College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China.
Hubei Hongshan Laboratory, Wuhan, 430070, China.
BMC Genomics. 2023 Mar 16;24(1):121. doi: 10.1186/s12864-023-09220-9.
Egg quality is a major concern in fish reproduction and development. An effective evaluation of egg quality prior to fertilization is helpful in improving the fertilization rate and survival rate of the larva. In this study, we aim to identify quality instructors from the combination study of fertilization rate, hatching rate, embryo malformation rate and gene expression profile.
Eggs from 25 female fish were fertilized with sperm from the same fish. The egg quality was determined by the fertilization rates, hatching rate and embryo malformation rate and divided into three categories, low-quality (< 35%), medium-quality (35 to 75%), and high-quality (> 75%). Due to the distinct difference in fertilization, hatching and embryo malformation rate between low-quality eggs and high-quality eggs, these two groups were considered for the identification of quality markers. Then RNA-seq was performed for the originally preserved eggs from the low-quality group and high-quality group. We profiled the differentially expressed genes and identified a group of RNA-binding proteins (RBPs) as potential regulators. Gene function analysis indicated that most of these genes were enriched in RNA-regulated pathways including RNA processing. The RBPs were more related to egg quality from the PLS-DA analysis. Finally, gene expression was validated by qRT-PCR.
We found a cluster of RBP genes including igf2bp3, zar1, elavl1, rbm25b and related regulatory factors including yy1, sirt1, anp32e, btg4 as novel biomarkers of egg quality.
卵子质量是鱼类繁殖和发育的主要关注点。在受精前对卵子质量进行有效评估有助于提高受精率和幼虫成活率。在这项研究中,我们旨在通过受精率、孵化率、胚胎畸形率和基因表达谱的综合研究来确定质量指标。
从 25 条雌鱼中取出的卵子与同一条雄鱼的精子受精。通过受精率、孵化率和胚胎畸形率来确定卵子质量,并将其分为三个类别:低质量(<35%)、中质量(35%至 75%)和高质量(>75%)。由于低质量卵子和高质量卵子在受精、孵化和胚胎畸形率方面存在明显差异,因此将这两组用于鉴定质量标志物。然后对低质量组和高质量组原保存的卵子进行 RNA-seq 分析。我们对差异表达基因进行了分析,并鉴定了一组 RNA 结合蛋白(RBPs)作为潜在的调控因子。基因功能分析表明,这些基因大多数富集在 RNA 调控途径中,包括 RNA 加工。PLS-DA 分析表明,RBPs 与卵子质量的相关性更强。最后,通过 qRT-PCR 验证了基因表达。
我们发现了一组 RBP 基因,包括 igf2bp3、zar1、elavl1、rbm25b 和相关调节因子 yy1、sirt1、anp32e、btg4,它们可能是卵子质量的新标志物。