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鉴定和差异表达微 RNA 在睾丸和卵巢的史氏鲟 (Acipenser schrenckii)。

Identification and differential expression of microRNAs in testis and ovary of Amur sturgeon (Acipenser schrenckii).

机构信息

Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Institute of Applied Biological Resources, Guangzhou, Guangdong, China.

出版信息

Gene. 2018 Jun 5;658:36-46. doi: 10.1016/j.gene.2018.03.014. Epub 2018 Mar 7.

DOI:10.1016/j.gene.2018.03.014

PMID:29524574

Abstract

BACKGROUND

MicroRNAs (miRNAs) cooperate with sex-related genes in post-transcriptional regulation and play extremely important roles in the establishment of sexually dimorphic traits in animals. However, the gonad miRNAs and expression patterns of miRNAs in sturgeon have not been investigated.

METHODS

In the present study, we used high-throughput small RNA sequencing (RNA-Seq) to discover gonad miRNAs from the ovaries and testes of Amur sturgeons (Acipenser schrenckii). Further, microarray and real-time PCR assays were performed to identify the expression patterns of gonad miRNAs.

RESULTS

As a result, a total of 679 conserved and 51 novel miRNAs were successfully discovered in the gonads of A. schrenckii. Moreover, we found wide sequence variations (isomiRs) in gonad miRNAs, including 5' and 3' isomiRs. Our microarray analysis further characterized the 730 miRNAs expression profiles, which indicated that 117 differentially expressed miRNAs were detected with sex-biased patterns: 71 testis-biased and 46 ovary-biased miRNAs. Based on bioinformatics prediction, we found that there were functional differences between the testis-biased and ovary-biased miRNA targets involved in reproductive-related GO and KEGG pathways. Further, the association of the differentially expressed miRNAs and sex-related target mRNAs was uncovered. Finally, the expression patterns of 11 sex-biased miRNAs and 7 sex-related targets were validated in testes and ovaries using real-time PCR. Putative, negatively expressed miRNA-mRNA relationships were confirmed, such as Dmrt1 and asc-miR-2779, AR and asc-miR-203b-3p, foxl2 and asc-miR-30d.

CONCLUSION

This study provides information regarding the gonad miRNAs in sturgeon. The differential expression miRNAs in the gonads will help us to further understand the role of miRNA-mediated post-transcriptional regulation in the ovary and testis of sturgeon.

摘要

背景

微小 RNA（miRNAs）与性别相关基因在转录后调控中合作，在动物性别二态性特征的建立中发挥着极其重要的作用。然而，鲟鱼的性腺 miRNA 及其 miRNA 的表达模式尚未得到研究。

方法

本研究采用高通量小 RNA 测序（RNA-Seq）技术从史氏鲟（Acipenser schrenckii）的卵巢和精巢中发现性腺 miRNA。进一步通过微阵列和实时 PCR 检测来鉴定性腺 miRNA 的表达模式。

结果

结果，在史氏鲟的性腺中共成功发现了 679 个保守和 51 个新的 miRNA。此外，我们发现了性腺 miRNA 中广泛的序列变异（isomiRs），包括 5' 和 3' isomiRs。我们的微阵列分析进一步表征了 730 个 miRNA 表达谱，结果表明，检测到 117 个具有性别偏倚模式的差异表达 miRNA：71 个睾丸偏倚和 46 个卵巢偏倚 miRNA。基于生物信息学预测，我们发现睾丸偏倚和卵巢偏倚 miRNA 靶标在生殖相关 GO 和 KEGG 途径中的功能存在差异。进一步揭示了差异表达 miRNA 与性别相关靶 mRNA 的关联。最后，使用实时 PCR 在睾丸和卵巢中验证了 11 个性别偏倚 miRNA 和 7 个性别相关靶基因的表达模式。确认了推定的负表达 miRNA-mRNA 关系，如 Dmrt1 和 asc-miR-2779、AR 和 asc-miR-203b-3p、foxl2 和 asc-miR-30d。