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虹鳟鱼排卵后老化导致卵子退化相关的线粒体基因组编码小RNA的鉴定

Identification of Mitochondrial Genome-Encoded Small RNAs Related to Egg Deterioration Caused by Postovulatory Aging in Rainbow Trout.

作者信息

Ma Hao, Weber Gregory M, Wei Hairong, Yao Jianbo

机构信息

National Center for Cool and Cold Water Aquaculture, Kearneysville, WV, 25430, USA.

Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV, 26506, USA.

出版信息

Mar Biotechnol (NY). 2016 Oct;18(5):584-597. doi: 10.1007/s10126-016-9719-3. Epub 2016 Oct 24.

Abstract

Many factors have been reported to affect rainbow trout egg quality, among which, postovulatory aging is one of the most significant causes as reared rainbow trout do not usually volitionally oviposit the ovulated eggs. In order to uncover the genetic regulation underling egg deterioration caused by postovulatory aging in rainbow trout, mitochondrial genome-encoded small RNA (mitosRNAs) were analyzed from unfertilized eggs on Days 1, 7, and 14 postovulation with fertilization rates of 91.8, 73.4, and less than 50 %, respectively. A total of 248 mitosRNAs were identified from Illumina high-throughput sequencing of the small RNA libraries derived from the eggs of ten females. Ninety-eight of the small RNAs exhibited more than a threefold difference in expression between eggs from females exhibiting high fertilization rates at Day 1 and low fertilization rates at Day 14. The differentially expressed mitosRNAs were predominantly derived from mitochondrial D-loop, tRNA, rRNA, COII, and Cytb gene regions. Real-time quantitative PCR analysis was carried out for 14 differentially expressed mitosRNAs, of which, 12 were confirmed to be consistent with the sequencing reads. Further characterization of the differentially expressed mitosRNAs may lead to the development of new biomarkers for egg quality in rainbow trout.

摘要

据报道,许多因素会影响虹鳟鱼卵的质量,其中,排卵后老化是最重要的原因之一,因为养殖的虹鳟鱼通常不会主动产出已排卵的卵。为了揭示虹鳟鱼排卵后老化导致卵质量下降的遗传调控机制,对排卵后第1天、第7天和第14天未受精卵的线粒体基因组编码小RNA(mitosRNAs)进行了分析,受精率分别为91.8%、73.4%和低于50%。通过对来自10只雌性虹鳟鱼卵的小RNA文库进行Illumina高通量测序,共鉴定出248种mitosRNAs。其中98种小RNA在第1天受精率高的雌性虹鳟鱼卵和第14天受精率低的雌性虹鳟鱼卵之间表现出超过三倍的表达差异。差异表达的mitosRNAs主要来源于线粒体D环、tRNA、rRNA、COII和Cytb基因区域。对14种差异表达的mitosRNAs进行了实时定量PCR分析,其中12种被证实与测序读数一致。对差异表达的mitosRNAs进行进一步表征可能会导致开发虹鳟鱼卵质量的新生物标志物。

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