Identification of Differentially Expressed miRNAs and Their Potential Targets During Fish Ovarian Development
Identification of Differentially Expressed miRNAs and Their Potential Targets During Fish Ovarian Development
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DOI：
10.1095/biolreprod.112.105361
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发表时间：
2013-05-01
影响因子：
3.6
通讯作者：
Bobe, Julien
中科院分区：
文献类型：
--
作者：
Juanchich, Amelie;Le Cam, Aurelie;Bobe, Julien
卵子发生是一个复杂的过程，需要特定基因的协调有序表达，最终导致雌性配子从卵巢中释放出来。在本研究中，我们旨在研究mirna在硬骨鱼中对这一关键生物学过程的调节作用，使用一个同时生长的卵母细胞发育的模型。利用mirna在系统发育上较远的物种之间的强序列保守性，我们设计了一个通用的微阵列，显示大多数已知的脊索动物mirna。这使我们首次在任何脊椎动物物种中提供了卵子发生过程中卵巢miRNome的概述。我们鉴定了13个差异表达的miRNA，并且在卵发生的每个步骤中观察到至少一个miRNA的差异表达。在卵发生的几个阶段观察到几种mirna的高差异表达，随后用定量PCR证实。通过利用在同一样本集中获得的基因表达数据来完善靶基因的计算机预测，我们提供了强有力的证据，证明miRNAs靶向卵子发生的主要参与者，包括参与类固醇形成速率限制步骤的基因、参与卵母细胞发育促性腺激素控制的基因，以及参与排卵、卵母细胞水合作用和获得卵母细胞受精后支持进一步发育的能力的基因。卵母细胞发育能力)。总之，这些观察结果强调了mirna在卵子发生过程中对雌性配子形成的调节和成功的重要性。
Oogenesis is a complex process requiring the coordinated sequential expression of specific genes and ultimately leading to the release of the female gamete from the ovary. In the present study we aimed to investigate the contribution of miRNAs to the regulation of this key biological process in teleosts using a model in which growing oocytes develop simultaneously. Taking advantage of the strong sequence conservation of miRNAs among phylogenetically distant species, we designed a generic microarray displaying most known chordate miRNAs. It allowed us to provide an overview of the ovarian miRNome during oogenesis for the first time in any vertebrate species. We identified 13 differentially expressed miRNAs, and a differential expression of at least one miRNA was observed at each step of oogenesis. A surprisingly high differential expression of several miRNAs was observed at several stages of oogenesis and subsequently confirmed using quantitative PCR. By refining in silico prediction of target genes with gene expression data obtained within the same sample set, we provide strong evidence that miRNAs target major players of oogenesis, including genes involved in rate-limiting steps of steroidogenesis and those involved in gonadotropic control of oocyte development, as well as genes involved in ovulation, oocyte hydration, and acquisition of the ability of the oocyte to support further development once fertilized (i.e., oocyte developmental competence). Together, these observations stress the importance of miRNAs in the regulation and success of female gamete formation during oogenesis.