用通路,以期为筛选肌内脂肪相关miRNA提供参考,为改善肉质提供新的思路和策略,为阐明miRNA在猪脂质代谢中的作用机制提供参考。 相似文献
17.
Lucia Unger Vinzenz Gerber Alicja Pacholewska Tosso Leeb Vidhya Jagannathan 《Veterinary and comparative oncology》2019,17(1):107-117
Serum and whole blood microRNA (miRNA) fingerprints have been proposed as a new class of non‐invasive human cancer biomarkers. In this study, we compared equine sarcoid (ES) disease‐specific serum and whole blood miRNA fingerprints and correlated them to miRNA expression in sarcoid tissue. After high throughput sequencing, miRNA differential expression analysis between six ES‐affected and five control horses was carried out in serum and whole blood using a DESeq algorithm, accounting for the influence of hemolysis and the white blood cell count. Target gene, pathway prediction and enrichment analyses were conducted using TarBase, mirPath and GeneCodis. After exclusion of 4 hemolyzed out of a total of 11 serum samples, 9 miRNAs were found to be differentially expressed in serum of ES vs control horses. In whole blood, all 11 samples showed normal white blood cell counts and 19 miRNAs were found to be differentially expressed. A total of 2/9 serum and 7/19 whole blood differentially expressed miRNAs were also highly expressed at the tissue level and their predicted target genes were associated with cancer pathways. Serum and whole blood miRNA expression allowed discrimination between ES and control horses and merits further validation in a larger study cohort. The use of whole blood might be superior because it has higher miRNA content and is less influenced by pre‐analytical variables compared to serum. Concurrent dysregulation of single miRNAs in tissue and blood suggests a possible biological function of circulating miRNAs. 相似文献
18.
脂肪组织是动物机体重要的能量代谢及内分泌器官,选择性的脂肪沉积对动物肉类的感官品质、风味性和加工特性具有至关重要的作用,因此动物不同部位脂肪沉积的特异性调控因子及其作用分子机理备受研究者的关注。microRNA（miRNA）是一类长度为22 nt左右的非编码小RNA,近年来采用组学技术对具有表型差异的脂肪组织和脂肪细胞进行高通量测序,筛选发现了许多差异表达的miRNAs,这些miRNAs可通过与靶基因mRNA相结合发挥生物学功能,对不同部位脂肪沉积调控具有重要作用。鉴于此,本文将从miRNA在动物皮下脂肪组织和肌内脂肪的调控作用等方面进行综述,为后续研究miRNA调控动物脂肪组织沉积的作用及机制提供理论参考和新的思路。 相似文献
19.
Dilda F Gioia G Pisani L Restelli L Lecchi C Albonico F Bronzo V Mortarino M Ceciliani F 《Veterinary journal (London, England : 1997)》2012,192(3):514-516
MicroRNAs (miRNAs) are a family of regulatory molecules involved in many physiological processes, including activation of cells of the immune system. This study investigated the effect of Escherichia coli lipopolysaccharide (LPS) and Staphylococcus aureus enterotoxin B (SEB) on the expression of five miRNAs involved in the inflammatory response, including miR-9, miR-125 b, miR-155, miR-146 a and miR-223, in bovine CD14(+) cells (monocytes). Incubation of monocytes with SEB induced down-regulation of miR-155, miR-223 and miR-125 b, but not the anti-inflammatory miRNA miR-146 a. Conversely, incubation with LPS upregulated both miR-155 and miR-146 a. In vitro incubation of isolated CD14(+) bovine monocytes with LPS and SEB elicited different and opposite expression of miRNAs reportedly involved in inflammatory reactions. 相似文献
20.
Z. Wicik M. Gajewska A. Majewska D. Walkiewicz E. Osińska T. Motyl 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2016,133(1):31-42
MicroRNAs (miRNAs) are small non‐coding RNAs that participate in the regulation of gene expression. Their role during mammary gland development is still largely unknown. In this study, we performed a microarray analysis to identify miRNAs associated with high mammogenic potential of the bovine mammary gland. We identified 54 significantly differentially expressed miRNAs between the mammary tissue of dairy (Holstein‐Friesian, HF) and beef (Limousin, LM) postpubertal heifers. Fifty‐two miRNAs had higher expression in the mammary tissue of LM heifers. The expression of the top candidate miRNAs (bta‐miR‐10b, bta‐miR‐29b, bta‐miR‐101, bta‐miR‐375, bta‐miR‐2285t, bta‐miR‐146b, bta‐let7b, bta‐miR‐107, bta‐miR‐1434‐3p) identified in the microarray experiment was additionally evaluated by qPCR. Enrichment analyses for targeted genes revealed that the major differences between miRNA expression in the mammary gland of HF versus LM were associated with the regulation of signalling pathways that are crucial for mammary gland development, such as TGF‐beta, insulin, WNT and inflammatory pathways. Moreover, a number of genes potentially targeted by significantly differentially expressed miRNAs were associated with the activity of mammary stem cells. These data indicate that the high developmental potential of the mammary gland in dairy cattle, leading to high milk productivity, depends also on a specific miRNA expression pattern. 相似文献