Identification and characterization of miRNAs expressed in the bovine ovary.
Identification and characterization of miRNAs expressed in the bovine ovary.
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DOI：
10.1186/1471-2164-10-443
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发表时间：
2009-09-18
期刊：
影响因子：
4.4
通讯作者：
Tesfaye D
中科院分区：
文献类型：
--
作者：
Hossain MM;Ghanem N;Hoelker M;Rings F;Phatsara C;Tholen E;Schellander K;Tesfaye D
MicroRNAs是一类主要的基因调控分子，在转录后水平上通过与靶mrna的序列互补发挥多种作用。这些mirna可以调节卵巢卵泡发生过程中多种基因的严格调控表达和相互作用。鉴定它们是了解mirna在不同生物学功能中引导的基因调控的第一步。尽管在不同物种和不同组织类型中鉴定miRNAs的努力越来越多，但对牛卵巢miRNAs知之甚少。本文报道了通过克隆、表达分析和靶标预测，鉴定和表征牛卵巢中表达的mirna。本研究构建的牛卵巢miRNA文库（5′独立连接克隆法）共克隆了50个已知miRNA和24个新miRNA。在所有鉴定的mirna中，有38个是牛的新mirna，它们来自43个具有独特二级结构的不同位点。虽然发现22个miRNAs前体位点在不止一个物种中保存良好，但发现16个是牛特异性的。大多数mirna被多次克隆，其中let-7a、let-7b、let-7c、miR-21、miR-23b、miR-24、miR-27a、miR-126和miR-143分别被克隆10次、28次、13次、4次、11次、7次、6次、4次和11次。对所有新mirna和部分加注释mirna在卵巢内不同结构和其他多个组织中的表达分析表明，一些mirna普遍存在，而另一些mirna在不同组织类型中表达差异。Bta-miR-29a定位于周期卵巢不同发育阶段的卵泡细胞中。mirna靶点的生物信息学预测、筛选和基因本体分析确定了卵巢功能的几个生物学过程和途径。本研究结果表明，mirna在牛卵巢中存在，从而阐明了它们在调节卵巢功能的多种分子和生理途径中的潜在作用。这一信息将使我们对牛卵巢miRNAs有更深入的了解，这些miRNAs可以进一步表征它们在卵泡发育和女性生育能力中的作用。
MicroRNAs are the major class of gene-regulating molecules playing diverse roles through sequence complementarity to target mRNAs at post-transcriptional level. Tightly regulated expression and interaction of a multitude of genes for ovarian folliculogenesis could be regulated by these miRNAs. Identification of them is the first step towards understanding miRNA-guided gene regulation in different biological functions. Despite increasing efforts in miRNAs identification across various species and diverse tissue types, little is known about bovine ovarian miRNAs. Here, we report the identification and characterization of miRNAs expressed in the bovine ovary through cloning, expression analysis and target prediction. The miRNA library (5'-independent ligation cloning method), which was constructed from bovine ovary in this study, revealed cloning of 50 known and 24 novel miRNAs. Among all identified miRNAs, 38 were found to be new for bovine and were derived from 43 distinct loci showing characteristic secondary structure. While 22 miRNAs precursor loci were found to be well conserved in more than one species, 16 were found to be bovine specific. Most of the miRNAs were cloned multiple times, in which let-7a, let-7b, let-7c, miR-21, miR-23b, miR-24, miR-27a, miR-126 and miR-143 were cloned 10, 28, 13, 4, 11, 7, 6, 4 and 11 times, respectively. Expression analysis of all new and some annotated miRNAs in different intra-ovarian structures and in other multiple tissues showed that some were present ubiquitously while others were differentially expressed among different tissue types. Bta-miR-29a was localized in the follicular cells at different developmental stages in the cyclic ovary. Bio-informatics prediction, screening and Gene Ontology analysis of miRNAs targets identified several biological processes and pathways underlying the ovarian function. Results of this study suggest the presence of miRNAs in the bovine ovary, thereby elucidate their potential role in regulating diverse molecular and physiological pathways underlying the ovarian functionality. This information will give insights into bovine ovarian miRNAs, which can be further characterized for their role in follicular development and female fertility as well.
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